Expression and Prognostic Significance of Stem Cell Marker CD133 in Survival Rate of Colon Cancer Patients

Expression and Prognostic Significance of Stem Cell Marker CD133 in Survival Rate of Colon Cancer Patients | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Expression and Prognostic Significance of Stem Cell Marker CD133 in Survival Rate of Colon Cancer Patients Mohammad Tolooei, Hassan Ehteram, Hamed Haddad Kashani This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-1538147/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: Today, colon cancer is one of the most common types of gastrointestinal cancers CD133 as a known cancer stem cell marker has been found effective in cell proliferation and differentiation in various cancers, including colon. We aimed to investigate the relationship between the CD133 expression in colon cancer with prognostic factors and the survival rate of colon cancer patients by immunohistochemistry. Materials and Methods: Formalin- fixed-paraffinembedded (FFPE) tissues was taken from colon patients. Histopathology examination was done using hematoxylin and eosin staining. Immunohistochemistry was performed to determine CD133 expression. An association between CD133 expression and clinic pathological profile was then analyzed. Results: There was a statistically significant difference between CD133 protein expression and sex, cancer stage and lymphatic invasion (p=0.044, p=0.131 and p=0.002, respectively). However, no significant correlation was identified between the CD133 expression and other factors such as: age of CRC patients ( p -value=0.267), tumor location ( p -value=0.494), tumor differentiation grade ( p -value=0.263), neural tissue invasion and 5 years’ survival (p value=0.054). Conclusion: CD133 is a useful predictive or prognostic biomarker for CRC in clinical assessment and may serve as a potential therapeutic target for CRC. Carcinoma Colon Immunohistochemistry Neoplastic stem cells Rectum Introduction Colon cancer is one of the most prevalent cancers in the world and a major cause of cancer-related mortality [ 1 – 3 ]. In Iran, colon cancer is the third most common cancer in men and fourth common cancer in women [ 4 ]. It is suggested that the highly tumorigenic, chemotherapy resistant affect survival rate in colon cancer [ 5 – 8 ]. Targeted therapies have promising potential improvement of survival in colon cancer patients [ 9 ]. Recently, cancer stem cells (CSCs) have been known due to their ability of self-renew, self-repair and their role in the tumor initiation, metastasis, therapeutic resistance and recurrence [ 10 – 12 ]. Different markers have been also found to be expressed on the surface of CSCs among which CD133 has been paid the special attention. The CD133/prominin-1 is a 120-kDa 5-transmembrane domain antigen on chromosome 4p15.32 and mainly localized in membrane protrusions [ 13 , 14 ]. CD133 first discovered in hematopoietic stem and progenitor cells by Yin et al [ 15 ]. Previous studies identified CD133 as a putative CSC marker in cancer cells [ 16 – 19 ] with ability to initiate tumor growth [ 20 – 24 ]. This CSC marker now widely used to identify cancer stem cells in many cancers such as brain, colon, and lung cancers [ 25 – 28 ]. CD133 is normally expressed by primitive hematopoietic stem cells from adult blood and bone narrow, human fetal liver and stem cells in cord blood and peripheral blood, endothelial progenitor cells, epithelial cells and neural and glial stem cells, [ 29 – 31 ]. Several studies have suggested that high CD133 expression is associated with the clinic-pathologic characteristics such as poor prognosis, distant metastasis, survival rate and even chemotherapy resistance in colon cancer patients So, as a CRC-initiating CSC marker, CD133 expression can be used for developing a novel therapeutic approaches in colon cancer [ 32 – 40 ]. In this study, we investigated the association between the expression and prognostic significance of the CSC marker CD133 with clinic-pathologic features of CRC patients including survival rate, using the immunohistochemistry (IHC) method. Methods And Materials Tissue samples In this study, 34 formalin- fixed-paraffinembedded (FFPE) tumor tissues were selected from 34 patients underwent surgical resection from 2008–2019 in Shahid Beheshti Hospital in Kashan, Iran. Written consent was obtained from all subjects and the study was approved by the Research Ethical Committee of Kashan University of Medical Sciences, Iran. All clinic-pathologic data were verified using the hematoxylin and eosin (H&E) stained pathologic slides, patients’ medical files and pathology record. Tumors were classified according to the American Joint Committee on Cancer (AJCC) and the International Union Against Cancer (UICC) TNM criteria [93]. Clinic-pathologic parameters were included age, gender, tumor location, tumor stage, tumor differentiation grade, lymphatic and neural tissue invasion, 5 years’ survival and tumor recurrence. A colon cancer with detected positive immunostaining for CD133 considered as a positive control. Cases and controls were examined by two expert pathologists. Positive reaction was defined by the presence of at least 10% of cancer cells displaying either membranous and/or cytoplasmic staining. A number of tumor samples were excluded from the study due to inappropriate paraffin blocks, low quality staining and unavailable data. Also, the small size specimens prepared by biopsy which was inadequate for performing histopathological examinations were excluded. CD133 Immuno-histochemical staining The expression level of CD133 was determined in CRC sections by the Immunohistochemical staining (IHC) method as follows: Formalin-fixed, paraffin-embedded CRC tissue blocks were sectioned at 5-µm thickness for IHC staining. All slides were deparaffinized with xylene, and rehydrated in a graded ethylic alcohol (50%, 70%, 80% and 100%) followed by distilled water. After washing 3 times with phosphate-buffered saline (PBS) buffer every 5 minutes, endogenous peroxidase was inactive by immersing the sections in 1-0.1% H 2 O 2 for 5min. Then, for heat-induced antigen unmasking, sample tissues were treated in sodium citrate buffer (pH:6.0) at 95°C for 5 minutes in a microwave oven and allowed to cool for 1 hour at room temperature. CD133 expression was identified and visualized after reaction with primary Anti CD133 antibody (1:100) at 4˚C for one night. After washing the slides three times with PBS, a secondary antibody (peroxides- conjugated) was used to detect the primary antibody, and slides were incubated for 30 minutes in room temperature. Again, after three times washing in PBS for every 5 minutes, samples were incubated with 3, 3´-diaminobenzidine tetrahydrochloride-chromogen (DAB) for 1–5 minutes for visualizing immunostaining and finally counterstained with hematoxylin dye and dehydrated in ethylic alcohol serial dilution (60%,80% and 100%) and cleared in xylene (100%). Our negative control was the tissue that was covered with just buffer without the primary antibody. Prepared specimens were studied using light microscope. Imaging analysis of immunohistochemical staining of the specimens was evaluated by one pathologist without knowledge about the patients' clinic-pathological characteristics. The CD133 expression was assessed by observing the appearance of yellow to brown color in the cytoplasmic and membrane of glandular epithelial cells. A score of less than 10% CD133 cells (low CD133 expression) considered as negative and more than 10% CD133 cells (high CD133 expression) as positive, respectively. The staining intensity was evaluated according a ranging from non-stained (negative) to strong (positive): 0; non-staining, + 1; weakly staining, + 2; moderate staining, + 3; strong staining. Statistical analysis All statistical analyses were performed using SPSS software version 20 (SPSS Inc., Chicago, IL., USA). The results were expressed as the means ± standard deviation. The statistical significance of differences between immunohistochemical CD133 expression levels and clinicopathological features of CRC patients were evaluated using Chi-square and one-way ANOVA, or Mann-Whitney test or Kruskal-Wallis parametric bread tests (based on the results of Kolmogorov-Smirnov test), as appropriate. Survival was evaluated using the Kaplan-Meier method with a log-rank test. p -values of < 0.05 were considered statistically significant. Results This study included 34 tissue samples from 15 (44.1) male patients and 19 (55.9%) female patients with a median age of 62 (± 15.35) years at the time of diagnosis. The distribution of CRC patients with colon cancer according to demographic features is presented in Table 1 . Table 1 Characteristics of 38 CRC patients and their prognostic factors. Variable ( No.