HMGA2 rs968697 T>C Polymorphism is Associated With The Risk of Colorectal Cancer

HMGA2 rs968697 T>C Polymorphism is Associated With The Risk of Colorectal Cancer | 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 Original Article HMGA2 rs968697 T>C Polymorphism is Associated With The Risk of Colorectal Cancer Zhidan Li, Jiaojiao Yang, Shulong Zhang, Xiaoting Wang, Xueren Gao This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-159922/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: Recently, a genetic polymorphism (rs968697 T>C) in HMGA2 gene has been reported to be associated with hepatoblastoma risk. However, no studies reported the effect of the polymorphism on the risk of colorectal cancer (CRC). The study aimed to explore whether rs968697 polymorphism had a significant impact on CRC risk. Methods: A total of 500 CRC patients and 500 age and gender matched healthy individuals were genotyped by Sanger sequencing. Quantitative real-time PCR technology was used to detect the relative expression of HMGA2 gene in 30 pairs of primary CRC and adjacent non-cancerous tissues. Results: HMGA2 rs968697 polymorphism was significantly associated with CRC risk (CC vs. TT: OR=0.20, 95%CI=0.06-0.70, P=0.01; (CC+CT) vs. TT: OR=0.71, 95%CI=0.53-0.96, P=0.02; CC vs. (CT+TT): OR=0.21, 95%CI=0.06-0.73, P=0.01; C vs. T: OR=0.67, 95%CI=0.51-0.89, P<0.01). The analysis based on tumor stage indicated that HMGA2 rs968697 polymorphism was significantly associated with CRC tumor stage. In addition, the genotype-tissue expression showed that the rs968697 polymorphism was related to HMGA2 gene expression. The in silico analysis showed that rs968697 polymorphism located in the promoter region of HMGA2 gene could affect the binding of transcription factors. Conclusion: Our study suggested that HMGA2 rs968697 polymorphism was associated with CRC risk and might serve as a reliable biomarker to detect CRC risk. Running head: HMGA2 rs968697 T>C polymorphism and colorectal cancer General Biochemistry Biotechnology and Bioengineering HMGA2 colorectal cancer risk polymorphism Figures Figure 1 1. Introduction High mobility group AT-hook 2 ( HMGA2 ) gene is located on chromosome12q14.3, and encodes an architectural transcription factor that belongs to the non-histone chromosomal high mobility group protein family. The protein can act as an oncogenic protein closely linked to aggressive tumor behavior and poor clinical outcomes [ 1 – 3 ]. Thereinto the promoting role of HMGA2 in the occurrence and development of colorectal cancer (CRC) has been reported [ 4 – 5 ]. For instance, HMGA2 over-expression was associated not only with metastasis but also with reduced survival rates of CRC patients [ 4 ]. HMGA2 could promote intestinal tumorigenesis by facilitating MDM2-mediated ubiquitination and degradation of p53 [ 5 ]. In recent years, a case-control study also investigated the association of a genetic polymorphism (rs968697 T > C) in the cancer-related gene with cancer risk, and found that the polymorphism was significantly associated with hepatoblastoma risk. In view of the significance of rs968697 polymorphism and the critical role of HMGA2 gene in CRC, we speculated that HMGA2 rs968697 polymorphism might be involved in CRC risk. However, no studies reported the effect of the polymorphism on CRC risk. Thus, a case-control study was conducted to assess the strength of the effect. 2. Methods And Materials 2.1 Peripheral blood sample collection Peripheral blood samples of 500 CRC patients and 500 age and gender matched healthy individuals were collected from Xuhui District Central Hospital of Shanghai between October 2017 and January 2021. The mean age of CRC patients and healthy individuals was 59.1 ± 7.4 and 59.4 ± 7.7 years, respectively. The ratio of male and female was 53:47 in both CRC patients and healthy individuals. Diagnosis of all patients was histopathologically confirmed. Healthy individuals were cancer-free individuals living in the same residential area and seeking for routine physical examination. This study protocol was approved by the Institutional Review Board of the Hospital. All enrolled subjects provided written informed consent in the study. 