%) Gender Male 15 (44.1) Female 19 ( 55.9) Age (mean) (± SD) (years) 62 (± 15.35) Tumor location Ascendance colon 14 (429.4%) Transverse 1(3.0%) Sekom 6 (18.2%) Sigmoid 8 (2.2%) 8(24.2%) Descending 1 (3.0%) Rectum colon 2 (6.1%) Dariche ileocecal 2(6.1%) Histopathology Grade Well-differentiated ( G1) 21 (61.8%) Moderately differentiated (G2) 11 (32.4%) Poorly differentiated (G3) 2(5.8%) CD133 expression High Level 12 ( 35.3%) Low Level 9 (26.5%) 5 years’ survival Positive 26 (76.5%) Negative 8 ( 23.5%) Lymphatic invasion Positive 20 ( 58.8%) Negative 14 (41.2%0 Tumor stage 1 9 (26.5%) 2a 7 ( 20.6) 2b 4 (11.8%) 3a 1 (2.9%) 3b 8 (23.5%) 3c 5 (14.7%) Neuronal invasion Positive 4 (11.8%) Negative 30 (88.2%) The pathological results showed (as shown at Table 1 ) that the most tumors were located in ascendance colon 13 (39.4%) and in Sigmoid 8 (2.2%), while lowest tumor location was descending 1 (3.0%) and transverse 1 (3.0%). Also based on tumor differentiation degree, there were 21 (61.8%) well-differentiated (G1), 11 (32.4%) moderated differentiated (G2) and 2(5.8%) poor differentiation (G3) tumors. The highest prevalence of tumor stages was stage I (26.5%) and stage 3b (23.5%), while the lowest frequency was observed at stage 3a (2.9%). Moreover, lymphatic vascular invasion was detected in 20 patients (58.8%), neuronal invasion in 4 cases (11.8%), also 26 (76.5%) patients had five-years survival. Correlation between CD133 expression and clinicopathological factors features Based on our pathologic findings, expression of CD133 marker was positive in 25(73.5) patients and negative in 9 (26.5) patients. Associations between CD133 expression rate and clinic-pathological variables are presented in Table 2 . Table 2 Association of CD133 expression with clinic-pathological variables CD133 expression ( %) Variables No. 0 1+ 2+ 3+ p -value Gender Male 15 4 (55.6) 0 6(75.0) 4(33.3) 0.044 Female 19 8 (66.7) 2 (25.0) 5 (100) 4 (44.4) Age 34 55.77 ± 17.15 470 ± 14.47 67.25 ± 10.52 59.83 ± 16.114 0.267 Tumor Location Colon 16 4(44.4) 2(40.0) 6(75.0) 4(33.3) 0.182 Rectum 10 2 (22.3) 3 (60.0) 2 (25.0) 3 (25.0) Sekom 8 3(33.3) 0 0 5(41.7) Tumor Stage I 9 3 (33.3) 4 (80.0) 2 (25.0) 0 0.011 II 11 4 (44.4) 1 (20.0) 3 (37.5) 3 (25.0) III 14 9 (75.0) 3 (37.5) 0 2 (22.3) Tumor grade High 4 0 0 0 4 ( 33.3) 0.05 Low 30 8 (66.7) 8 (100.0) 5 (100.0) 9 (100.0) Lymphatic invasion Positive 20 2 (22.2) 2 (40.0) 4 (50.0) 12 (100.0) 0.002 Negative 14 7 (77.8) 3 (60.0) 4(50.00 0 Neural invasion Positive 4 0 0 1 (12.5) 3 (25.0) 0.273 Negative 30 9 (75.0) 7 (87.5) 5 (100.0) 9 (100.0) 5 Years survival Positive 26 9 (100.0) 2 (40.0) 7 (87.5) 8 (66.7) 0.054 Negative 8 4 (33.3) 1 (12.5) 3 (60.0) 0 According to the results (shown in table II), there was a significant difference between the expression of CD133 with gender, tumor stage and lymphatic vascular invasion in the studied patients (P = 0.044, P = 0.011, P = 0.002, respectively). The highest CD133 expression was observed at 8 females (66.7%). Also, all cases with high CD133 expression had the detected lymphatic invasion (p = 0.002). In addition, the highest CD133 expression was observed in CRC cases in advanced tumor stages III. However, no significant correlation was found between the CD133 expressions and the other clinical factors such as: age of CRC patients (p = 0.267), tumor location (P = 0.182), tumor differentiation grade (P = 0.05), neural tissue invasion (p = 0.002) and 5 years’ survival (P = 0.054). The 5-year survival in 26 (66.7%) patients with high CD13 expression showed no significantly differences with patients without 5-year survival and low CD133 expression (p value = 0.054). Discussion Colon cancer, as the leading cause of cancer- related death is a significant public health concern due to its increasing rate in Iran [ 41 – 44 ]. Many specific immunohistochemical and PCR markers have been identified for detecting colon primary and metastatic tumors. CD133 is one of the immunohistochemical markers which was first observed in stem cells [ 45 – 48 ]. This marker has been suggested to be effective in cell proliferation and differentiation of various cancers including colon [ 49 – 51 ]. Therefore, in this study, we purposed to evaluate the expressions of C D133 using immunohistochemically staining, also its relationship to clinic pathological features as well as the prognostic indicator in colon cancer. Our study demonstrated that the CD133 expression was significantly correlated with CRC histopathology such as gender tumor, stage and lymphatic vascular invasion (P = 0.044, P = 0.011, P = 0.002, respectively). The higher CD133 expression was found at advance tumor stage (III). But it did not affect other factors such as age, five-year survival and tumor location in this study. In the line with our study, there was no significant difference between positive and negative CD133 and tumor location in several previous studies [ 16 , 31 , 34 , 52 – 54 ]. In contrary, our study result was different from a study in which CD133 expression was higher in the rectum colon [ 45 ]. Also, similar to our result, in a study done by Kojima on samples from 189 patients with different stages of CRC using IHC, they concluded that CD133 overexpression occurred mainly in well- to moderately-differentiated tumors and was not correlated with recurrence-free survival [ 34 ]. However, there was conflicting results with our study regarding the expression of CD133 IHC staining in CRC and it relationship with the clinic pathological factors [68,55]. Many studies have demonstrated that CD133 expression is correlated with survival, recurrence, metastases and chemotherapy resistance, and most studies support the hypothesis that high D133 expression is a poor prognostic marker. [ 56 – 57 ]. In addition, in contrast to our results, Pitule et al. showed that patients with high CD133 expression had longer disease-free survival interval [ 40 ]. While our study as well as some other studies reported that CD133 was not significantly correlated with the survival time (5 years) [ 58 – 60 ]. Another study performed by Park on CRC 303 patients using immunhistchemical staining, showed that CD133 expression in CRC was significantly associated with five-year survival and tumor stage, but no with other clinical factors such as gender which was significantly related in our study. Moreover, they suggested Cd133 expression may consider as a more potential biomarker for prognosis or a high-risk feature in the stage II CRC patients [ 61 ]. This result was not consistent with our results. Contrary to our results, the results of a meta-analysis of 37 studies done by Huang in 2018 demonstrated that higher CD133 expression was positively correlated with shorter overall survival, lymphatic vascular invasion, distant metastasis and poorer prognosis in CRC patients. They also assumed that this low 5-year survival rate that might play an important role in the progression of colorectal cancer in CRC patients, was due to sex hormone, genetic and epigenetic factors that is affected by the environment and lifestyle and a variety of therapies [ 37 ]. In Rey study on 118 patients in 2020, CD133 expression was significantly associated with the tumor location (p = 0.002), but not with other clinic pathological factors such as gender, age, body mass. Also, the tumor location had impact on the survival of CRC patients. This was in line with two previous studies [ 45 , 49 , 51 ] but no with our study. In immunohistochemistry study of Kazama on 200 endoscopically resected colorectal polyps and 20 normal mucosae, they demonstrated that CD133 expression was associated with only the degree of tumor differentiation and tumor size but not with gender, age, tumor location. Apart from two factors including age and tumor location, the statistical result of the gender parameter was consistent with our results. So, CD133 might play an important role in tumor development. Finally, the difference of our results with other researches may be due to different geographical area or race. Conclusions In Conclusion, based on our findings there were a statistically significant difference between CD133 expression with gender, tumor stage III and lymph vascular invasion, but no other factors, although they showed some non-significantly differences. Therefore, the CD133 expression is potential to be used as a CRC prognostic stem cell marker for colon cancer. There was a limitation in our study. A larger number of cases is needed to achieve higher statistical power to detect significant differences. Declarations Ethics approval and consent to participate This study was approved by Ethical Committee of Kashan University of Medical Sciences, All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and national research committee and with the 1964 Helsinki Declaration and its later amendments. Consent for publication All study process was presented to patients and they were reassured about confidentiality of their records, they were requested to present their written consent of participation in the study. Availability of data and material The primary data for this study is available from the authors on direct request. Competing interests The authors declare no conflict of interest. Funding This work was supported by Kashan University of Medical Science, Kashan, Iran. We also thank the Deputy of Research and Technology, Ministry of Health and Medical Education of Iran for research grant support. Authors' contributions MT and HE were responsible for the study conception and design. MT and HE performed data collection. HE and HHK preparing the first draft of the manuscript. HHK made critical revisions to the paper for important intellectual content. Acknowledgements At the end of this study, authors of the current study represent their most appreciation to officials and nurses of Shahid Beheshti Hospital affiliated at Kashan University of Medical Sciences, officials of microbial laboratories in special. References Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 2010;127:2893–917. Jemal A, Center MM, DeSantis C, Ward EM. Global patterns of cancer incidence and mortality rates and trends. Cancer Epidemiol Biomarkers Prev 2010;19:1893–907 Siegel RL, Miller KD, Jemal A. Cancer statistics, 2018. CA Cancer J Clin. 2018; 68:7–30. https://doi.org/10.3322/ caac.21442. Kolahdoozan S, Sadjadi A, Radmard AR, Khademi H. Five common cancers in Iran. Arch Iran Med 2010; 13:143–46 Munro MJ, Wickremesekera SK, Peng L, Tan ST, Itinteang T. Cancer stem cells in colorectal cancer: a review. J of Clin Pathol 2018; 71:110–6 Dalerba P, Dylla SJ, Park IK, Liu R, Wang X, Cho RW, et al. Phenotypic characterization of human colorectal cancer stem cells. Proc Natl Acad Sci U S A 2007;104:10158–63 Ricci-Vitiani L, Lombardi DG, Pilozzi E, et al. Identification and expansion of human colon-cancer-initiating cells. Nature 2007;445: 111–5. Zhou Y, Xia L, Wang H, Oyang L, Su M, Liu Q, Lin J, Tan S, Tian Y, Liao Q, Cao D. Cancer stem cells in progression of colorectal cancer. Oncotarget 2018; 9:33403–15. Andre T, Boni C, Navarro M, Tabernero J, Hickish T, Topham C, et al. Improved overall survival with oxaliplatin, fluorouracil, and leucovorin as adjuvant treatment in stage II or III Dick JE. Looking ahead in cancer stem cell research. Nat Biotechnol 2009;27:44–6. Chen S, Huang EH. The colon cancer stem cell microenvironment holds keys to future cancer therapy. J Gastrointest Surg 2014; 18:1040–8. Gangemi R, Paleari L, Orengo AM, Cesario A, Chessa L, Ferrini S, et al. Cancer stem cells: a new paradigm for understanding tumor growth and progression and drug resistance. Curr Med Chem 2009; 16:1688–703. Corbeil D, Röper K, Hellwig A, Tavian M, Miraglia S, Watt SM, Simmons PJ, Peault B, Buck DW, Huttner WB. The human AC133 hematopoietic stem cell antigen is also expressed in epithelial cells and targeted to plasma membrane protrusions. J Biol Chem 2000; 275:5512–20 Shmelkov SV, St Clair R, Lyden D, Rafii S. AC133/CD133/Prominin-Int J Biochem Cell Biol. 2005;37:715e719.Ferrandina G, onanno G, Pierelli L, et al. Expression of CD133-1 and CD133-2 in ovarian cancer. Int J Gynecol Cancer. 2008;18:506e514 Yin AH, Miraglia S, Zanjani ED, et al. AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood 1997;90:5002–12 Yin AH, Miraglia S, Zanjani ED, Almeida-Porada G, Ogawa M, Leary AG, Olweus J, Kearney J, Buck DW: AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood 1997, 90:5002–5012. Uchida N, Buck DW, He D, Reitsma MJ, Masek M, Phan TV, Tsukamoto AS, Gage FH, Weissman IL: Direct isolation of human central nervous system stem cells. Proc Natl Acad Sci U S A 2000, 97:14720–14725. Collins AT, Berry PA, Hyde C, Stower MJ, Maitland NJ: Prospective identification of tumorigenic prostate cancer stem cells. Cancer Res 2005, 65:10946–10951. Ricci-Vitiani L, Lombardi DG, Pilozzi E, Biffoni M, Todaro M, Peschle C,De Maria R: Identification and expansion of human colon- cancer-initiating cells. Nature 2007, 445:111–115\ Suetsugu A, Nagaki M, Aoki H, Motohashi T, Kunisada T,Moriwaki H. Characterization of CD133 hepatocellular carcinoma cells as cancer stem/progenitor cells. Biochem Biophys Res Communes. 2006;351:820e824. O’Brien CA, Pollett A, Gallinger S, Dick JE. A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature. 2007;445:106e110. Ricci-Vitiani L, Lombardi DG, Pilozzi E, et al. Identification and expansion of human colon-cancer-initiating cells. Nature. 2007;445:111e115 L. Ricci-Vitiani, D. G. Lombardi, E. Pilozzi et al., “Identification and expansion of human colon-cancer-initiating cells,” Nature, vol. 445, no. 7123, pp. 111–115, 2007. C. A. O’Brien, A. Pollett, S. Gallinger, and J. E. Dick, “A human colon cancer cell capable of initiating tumour growth in immunodeficient mice,” Nature, vol. 445, no. 7123, pp. 106–110, 2007. Eramo A, Lotti F, Sette G, Pilozzi E, Biffoni M, Di Virgilio A, et al. Identification and expansion of the tumorigenic lung cancer stem cell population.Cell Death & Differentiation, 2008. 15(3): 504–514. icci-Vitiani L, Lombardi DG, Pilozzi E, Biffoni M, Todaro M, Peschle C, et al.Identification and expansion of human coloncancer-initiating cells. Nature, 2007. 445 (7123):111–115. Singh SK, Hawkins C, Clarke ID, Squire JA, Bayani J, Hide T, et al. Identification of human brain tumour initiating cells. Nature, 2004. 432 (7015): 396–401.\ Cheng J-X, Liu B-L, Zhang X. How powerful is CD133 as a cancer stem cell marker in brain tumors? Cancer Treat Rev. 2009;35(5):403–8. [DOI: 10.1016/j.ctrv.2009.03.002] [PMID] Yin AH, Miraglia S, Zanjani ED, et al. AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood1997;90:5002^12. Bonanno G, Perillo A, Rutella S, et al. Clinical isolation and functional characterization of cord blood CD133 + hematopoietic progenitor cells. Transfusion 2004;44:1087 ^ 97 Bonanno G,Mariotti A, Procoli A, et al. Human cord blood CD133 + cells immunoselected by a clinicalgrade apparatus differentiate in vitro into endothelialand cardiomyocyte-like cells. Transfusion 2007;47: 280^9. Park SJ, Ahn TS, Cho SW, Kim CJ, Jung DJ, Son MW, et al. Stromal-cell-derived Factor 1-alpha Promotes Tumor Progression in Colorectal Cancer. J Korean Soc Coloproctol 2012; 28: 27–34. LaBarge MA, Bissell MJ. Is CD133 a marker of metastatic colon cancer stem cells? J Clin Invest 2008; 118: 2021–24. Kojima M, Ishii G, Atsumi N, Fujii S, Saito N, Ochiai A. Immunohistochemical detection of CD133 expression in colorectal cancer: a clinicopathological study. Cancer Sci. 2008;99: 1578e1583. Horst D, Kriegl L, Engel J, Kirchner T, Jung A. CD133 expression is an independent prognostic marker for low survival in colorectal cancer. Br J Cancer. 2008;99:1285e1289. Wang K, Xu J, Zhang J, Huang J. Prognostic role of CD133 expression in colorectal cancer: a meta-analysis. BMC Cancer. 2012 Dec 5;12:573. Huang R, Mo D Wu J, Ai H, Lu Y.CD133 expression correlates with clinicopathologic features and poor prognosis of colorectal cancer patients: An updated meta-analysis of 37 studies. Medicine (Baltimore). 2018 Jun;97(23):e10446. Kim ST, Sohn I, DO IG, et al. Transcriptome analysis of CD133-positive stem cells and prognostic value of survivin in colorectal cancer. Cancer Genomics Proteomics. 2014 Sep-Oct;11(5):259–66. Reggiani Bonetti L, Migaldi M, Caredda E, et al. Increased expression of CD133 is a strong predictor of poor outcome in stage I colorectal cancer patients. Scand J Gastroenterol. 2012 Oct;47(10):1211–7. Pitule P, Cedikova M, Daum O, et al. Immunohistochemical detection of cancer stem cell related markers CD44 and CD133 in metastatic colorectal cancer patients. Biomed Res Int. 2014;2014:432139. Hosseini S, Izadpanah A, Yarmohammadi H. Epidemiological changes in colorectal cancer in Shiraz, Iran: 1980–2000. ANZ J Surg 2004; 74:547–49. Ansari R, Mahdavinia M, Sadjadi A, Nouraie M, Kamangar F, Bishehsari F, et al. Incidence and age distribution of colorectal cancer in Iran: results of a population-based cancer registry. Cancer Lett 18 2006;240:143–47. Foroutan M, Rahimi N, Tabatabaeifar M, Darvishi M, Hashemi M, Hosseinpanah F, et al. Clinical features of colorectal cancer in Iran: A 15-year review. Dig Dis 2008; 9:225–27. Pahlavan PS, Jensen K. A short impact of epidemiological features of colorectal cancer in Iran. Tumori 2005; 91:291–94. Hong I, Hong SW, Chang YG, Lee WY, Lee B, Kang YK, Kim YS, Paik IW, Lee H. Expression of the cancer stem cell markers CD44 and CD133 in colorectal cancer: an immunohistochemical staining analysis.Ann Coloproctol 2015; 31:84–91. Hongo K, Kazama S, Sunami E, Tsuno NH, Takahashi K, Nagawa H, Kitayama J. Immunohistochemical detection of CD133 is associated with tumor regression grade after chemoradiotherapy in rectal cancer. Med Oncol 2012; 29:2849–57. Kojima M, Ishii G, Atsumi N, Fujii S, Saito N, Ochiai A. Immunohistochemical detection of CD133 expression in colorectal cancer: a clinicopathological study. Cancer Sci 2008; 99:1578–83. Pitule P, Cedikova M, Daum O, Vojtisek J, Vycital O, Hosek P, et al. Immunohistochemical detection of cancer stem cell related markers CD44 and CD133 in metastatic colorectal cancer patients. Biomed Res Int 2014;2014:432139. Chen S, Song X, Chen Z, et al. CD133 expression and the prognosis of colorectal cancer: a systematic review and metaanalysis. PLoS One. 2013;8:e56380. Schneider M, Huber J, Hadaschik B, Siegers GM, Fiebig H-H, Schüler J. Characterization of colon cancer cells: a functional approach characterizing CD133 as a potential stem cell marker. BMC cancer. 2012;12(1):1–11. Yang Z-L, Zheng Q, Yan J, Pan Y, Wang Z-G. Upregulated CD133 expression in tumorigenesis of colon cancer cells. World journal of gastroenterology: WJG. 2011;17(7):932. Ricci-Vitiani L, LombardiDG, Pilozzi E, et al. Identification and expansion of human colon-cancer-initiating cells.Nature 2007;445:111^5. \ Horst D, Kriegl L, Engel J, Kirchner T, Jung A. Prognostic significance of the cancer stem cell markers CD133, CD44, and CD166 in colorectal cancer. Cancer Invest 2009; 27:844–50. Huang R, Mo D, Wu J, Ai H, Lu Y. CD133 expression correlates with clinicopathologic features and poor prognosis of colorectal cancer patients: An updated meta-analysis of 37 studies. Medicine. 2018;97(23). Nosrati A, Naghshvar F, Maleki I, Salehi F. Cancer stem cells CD133 and CD24 in colorectal cancers n Northern Iran. Gastroenterol Hepatol Bed Bench 2016; 9:132–9 Iinuma H, Watanabe T, Mimori K, et al. Clinical significance of circulating tumor cells, including cancer stem-like cells, in peripheral blood for recurrence and prognosis in patients with Dukes’ stage B and C colorectal cancer. J Clin Oncol 2011;29:1547–55. Jao SW, Chen SF, Lin YS, et al. Cytoplasmic CD133 expression is a reliable prognostic indicator of tumor regression after neoadjuvant concurrent chemoradiotherapy in patients with rectal cancer. Ann Surg Oncol 2012;19:3432–40. Choi D, Lee HW, Hur KY, Kim JJ, Park GS, Jang SH, et al. Cancer stem cell markers CD133 and CD24 correlate with invasiveness and differentiation in colorectal adenocarcinoma. World J Gastroenterol 2009;15:2258–64. Lugli A, Iezzi G, Hostettler I, Muraro MG, Mele V, Tornillo L, et al. Prognostic impact of the expression of putative cancer stem cell markers CD133, CD166, CD44s, EpCAM, and ALDH1 in colorectal cancer. Br J Cancer 2010;103:382–90. Zhou F, Mu YD, Liang J, Liu ZX, Chen HS, Zhang JF. Expression and prognostic value of tumor stem cell markers ALDH1 and CD133 in colorectal carcinoma. Oncol Lett 2014;7:507–12. Park YY, An CH, Oh ST, Chang ED, Lee J. Expression of CD133 is associated with poor prognosis in stage II colorectal carcinoma. Medicine. 