2.2 Genotyping Genomic DNA was extracted from peripheral blood samples by using the TIANamp genomic DNA Kit (Tiangen). The concentration and quality of genomic DNA were determined by using a NanoDrop spectrophotometer (Thermo Fisher Scientific). Genotyping was performed by Sanger sequencing. 2.3 The relationship between rs968697 T > C polymorphism and HMGA2 gene expression Total RNA was extracted from 30 pairs of primary CRC and adjacent non-cancerous tissues by using the RNAsimple total RNA kit (Tiangen) according to the manufacturer’s instructions. The cDNA was synthesized using ReverTra Ace qPCR RT Kit (TOYOBO). Quantitative Real-time PCR was performed using Roche FastStart Universal SYBR Green Master (Rox). The expression level of HMGA2 gene was normalized to the internal control GAPDH. The 2 −ΔΔCt method was used to determine HMGA2 gene expression. The specific primer pairs for HMGA2 and GAPDH were 5′-CAAGTTGTTCAGAAGAAGCC-3′ (forward), 5′-GGCAATACAG AATAAGTGGTC-3′(reverse); and 5′-GTCTCCTCTGACTTCAACA-3′ (forward), 5′-TGAGGG TCTCTCTCTTCCT-3′ (reverse), respectively. 2.4 Bioinformatics analysis RegulomeDB ( www.regulomedb.org/regulome-search/ ) is a database annotating SNPs with known and predicted regulatory elements in the intergenic regions of the human genome [ 6 ]. SNPinfo ( https://snpinfo.niehs.nih.gov/snpinfo/snpfunc.html ) is a web-based tool integrating genome-wide association studies and candidate gene information into functional SNP selection for genetic association studies [ 7 ]. In this study, RegulomeDB and SNPinfo were used to explore the latent function of HMGA2 rs968697 polymorphism. For RegulomeDB database, POLR2A ChIP-Seq data from human colon cancer cell (HCT116) was analyzed. 2.5 Statistical analysis In control group, Hardy-Weinberg equilibrium (HWE) of the genotype distribution was analyzed using the chi-square goodness-of-fit test. The association between HMGA2 rs968697 polymorphism and CRC risk was evaluated using logistic regression analysis. The association between HMGA2 rs968697 polymorphism and CRC tumor stage was analyzed using the chi-square test. The expression levels of HMGA2 gene were compared among different genotype groups by using the one-way ANOVA. Statistical analyses were performed using SPSS 22.0 software. P value less than 0.05 was considered statistically significant. 3. Results 3.1 HMGA2 rs968697 was significantly associated with CRC risk As shown in Table 1 , the genotype distribution of HMGA2 rs968697 polymorphism was 371/115/14 (TT/CT/CC) in control group, which conformed to HWE (P = 0.17). In CRC patients, the genotype distribution of HMGA2 rs968697 polymorphism was 400/97/3 (TT/CT/CC). The comparison results showed that HMGA2 rs968697 polymorphism was significantly associated with CRC risk. The individuals carrying rs968697 CC genotype had a decreased risk of CRC compared with those carrying the TT genotype (OR = 0.20; 95%CI = 0.06–0.70; P = 0.01). The individuals carrying rs968697 CC and CT genotype had a decreased risk of CRC compared with those carrying the TT genotype (OR = 0.71; 95%CI = 0.53–0.96; P = 0.02). The individuals carrying rs968697 CC genotype had a decreased risk of CRC compared with those carrying the CT and TT genotype (OR = 0.21; 95%CI = 0.06–0.73; P = 0.01). The individuals carrying rs968697 C allele had a decreased risk of CRC compared with those carrying the T allele (OR = 0.67; 95%CI = 0.51–0.89; P < 0.01). Furthermore, the analysis based on tumor stage indicated that rs968697 polymorphism was significantly associated with CRC tumor stage (Table 2 ). Table 1 The