2019;98(32). 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-1538147","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":98402177,"identity":"e7b25f19-014d-4552-b4ff-6b0e15049ea5","order_by":0,"name":"Mohammad Tolooei","email":"","orcid":"","institution":"Kashan University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Mohammad","middleName":"","lastName":"Tolooei","suffix":""},{"id":98402178,"identity":"4e9eb3b1-dfb1-4c63-b16c-63595be8df44","order_by":1,"name":"Hassan Ehteram","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA9klEQVRIiWNgGAWjYDACCRS6AoiZmRsIaGFG1nIGpIWRFC2MbWASvxZ+6f6DjytqtjFItp8x3fhzXm00fztQy4+KbTi1SM45zGx45thtBmmeHLPbvNuO5844zNjA2HPmNk4tBjeS2SQb2G4zyDEAtTBuO5bbANTCzNiGW4v9jWT2nw3/gFr435jd/DnnWO58QloMJJLZGBuBCqQlcsxu8DbU5G4gpEXiRrKxZGPfbR7JGc/KbvMcO5C7EajlID6/8M9IfPix4dttOYnzydtu/qipy513/vDBBz8qcGuBAR4ofRhMHiCoHgnUkaJ4FIyCUTAKRggAAGXZW3nGvvzYAAAAAElFTkSuQmCC","orcid":"","institution":"Kashan University of Medical Sciences","correspondingAuthor":true,"prefix":"","firstName":"Hassan","middleName":"","lastName":"Ehteram","suffix":""},{"id":98402179,"identity":"290f783d-207b-4cec-9247-d6a8f1dc94df","order_by":2,"name":"Hamed Haddad Kashani","email":"","orcid":"","institution":"Kashan University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Hamed","middleName":"Haddad","lastName":"Kashani","suffix":""}],"badges":[],"createdAt":"2022-04-08 16:14:15","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-1538147/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-1538147/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":20454183,"identity":"859bf768-088e-47c8-90f1-d86fb6273ceb","added_by":"auto","created_at":"2022-04-18 14:22:18","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":366247,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1538147/v1/640d7f4c-1a27-4856-9dd1-1778a3124267.pdf"},{"id":20454182,"identity":"24788d76-6044-4143-af48-47b6bfdf39a2","added_by":"auto","created_at":"2022-04-18 14:22:17","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":366247,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1538147/v1/530e0eb2-c22b-45dd-a3fd-040ef5ea5a7e.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Expression and Prognostic Significance of Stem Cell Marker CD133 in Survival Rate of Colon Cancer Patients","fulltext":[{"header":"Introduction","content":"\u003cp\u003eColon cancer is one of the most prevalent cancers in the world and a major cause of cancer-related mortality [\u003cspan additionalcitationids=\"CR2\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. In Iran, colon cancer is the third most common cancer in men and fourth common cancer in women [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. It is suggested that the highly tumorigenic, chemotherapy resistant affect survival rate in colon cancer [\u003cspan additionalcitationids=\"CR6 CR7\" citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Targeted therapies have promising potential improvement of survival in colon cancer patients [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Recently, cancer stem cells (CSCs) have been known due to their ability of self-renew, self-repair and their role in the tumor initiation, metastasis, therapeutic resistance and recurrence [\u003cspan additionalcitationids=\"CR11\" citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Different markers have been also found to be expressed on the surface of CSCs among which CD133 has been paid the special attention. The CD133/prominin-1 is a 120-kDa 5-transmembrane domain antigen on chromosome 4p15.32 and mainly localized in membrane protrusions [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. CD133 first discovered in hematopoietic stem and progenitor cells by Yin et al [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Previous studies identified CD133 as a putative CSC marker in cancer cells [\u003cspan additionalcitationids=\"CR17 CR18\" citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e] with ability to initiate tumor growth [\u003cspan additionalcitationids=\"CR21 CR22 CR23\" citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. This CSC marker now widely used to identify cancer stem cells in many cancers such as brain, colon, and lung cancers [\u003cspan additionalcitationids=\"CR26 CR27\" citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. CD133 is normally expressed by primitive hematopoietic stem cells from adult blood and bone narrow, human fetal liver and stem cells in cord blood and peripheral blood, endothelial progenitor cells, epithelial cells and neural and glial stem cells, [\u003cspan additionalcitationids=\"CR30\" citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Several studies have suggested that high CD133 expression is associated with the clinic-pathologic characteristics such as poor prognosis, distant metastasis, survival rate and even chemotherapy resistance in colon cancer patients So, as a CRC-initiating CSC marker, CD133 expression can be used for developing a novel therapeutic approaches in colon cancer [\u003cspan additionalcitationids=\"CR33 CR34 CR35 CR36 CR37 CR38 CR39\" citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. In this study, we investigated the association between the expression and prognostic significance of the CSC marker CD133 with clinic-pathologic features of CRC patients including survival rate, using the immunohistochemistry (IHC) method.\u003c/p\u003e"},{"header":"Methods And Materials","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eTissue samples\u003c/h2\u003e \u003cp\u003eIn this study, 34 formalin- fixed-paraffinembedded (FFPE) tumor tissues were selected from 34 patients underwent surgical resection from 2008\u0026ndash;2019 in Shahid Beheshti Hospital in Kashan, Iran. Written consent was obtained from all subjects and the study was approved by the Research Ethical Committee of Kashan University of Medical Sciences, Iran. All clinic-pathologic data were verified using the hematoxylin and eosin (H\u0026amp;E) stained pathologic slides, patients\u0026rsquo; medical files and pathology record. Tumors were classified according to the American Joint Committee on Cancer (AJCC) and the International Union Against Cancer (UICC) TNM criteria [93]. Clinic-pathologic parameters were included age, gender, tumor location, tumor stage, tumor differentiation grade, lymphatic and neural tissue invasion, 5 years\u0026rsquo; survival and tumor recurrence. A colon cancer with detected positive immunostaining for CD133 considered as a positive control. Cases and controls were examined by two expert pathologists. Positive reaction was defined by the presence of at least 10% of cancer cells displaying either membranous and/or cytoplasmic staining. A number of tumor samples were excluded from the study due to inappropriate paraffin blocks, low quality staining and unavailable data. Also, the small size specimens prepared by biopsy which was inadequate for performing histopathological examinations were excluded.\u003c/p\u003e \u003cdiv id=\"Sec4\" class=\"Section3\"\u003e \u003ch2\u003eCD133 Immuno-histochemical staining\u003c/h2\u003e \u003cp\u003eThe expression level of CD133 was determined in CRC sections by the Immunohistochemical staining (IHC) method as follows: Formalin-fixed, paraffin-embedded CRC tissue blocks were sectioned at 5-\u0026micro;m thickness for IHC staining. All slides were deparaffinized with xylene, and rehydrated in a graded ethylic alcohol (50%, 70%, 80% and 100%) followed by distilled water. After washing 3 times with phosphate-buffered saline (PBS) buffer every 5 minutes, endogenous peroxidase was inactive by immersing the sections in 1-0.1% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e for 5min. Then, for heat-induced antigen unmasking, sample tissues were treated in sodium citrate buffer (pH:6.0) at 95\u0026deg;C for 5 minutes in a microwave oven and allowed to cool for 1 hour at room temperature. CD133 expression was identified and visualized after reaction with primary Anti CD133 antibody (1:100) at 4˚C for one night. After washing the slides three times with PBS, a secondary antibody (peroxides- conjugated) was used to detect the primary antibody, and slides were incubated for 30 minutes in room temperature. Again, after three times washing in PBS for every 5 minutes, samples were incubated with 3, 3\u0026acute;-diaminobenzidine tetrahydrochloride-chromogen (DAB) for 1\u0026ndash;5 minutes for visualizing immunostaining and finally counterstained with hematoxylin dye and dehydrated in ethylic alcohol serial dilution (60%,80% and 100%) and cleared in xylene (100%). Our negative control was the tissue that was covered with just buffer without the primary antibody. Prepared specimens were studied using light microscope. Imaging analysis of immunohistochemical staining of the specimens was evaluated by one pathologist without knowledge about the patients' clinic-pathological characteristics. The CD133 expression was assessed by observing the appearance of yellow to brown color in the cytoplasmic and membrane of glandular epithelial cells. A score of less than 10% CD133 cells (low CD133 expression) considered as negative and more than 10% CD133 cells (high CD133 expression) as positive, respectively. The staining intensity was evaluated according a ranging from non-stained (negative) to strong (positive): 0; non-staining, +\u0026thinsp;1; weakly staining, +\u0026thinsp;2; moderate staining, +\u0026thinsp;3; strong staining.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eAll statistical analyses were performed using SPSS software version 20 (SPSS Inc., Chicago, IL., USA). The results were expressed as the means\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation. The statistical significance of differences between immunohistochemical CD133 expression levels and clinicopathological features of CRC patients were evaluated using Chi-square and one-way ANOVA, or Mann-Whitney test or Kruskal-Wallis parametric bread tests (based on the results of Kolmogorov-Smirnov test), as appropriate. Survival was evaluated using the Kaplan-Meier method with a log-rank test. \u003cem\u003ep\u003c/em\u003e-values of \u0026lt;\u0026thinsp;0.05 were considered statistically significant.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eThis study included 34 tissue samples from 15 (44.1) male patients and 19 (55.9%) female patients with a median age of 62 (\u0026plusmn;\u0026thinsp;15.35) years at the time of diagnosis. The distribution of CRC patients with colon cancer according to demographic features is presented in Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eCharacteristics of 38 CRC patients and their prognostic factors.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eVariable ( No.%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGender\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15 (44.1)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e19 ( 55.9)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAge (mean) (\u0026plusmn;\u0026thinsp;SD) (years)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e62 (\u0026plusmn;\u0026thinsp;15.35)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTumor location\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAscendance colon\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14 (429.4%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTransverse\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1(3.0%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSekom\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6 (18.2%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSigmoid 8 (2.2%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8(24.2%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDescending\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1 (3.0%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRectum colon\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2 (6.1%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDariche ileocecal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2(6.1%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eHistopathology Grade\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWell-differentiated ( G1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e21 (61.8%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eModerately differentiated (G2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e11 (32.4%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePoorly differentiated (G3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2(5.8%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCD133 expression\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHigh Level\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e12 ( 35.3%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLow Level\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9 (26.5%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e5 years\u0026rsquo; survival\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePositive\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e26 (76.5%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNegative\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8 ( 23.5%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLymphatic invasion\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePositive\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20 ( 58.8%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNegative\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14 (41.2%0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTumor stage\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9 (26.5%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e7 ( 20.6)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4 (11.8%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1 (2.9%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8 (23.5%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5 (14.7%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNeuronal invasion\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePositive\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4 (11.8%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNegative\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e30 (88.2%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe pathological results showed (as shown at Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) that the most tumors were located in ascendance colon 13 (39.4%) and in Sigmoid 8 (2.2%), while lowest tumor location was descending 1 (3.0%) and transverse 1 (3.0%). Also based on tumor differentiation degree, there were 21 (61.8%) well-differentiated (G1), 11 (32.4%) moderated differentiated (G2) and 2(5.8%) poor differentiation (G3) tumors. The highest prevalence of tumor stages was stage I (26.5%) and stage 3b (23.5%), while the lowest frequency was observed at stage 3a (2.9%). Moreover, lymphatic vascular invasion was detected in 20 patients (58.8%), neuronal invasion in 4 cases (11.8%), also 26 (76.5%) patients had five-years survival.\u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eCorrelation between CD133 expression and clinicopathological factors features\u003c/h2\u003e \u003cp\u003eBased on our pathologic findings, expression of CD133 marker was positive in 25(73.5) patients and negative in 9 (26.5) patients. Associations between CD133 expression rate and clinic-pathological variables are presented in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAssociation of CD133 expression with clinic-pathological variables\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"7\" nameend=\"c7\" namest=\"c1\"\u003e \u003cp\u003eCD133 expression ( %)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eVariables\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eNo.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e0\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e1+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e2+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e3+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003ep\u003c/span\u003e\u003cb\u003e-value\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"6\" nameend=\"c6\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGender\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4 (55.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6(75.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4(33.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003e0.044\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8 (66.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2 (25.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5 (100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4 (44.4)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAge\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e55.77\u0026thinsp;\u0026plusmn;\u0026thinsp;17.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e470\u0026thinsp;\u0026plusmn;\u0026thinsp;14.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e67.25\u0026thinsp;\u0026plusmn;\u0026thinsp;10.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e59.83\u0026thinsp;\u0026plusmn;\u0026thinsp;16.114\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.267\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTumor Location\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"6\" nameend=\"c7\" namest=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eColon\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4(44.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2(40.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6(75.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4(33.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003e\u003cb\u003e0.182\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRectum\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2 (22.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3 (60.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2 (25.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3 (25.0)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSekom\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3(33.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5(41.7)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTumor Stage\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"6\" nameend=\"c7\" namest=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3 (33.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4 (80.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2 (25.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003e\u003cb\u003e0.011\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eII\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4 (44.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1 (20.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3 (37.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3 (25.0)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIII\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e9 (75.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3 (37.