association of HMGA2 rs968697 polymorphism with CRC risk Genotype/allele Cases (N = 500) Controls (N = 500) Comparison a OR (95%CI) a P TT 400 (80.0%) 371 (74.2%) CT vs. TT 0.77 (0.57–1.05) 0.10 CT 97 (19.4%) 115 (23.0%) CC vs. TT 0.20 (0.06–0.70) 0.01 CC 3 (0.6%) 14 (2.8%) (CC + CT) vs. TT 0.71 (0.53–0.96) 0.02 T 897 (89.7%) 857 (85.7%) CC vs. (CT + TT) 0.21 (0.06–0.73) 0.01 C 103 (10.3%) 143 (14.3%) C vs. T 0.67 (0.51–0.89) < 0.01 a : adjusted for gender and age; OR: Odds ratio; CI: Confidence interval. Table 2 The association of HMGA2 rs968697 polymorphism with CRC tumor stage Genotype/allele I + II (N = 265) III + IV (N = 235) P TT 198 (74.7%) 202 (86.0%) 0.01 CT 65 (24.5%) 32 (13.6%) CC 2 (0.8%) 1 (0.4%) T 461 (87.0%) 436 (92.8%) < 0.01 C 69 (13.0%) 34 (7.2%) 3.2 Rs968697 polymorphism was related to HMGA2 gene expression As shown in Fig. 1 , the genotype-tissue expression showed that rs968697 polymorphism was related to the expression of HMGA2 gene. HMGA2 gene had a higher expression level in CRC and adjacent non-cancerous tissues with rs968697 TT genotype than in CRC and adjacent non-cancerous tissues with rs968697 CC genotype. 3.3 Rs968697 polymorphism affected the binding of transcription factors The in silico analysis showed that rs968697 polymorphism located in the promoter region of HMGA2 gene affected the binding of transcription factors, such as DBP, CDPCR3, POLR2A and TAXCREB (Table 3 ). Table 3 Bioinformatics analysis for the latent function of HMGA2 rs968697 polymorphism. SNP Predictive tools Transcription factors rs968697 SNPinfo ATF6, DBP, CDPCR3, DR3, NRSF, PAX8, PPARA, SZF11, TAXCREB RegulomeDB POLR2A 4. Discussion As one of the most common cancers, CRC was estimated to cause over 1.8 million new cases and 881,000 deaths in 2018 [ 8 ]. Although the molecular etiology of CRC is still unknown, its onset is significantly associated with genetic variants. Single nueleotide polymorphism (SNP) is one of the most common genetic variants and involves altered cancer risk [ 9 – 11 ]. In the current study, we conducted a case-control study investigating the association between HMGA2 rs968697 polymorphism and CRC risk, and found that rs968697 C allele could reduce CRC risk compared with the T allele. To the best of our knowledge, this is the first epidemiological study exploring the association of HMGA2 rs968697 polymorphism with CRC risk in the Chinese population. Furthermore, we also found that rs968697 polymorphism was related to the expression of HMGA2 gene in CRC and adjacent non-cancerous tissues. Bioinformatics analysis showed that the rs968697 polymorphism had an effect on the binding of transcription factors to the promoter of HMGA2 gene, which provided a possible molecular explanation for the effect of the genetic polymorphism on CRC risk. However, the detailed mechanism still needs to be verified by further well-designed experiment. It is worthy of note that the current sample size is relative small, which may not obtain sufficient statistical power. Therefore, further replication studies are necessary to fully establish the association between HMGA2 rs968697 polymorphism and CRC risk. 5. Conclusion Our molecular epidemiological findings demonstrated a significant association of HMGA2 rs968697 polymorphism with CRC risk. The rs968697 polymorphism may serve as a potential biomarker for CRC risk. Declarations Ethical approval and consent to participate The study was approved by the Ethics Committee of Yancheng Teachers’ University, and all participants provided written informed consent. Authors contributions and consent to publish ZL drafted the manuscript. SZ collected clinical samples. JY and XW were responsible for the figures and tables. XG critically revised the manuscript. All authors approved the final manuscript. Funding This work was supported by scientific research project of Shanghai Municipal Health Commission (No. 