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2 (22.3)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTumor grade\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"6\" nameend=\"c7\" namest=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHigh\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4 ( 33.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003e0.05\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLow\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8 (66.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8 (100.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5 (100.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9 (100.0)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLymphatic invasion\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c7\" namest=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePositive\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2 (22.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2 (40.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4 (50.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e12 (100.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003e0.002\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNegative\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7 (77.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3 (60.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4(50.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNeural invasion\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c7\" namest=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePositive\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1 (12.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3 (25.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003e0.273\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNegative\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e9 (75.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7 (87.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5 (100.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9 (100.0)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"7\" nameend=\"c7\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003e5 Years survival\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePositive\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e9 (100.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2 (40.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e7 (87.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8 (66.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003e0.054\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNegative\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4 (33.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1 (12.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3 (60.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eAccording to the results (shown in table II), there was a significant difference between the expression of CD133 with gender, tumor stage and lymphatic vascular invasion in the studied patients (P\u0026thinsp;=\u0026thinsp;0.044, P\u0026thinsp;=\u0026thinsp;0.011, P\u0026thinsp;=\u0026thinsp;0.002, respectively). The highest CD133 expression was observed at 8 females (66.7%). Also, all cases with high CD133 expression had the detected lymphatic invasion (p\u0026thinsp;=\u0026thinsp;0.002). In addition, the highest CD133 expression was observed in CRC cases in advanced tumor stages III. However, no significant correlation was found between the CD133 expressions and the other clinical factors such as: age of CRC patients (p\u0026thinsp;=\u0026thinsp;0.267), tumor location (P\u0026thinsp;=\u0026thinsp;0.182), tumor differentiation grade (P\u0026thinsp;=\u0026thinsp;0.05), neural tissue invasion (p\u0026thinsp;=\u0026thinsp;0.002) and 5 years\u0026rsquo; survival (P\u0026thinsp;=\u0026thinsp;0.054). The 5-year survival in 26 (66.7%) patients with high CD13 expression showed no significantly differences with patients without 5-year survival and low CD133 expression (p value\u0026thinsp;=\u0026thinsp;0.054).\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eColon cancer, as the leading cause of cancer- related death is a significant public health concern due to its increasing rate in Iran [\u003cspan additionalcitationids=\"CR42 CR43\" citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. Many specific immunohistochemical and PCR markers have been identified for detecting colon primary and metastatic tumors. CD133 is one of the immunohistochemical markers which was first observed in stem cells [\u003cspan additionalcitationids=\"CR46 CR47\" citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. This marker has been suggested to be effective in cell proliferation and differentiation of various cancers including colon [\u003cspan additionalcitationids=\"CR50\" citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. Therefore, in this study, we purposed to evaluate the expressions of C D133 using immunohistochemically staining, also its relationship to clinic pathological features as well as the prognostic indicator in colon cancer. Our study demonstrated that the CD133 expression was significantly correlated with CRC histopathology such as gender tumor, stage and lymphatic vascular invasion (P\u0026thinsp;=\u0026thinsp;0.044, P\u0026thinsp;=\u0026thinsp;0.011, P\u0026thinsp;=\u0026thinsp;0.002, respectively). The higher CD133 expression was found at advance tumor stage (III). But it did not affect other factors such as age, five-year survival and tumor location in this study. In the line with our study, there was no significant difference between positive and negative CD133 and tumor location in several previous studies [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e, \u003cspan additionalcitationids=\"CR53\" citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. In contrary, our study result was different from a study in which CD133 expression was higher in the rectum colon [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. Also, similar to our result, in a study done by Kojima on samples from 189 patients with different stages of CRC using IHC, they concluded that CD133 overexpression occurred mainly in well- to moderately-differentiated tumors and was not correlated with recurrence-free survival [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. However, there was conflicting results with our study regarding the expression of CD133 IHC staining in CRC and it relationship with the clinic pathological factors [68,55]. Many studies have demonstrated that CD133 expression is correlated with survival, recurrence, metastases and chemotherapy resistance, and most studies support the hypothesis that high D133 expression is a poor prognostic marker. [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e]. In addition, in contrast to our results, Pitule et al. showed that patients with high CD133 expression had longer disease-free survival interval [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. While our study as well as some other studies reported that CD133 was not significantly correlated with the survival time (5 years) [\u003cspan additionalcitationids=\"CR59\" citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e]. Another study performed by Park on CRC 303 patients using immunhistchemical staining, showed that CD133 expression in CRC was significantly associated with five-year survival and tumor stage, but no with other clinical factors such as gender which was significantly related in our study. Moreover, they suggested Cd133 expression may consider as a more potential biomarker for prognosis or a high-risk feature in the stage II CRC patients [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e]. This result was not consistent with our results. Contrary to our results, the results of a meta-analysis of 37 studies done by Huang in 2018 demonstrated that higher CD133 expression was positively correlated with shorter overall survival, lymphatic vascular invasion, distant metastasis and poorer prognosis in CRC patients. They also assumed that this low 5-year survival rate that might play an important role in the progression of colorectal cancer in CRC patients, was due to sex hormone, genetic and epigenetic factors that is affected by the environment and lifestyle and a variety of therapies [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. In Rey study on 118 patients in 2020, CD133 expression was significantly associated with the tumor location (p\u0026thinsp;=\u0026thinsp;0.002), but not with other clinic pathological factors such as gender, age, body mass. Also, the tumor location had impact on the survival of CRC patients. This was in line with two previous studies [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e, \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e] but no with our study. In immunohistochemistry study of Kazama on 200 endoscopically resected colorectal polyps and 20 normal mucosae, they demonstrated that CD133 expression was associated with only the degree of tumor differentiation and tumor size but not with gender, age, tumor location. Apart from two factors including age and tumor location, the statistical result of the gender parameter was consistent with our results. So, CD133 might play an important role in tumor development. Finally, the difference of our results with other researches may be due to different geographical area or race.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eIn Conclusion, based on our findings there were a statistically significant difference between CD133 expression with gender, tumor stage III and lymph vascular invasion, but no other factors, although they showed some non-significantly differences. Therefore, the CD133 expression is potential to be used as a CRC prognostic stem cell marker for colon cancer. There was a limitation in our study. A larger number of cases is needed to achieve higher statistical power to detect significant differences.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003e\u003cem\u003eEthics approval and consent to participate\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was approved by\u0026nbsp;Ethical Committee of Kashan University of Medical Sciences,\u0026nbsp;All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and national research committee and with the 1964 Helsinki Declaration and its later amendments.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eConsent for publication\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll study process was presented to patients and they were reassured about confidentiality of their records, they were requested to present their written consent of participation in the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eAvailability of data and material\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe primary data for this study is available from the authors on direct request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eCompeting interests\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eFunding\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by Kashan University of Medical Science, Kashan, Iran. We also thank the\u0026nbsp;Deputy of Research and Technology, Ministry of Health and Medical Education\u0026nbsp;of Iran for research grant support.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eAuthors\u0026apos; contributions\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMT and HE were responsible for the study conception and design. MT and HE\u0026nbsp;performed data collection. HE and HHK preparing the first draft of the manuscript. HHK made critical revisions to the paper for important intellectual content.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eAcknowledgements\u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAt the end of this study, authors of the current study represent their most appreciation to officials and nurses of Shahid Beheshti Hospital affiliated at Kashan University of Medical Sciences, officials of microbial laboratories in special.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eFerlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 2010;127:2893\u0026ndash;917.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJemal A, Center MM, DeSantis C, Ward EM. Global patterns of cancer incidence and mortality rates and trends. Cancer Epidemiol Biomarkers Prev 2010;19:1893\u0026ndash;907\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSiegel RL, Miller KD, Jemal A. Cancer statistics, 2018. CA Cancer J Clin. 2018; 68:7\u0026ndash;30. https://doi.org/10.3322/ caac.21442.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKolahdoozan S, Sadjadi A, Radmard AR, Khademi H. Five common cancers in Iran. Arch Iran Med 2010; 13:143\u0026ndash;46\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMunro MJ, Wickremesekera SK, Peng L, Tan ST, Itinteang T. Cancer stem cells in colorectal cancer: a review. J of Clin Pathol 2018; 71:110\u0026ndash;6\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDalerba P, Dylla SJ, Park IK, Liu R, Wang X, Cho RW, et al. Phenotypic characterization of human colorectal cancer stem cells. Proc Natl Acad Sci U S A 2007;104:10158\u0026ndash;63\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRicci-Vitiani L, Lombardi DG, Pilozzi E, et al. Identification and expansion of human colon-cancer-initiating cells. Nature 2007;445: 111\u0026ndash;5.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhou Y, Xia L, Wang H, Oyang L, Su M, Liu Q, Lin J, Tan S, Tian Y, Liao Q, Cao D. Cancer stem cells in progression of colorectal cancer. Oncotarget 2018; 9:33403\u0026ndash;15.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAndre T, Boni C, Navarro M, Tabernero J, Hickish T, Topham C, et al. Improved overall survival with oxaliplatin, fluorouracil, and leucovorin as adjuvant treatment in stage II or III\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDick JE. Looking ahead in cancer stem cell research. Nat Biotechnol 2009;27:44\u0026ndash;6.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen S, Huang EH. The colon cancer stem cell microenvironment holds keys to future cancer therapy. J Gastrointest Surg 2014; 18:1040\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGangemi R, Paleari L, Orengo AM, Cesario A, Chessa L, Ferrini S, et al. Cancer stem cells: a new paradigm for understanding tumor growth and progression and drug resistance. Curr Med Chem 2009; 16:1688\u0026ndash;703.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCorbeil D, R\u0026ouml;per K, Hellwig A, Tavian M, Miraglia S, Watt SM, Simmons PJ, Peault B, Buck DW, Huttner WB. The human AC133 hematopoietic stem cell antigen is also expressed in epithelial cells and targeted to plasma membrane protrusions. J Biol Chem 2000; 275:5512\u0026ndash;20\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShmelkov SV, St Clair R, Lyden D, Rafii S. AC133/CD133/Prominin-Int J Biochem Cell Biol. 2005;37:715e719.Ferrandina G, onanno G, Pierelli L, et al. Expression of CD133-1 and CD133-2 in ovarian cancer. Int J Gynecol Cancer. 2008;18:506e514\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYin AH, Miraglia S, Zanjani ED, et al. AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood 1997;90:5002\u0026ndash;12\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYin AH, Miraglia S, Zanjani ED, Almeida-Porada G, Ogawa M, Leary AG, Olweus J, Kearney J, Buck DW: AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood 1997, 90:5002\u0026ndash;5012.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eUchida N, Buck DW, He D, Reitsma MJ, Masek M, Phan TV, Tsukamoto AS, Gage FH, Weissman IL: Direct isolation of human central nervous system stem cells. Proc Natl Acad Sci U S A 2000, 97:14720\u0026ndash;14725.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCollins AT, Berry PA, Hyde C, Stower MJ, Maitland NJ: Prospective identification of tumorigenic prostate cancer stem cells. Cancer Res 2005, 65:10946\u0026ndash;10951.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRicci-Vitiani L, Lombardi DG, Pilozzi E, Biffoni M, Todaro M, Peschle C,De Maria R: Identification and expansion of human colon- cancer-initiating cells. Nature 2007, 445:111\u0026ndash;115\\\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSuetsugu A, Nagaki M, Aoki H, Motohashi T, Kunisada T,Moriwaki H. Characterization of CD133 hepatocellular carcinoma cells as cancer stem/progenitor cells. Biochem Biophys Res Communes. 2006;351:820e824.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eO\u0026rsquo;Brien CA, Pollett A, Gallinger S, Dick JE. A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature. 2007;445:106e110.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRicci-Vitiani L, Lombardi DG, Pilozzi E, et al. Identification and expansion of human colon-cancer-initiating cells. Nature. 2007;445:111e115\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eL. Ricci-Vitiani, D. G. Lombardi, E. Pilozzi et al., \u0026ldquo;Identification and expansion of human colon-cancer-initiating cells,\u0026rdquo; Nature, vol.\u0026nbsp;445, no. 7123, pp.\u0026nbsp;111\u0026ndash;115, 2007.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eC. A. O\u0026rsquo;Brien, A. Pollett, S. Gallinger, and J. E. Dick, \u0026ldquo;A human colon cancer cell capable of initiating tumour growth in immunodeficient mice,\u0026rdquo; Nature, vol.\u0026nbsp;445, no. 7123, pp.\u0026nbsp;106\u0026ndash;110, 2007.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEramo A, Lotti F, Sette G, Pilozzi E, Biffoni M, Di Virgilio A, et al. Identification and expansion of the tumorigenic lung cancer stem cell population.Cell Death \u0026amp; Differentiation, 2008. 15(3): 504\u0026ndash;514.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eicci-Vitiani L, Lombardi DG, Pilozzi E, Biffoni M, Todaro M, Peschle C, et al.Identification and expansion of human coloncancer-initiating cells. Nature, 2007. 445 (7123):111\u0026ndash;115.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh SK, Hawkins C, Clarke ID, Squire JA, Bayani J, Hide T, et al. Identification of human brain tumour initiating cells. Nature, 2004. 432 (7015): 396\u0026ndash;401.\\\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCheng J-X, Liu B-L, Zhang X. How powerful is CD133 as a cancer stem cell marker in brain tumors? Cancer Treat Rev. 2009;35(5):403\u0026ndash;8. [DOI:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ctrv.2009.03.002]\u003c/span\u003e\u003cspan address=\"10.1016/j.ctrv.2009.03.002]\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e [PMID]\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYin AH, Miraglia S, Zanjani ED, et al. AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood1997;90:5002^12.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBonanno G, Perillo A, Rutella S, et al. Clinical isolation and functional characterization of cord blood CD133 + hematopoietic progenitor cells. Transfusion 2004;44:1087 ^ 97\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBonanno G,Mariotti A, Procoli A, et al. Human cord blood CD133 + cells immunoselected by a clinicalgrade apparatus differentiate in vitro into endothelialand cardiomyocyte-like cells. Transfusion 2007;47: 280^9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePark SJ, Ahn TS, Cho SW, Kim CJ, Jung DJ, Son MW, et al. Stromal-cell-derived Factor 1-alpha Promotes Tumor Progression in Colorectal Cancer. J Korean Soc Coloproctol 2012; 28: 27\u0026ndash;34.