201940389). Competing Interests The authors declare no conflict of interest. Availability of data and materials The data and materials used or analysed during the current study are available on request to the corresponding author. References Sun, J., Sun, B., Sun, R., Zhu, D., Zhao, X., Zhang, Y., Dong, X., Che, N., Li, J., Liu, F., Zhao, N., Wang, Y., & Zhang, D. (2017) HMGA2 promotes vasculogenic mimicry and tumor aggressiveness by upregulating Twist1 in gastric carcinoma. Sci Rep 7 , 2229. Zhang, H., Tang, Z., Deng, C., He, Y., Wu, F., Liu, O., & Hu, C. (2017) HMGA2 is associated with the aggressiveness of tongue squamous cell carcinoma. Oral Dis 23 , 255–264. Wu, J., Zhang, S., Shan, J., Hu, Z., Liu, X., Chen, L., Ren, X., Yao, L., Sheng, H., Li, L., Ann, D., Yen, Y., Wang, J., & Wang, X. (2016) Elevated HMGA2 expression is associated with cancer aggressiveness and predicts poor outcome in breast cancer. Cancer Lett 376 , 284–292. Wang, X., Liu, X., Li, A. Y., Chen, L., Lai, L., Lin, H. H., Hu, S., Yao, L., Peng, J., Loera, S., Xue, L., Zhou, B., Zhou, L., Zheng, S., Chu, P., Zhang, S., Ann, D. K., & Yen, Y. (2011) Overexpression of HMGA2 promotes metastasis and impacts survival of colorectal cancers. Clin Cancer Res 17 , 2570–2580. Wang, Y., Hu, L., Wang, J., Li, X., Sahengbieke, S., Wu, J., & Lai, M. (2018) HMGA2 promotes intestinal tumorigenesis by facilitating MDM2-mediated ubiquitination and degradation of p53. J Pathol 246 , 508–518. Boyle, A. P., Hong, E. L., Hariharan, M., Cheng, Y., Schaub, M. A., Kasowski, M., Karczewski, K. J., Park, J., Hitz, B. C., Weng, S., Cherry, J. M., & Snyder, M. (2012) Annotation of functional variation in personal genomes using RegulomeDB. Genome Res 22 , 1790–1797. Xu, Z., & Taylor, J. A. (2009) SNPinfo: integrating GWAS and candidate gene information into functional SNP selection for genetic association studies. Nucleic Acids Res 37 (Web Server issue): W600-W605. Bray, F., Ferlay, J., Soerjomataram, I., Siegel, R. L., Torre, L. A., & Jemal, A. (2018) Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 68 , 394–424. Ni, W., Wang, X., Sun, Y., & Gao, X. (2020) Meta-analysis of the association between MALAT1 rs619586 A>G polymorphism and cancer risk. J Int Med Res 48 :300060520941969. Gao, X., Zhu, Z., & Zhang, S. (2018) miR-146a rs2910164 polymorphism and the risk of colorectal cancer in Chinese population. J Cancer Res Ther 14 (Supplement): S97-S99. Xiao, Y., Dong, Z., Zhu, J., You, J., & Fan, J. (2019) Association between ACE A240T polymorphism and cancer risk: a meta-analysis. J Int Med Res 47 , 5917–5925. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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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-159922","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Original Article","associatedPublications":[],"authors":[{"id":9308275,"identity":"d136021f-fa7a-44d2-8e02-e8861c366594","order_by":0,"name":"Zhidan Li","email":"","orcid":"","institution":"Yancheng Teachers University","correspondingAuthor":false,"prefix":"","firstName":"Zhidan","middleName":"","lastName":"Li","suffix":""},{"id":9308276,"identity":"43891b5c-a3fe-4e2e-9d0c-b5cfeef709b3","order_by":1,"name":"Jiaojiao Yang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAq0lEQVRIiWNgGAWjYFCCA4kPoCwDorUkw5QSrYWBTYI0LQYHDzyr/Nm2LbGBvXmbBEPNHSK0HDiQdpvnzO3EBp5jZRIMx54RqYWhAqhFIsdMgrHhMHFaCn8YALXIvyFBCwMP2BYeIrVIHjiQLA30i3EbT1qxRcIxIrTw3TiT+PFn223ZfvbDG298qCFCi8KNMwlgBhuISCCsgYFBvr/9ADHqRsEoGAWjYCQDABGnQU0GWrYKAAAAAElFTkSuQmCC","orcid":"","institution":"Shanxi Medical University","correspondingAuthor":true,"prefix":"","firstName":"Jiaojiao","middleName":"","lastName":"Yang","suffix":""},{"id":9308277,"identity":"f2d8a04c-fddf-4ac8-8d17-88694155f89b","order_by":2,"name":"Shulong Zhang","email":"","orcid":"","institution":"Shanghai Xuhui Central