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLaBarge MA, Bissell MJ. Is CD133 a marker of metastatic colon cancer stem cells? J Clin Invest 2008; 118: 2021\u0026ndash;24.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKojima M, Ishii G, Atsumi N, Fujii S, Saito N, Ochiai A. Immunohistochemical detection of CD133 expression in colorectal cancer: a clinicopathological study. Cancer Sci. 2008;99: 1578e1583.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHorst D, Kriegl L, Engel J, Kirchner T, Jung A. CD133 expression is an independent prognostic marker for low survival in colorectal cancer. Br J Cancer. 2008;99:1285e1289.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang K, Xu J, Zhang J, Huang J. Prognostic role of CD133 expression in colorectal cancer: a meta-analysis. BMC Cancer. 2012 Dec 5;12:573.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHuang R, Mo D Wu J, Ai H, Lu Y.CD133 expression correlates with clinicopathologic features and poor prognosis of colorectal cancer patients: An updated meta-analysis of 37 studies. Medicine (Baltimore). 2018 Jun;97(23):e10446.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKim ST, Sohn I, DO IG, et al. Transcriptome analysis of CD133-positive stem cells and prognostic value of survivin in colorectal cancer. Cancer Genomics Proteomics. 2014 Sep-Oct;11(5):259\u0026ndash;66.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eReggiani Bonetti L, Migaldi M, Caredda E, et al. Increased expression of CD133 is a strong predictor of poor outcome in stage I colorectal cancer patients. Scand J Gastroenterol. 2012 Oct;47(10):1211\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePitule P, Cedikova M, Daum O, et al. Immunohistochemical detection of cancer stem cell related markers CD44 and CD133 in metastatic colorectal cancer patients. Biomed Res Int. 2014;2014:432139.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHosseini S, Izadpanah A, Yarmohammadi H. Epidemiological changes in colorectal cancer in Shiraz, Iran: 1980\u0026ndash;2000. ANZ J Surg 2004; 74:547\u0026ndash;49.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAnsari R, Mahdavinia M, Sadjadi A, Nouraie M, Kamangar F, Bishehsari F, et al. Incidence and age distribution of colorectal cancer in Iran: results of a population-based cancer registry. Cancer Lett 18 2006;240:143\u0026ndash;47.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eForoutan M, Rahimi N, Tabatabaeifar M, Darvishi M, Hashemi M, Hosseinpanah F, et al. Clinical features of colorectal cancer in Iran: A 15-year review. Dig Dis 2008; 9:225\u0026ndash;27.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePahlavan PS, Jensen K. A short impact of epidemiological features of colorectal cancer in Iran. Tumori 2005; 91:291\u0026ndash;94.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHong I, Hong SW, Chang YG, Lee WY, Lee B, Kang YK, Kim YS, Paik IW, Lee H. Expression of the cancer stem cell markers CD44 and CD133 in colorectal cancer: an immunohistochemical staining analysis.Ann Coloproctol 2015; 31:84\u0026ndash;91.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHongo K, Kazama S, Sunami E, Tsuno NH, Takahashi K, Nagawa H, Kitayama J. Immunohistochemical detection of CD133 is associated with tumor regression grade after chemoradiotherapy in rectal cancer. Med Oncol 2012; 29:2849\u0026ndash;57.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKojima M, Ishii G, Atsumi N, Fujii S, Saito N, Ochiai A. Immunohistochemical detection of CD133 expression in colorectal cancer: a clinicopathological study. Cancer Sci 2008; 99:1578\u0026ndash;83.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePitule P, Cedikova M, Daum O, Vojtisek J, Vycital O, Hosek P, et al. Immunohistochemical detection of cancer stem cell related markers CD44 and CD133 in metastatic colorectal cancer patients. Biomed Res Int 2014;2014:432139.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen S, Song X, Chen Z, et al. CD133 expression and the prognosis of colorectal cancer: a systematic review and metaanalysis. PLoS One. 2013;8:e56380.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSchneider M, Huber J, Hadaschik B, Siegers GM, Fiebig H-H, Sch\u0026uuml;ler J. Characterization of colon cancer cells: a functional approach characterizing CD133 as a potential stem cell marker. BMC cancer. 2012;12(1):1\u0026ndash;11.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYang Z-L, Zheng Q, Yan J, Pan Y, Wang Z-G. Upregulated CD133 expression in tumorigenesis of colon cancer cells. World journal of gastroenterology: WJG. 2011;17(7):932.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRicci-Vitiani L, LombardiDG, Pilozzi E, et al. Identification and expansion of human colon-cancer-initiating cells.Nature 2007;445:111^5. \\\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHorst D, Kriegl L, Engel J, Kirchner T, Jung A. Prognostic significance of the cancer stem cell markers CD133, CD44, and CD166 in colorectal cancer. Cancer Invest 2009; 27:844\u0026ndash;50.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHuang R, Mo D, Wu J, Ai H, Lu Y. CD133 expression correlates with clinicopathologic features and poor prognosis of colorectal cancer patients: An updated meta-analysis of 37 studies. Medicine. 2018;97(23).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNosrati A, Naghshvar F, Maleki I, Salehi F. Cancer stem cells CD133 and CD24 in colorectal cancers n Northern Iran. Gastroenterol Hepatol Bed Bench 2016; 9:132\u0026ndash;9\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIinuma H, Watanabe T, Mimori K, et al. Clinical significance of circulating tumor cells, including cancer stem-like cells, in peripheral blood for recurrence and prognosis in patients with Dukes\u0026rsquo; stage B and C colorectal cancer. J Clin Oncol 2011;29:1547\u0026ndash;55.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJao SW, Chen SF, Lin YS, et al. Cytoplasmic CD133 expression is a reliable prognostic indicator of tumor regression after neoadjuvant concurrent chemoradiotherapy in patients with rectal cancer. Ann Surg Oncol 2012;19:3432\u0026ndash;40.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChoi D, Lee HW, Hur KY, Kim JJ, Park GS, Jang SH, et al. Cancer stem cell markers CD133 and CD24 correlate with invasiveness and differentiation in colorectal adenocarcinoma. World J Gastroenterol 2009;15:2258\u0026ndash;64.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLugli A, Iezzi G, Hostettler I, Muraro MG, Mele V, Tornillo L, et al. Prognostic impact of the expression of putative cancer stem cell markers CD133, CD166, CD44s, EpCAM, and ALDH1 in colorectal cancer. Br J Cancer 2010;103:382\u0026ndash;90.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhou F, Mu YD, Liang J, Liu ZX, Chen HS, Zhang JF. Expression and prognostic value of tumor stem cell markers ALDH1 and CD133 in colorectal carcinoma. Oncol Lett 2014;7:507\u0026ndash;12.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePark YY, An CH, Oh ST, Chang ED, Lee J. Expression of CD133 is associated with poor prognosis in stage II colorectal carcinoma. Medicine. 2019;98(32).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Carcinoma, Colon, Immunohistochemistry, Neoplastic stem cells, Rectum","lastPublishedDoi":"10.21203/rs.3.rs-1538147/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-1538147/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e Today, colon cancer is one of the most common types of gastrointestinal cancers CD133 as a known cancer stem cell marker has been found effective in cell proliferation and differentiation in various cancers, including colon. We aimed to investigate the relationship between the CD133 expression in colon cancer with prognostic factors and the survival rate of colon cancer patients by immunohistochemistry. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eMaterials and Methods: \u003c/strong\u003eFormalin- fixed-paraffinembedded (FFPE) tissues was taken from colon patients.\u0026nbsp;Histopathology examination was done using hematoxylin and eosin staining. \u0026nbsp;Immunohistochemistry was performed to determine CD133 expression. An association between CD133 expression and clinic pathological profile was then analyzed.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e There was a statistically significant difference between CD133 protein expression and sex, cancer stage and lymphatic invasion (p=0.044, p=0.131 and p=0.002, respectively). However, no significant correlation was identified between the CD133 expression and other factors such as: age of CRC patients (\u003cem\u003ep\u003c/em\u003e-value=0.267), tumor location (\u003cem\u003ep\u003c/em\u003e-value=0.494), tumor differentiation grade (\u003cem\u003ep\u003c/em\u003e-value=0.263), neural tissue invasion and 5 years’ survival (p value=0.054). \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConclusion: \u003c/strong\u003eCD133 is a useful predictive or prognostic biomarker for CRC in clinical assessment and may serve as a potential therapeutic target for CRC.\u003c/p\u003e","manuscriptTitle":"Expression and Prognostic Significance of Stem Cell Marker CD133 in Survival Rate of Colon Cancer Patients","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-04-18 14:22:13","doi":"10.21203/rs.3.rs-1538147/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"dfa38f47-1957-47d5-ad07-96c5b9614710","owner":[],"postedDate":"April 18th, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2022-04-18T14:22:14+00:00","versionOfRecord":[],"versionCreatedAt":"2022-04-18 14:22:13","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-1538147","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-1538147","identity":"rs-1538147","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}