Hospital","correspondingAuthor":false,"prefix":"","firstName":"Shulong","middleName":"","lastName":"Zhang","suffix":""},{"id":9308278,"identity":"4ccf9d6b-60a9-4e16-8b08-055459856909","order_by":3,"name":"Xiaoting Wang","email":"","orcid":"","institution":"Shanghai Xuhui Central Hospital","correspondingAuthor":false,"prefix":"","firstName":"Xiaoting","middleName":"","lastName":"Wang","suffix":""},{"id":9308279,"identity":"a8530142-afd6-4e4d-9719-f4dcda856128","order_by":4,"name":"Xueren Gao","email":"","orcid":"","institution":"Yancheng Teachers University","correspondingAuthor":false,"prefix":"","firstName":"Xueren","middleName":"","lastName":"Gao","suffix":""}],"badges":[],"createdAt":"2021-01-27 18:02:56","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-159922/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-159922/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":5554320,"identity":"d1cc05a0-35d8-48b7-97bd-3d97e1acfed7","added_by":"auto","created_at":"2021-02-02 19:42:24","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":34782,"visible":true,"origin":"","legend":"The relationship between rs968697 polymorphism and the expression of HMGA2 gene in 30 pairs of primary CRC and adjacent non-cancerous tissues (A: adjacent non-cancerous tissues; B: CRC tissues; *:P\u003c0.05; **:P\u003c0.01).","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-159922/v1/0ec09908ab964b9cc3270aa8.png"},{"id":13654412,"identity":"48eab0e9-81a4-45ce-966b-6aefee3a6442","added_by":"auto","created_at":"2021-09-17 09:57:32","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":427102,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-159922/v1/b0be7fba-6131-41fe-88f2-bf62d706d4e5.pdf"}],"financialInterests":"","formattedTitle":"\u003cp\u003eHMGA2 rs968697 T\u0026gt;C Polymorphism is Associated With The Risk of Colorectal Cancer\u003c/p\u003e","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eHigh mobility group AT-hook 2 (\u003cem\u003eHMGA2\u003c/em\u003e) gene is located on chromosome12q14.3, and encodes an architectural transcription factor that belongs to the non-histone chromosomal high mobility group protein family. The protein can act as an oncogenic protein closely linked to aggressive tumor behavior and poor clinical outcomes [\u003cspan additionalcitationids=\"CR2\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Thereinto the promoting role of HMGA2 in the occurrence and development of colorectal cancer (CRC) has been reported [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. For instance, HMGA2 over-expression was associated not only with metastasis but also with reduced survival rates of CRC patients [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. HMGA2 could promote intestinal tumorigenesis by facilitating MDM2-mediated ubiquitination and degradation of p53 [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. In recent years, a case-control study also investigated the association of a genetic polymorphism (rs968697 T\u0026thinsp;\u0026gt;\u0026thinsp;C) in the cancer-related gene with cancer risk, and found that the polymorphism was significantly associated with hepatoblastoma risk.\u003c/p\u003e\u003cp\u003eIn view of the significance of rs968697 polymorphism and the critical role of \u003cem\u003eHMGA2\u003c/em\u003e gene in CRC, we speculated that \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism might be involved in CRC risk. However, no studies reported the effect of the polymorphism on CRC risk. Thus, a case-control study was conducted to assess the strength of the effect.\u003c/p\u003e"},{"header":"2. Methods And Materials","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1 Peripheral blood sample collection\u003c/h2\u003e\u003cp\u003ePeripheral blood samples of 500 CRC patients and 500 age and gender matched healthy individuals were collected from Xuhui District Central Hospital of Shanghai between October 2017 and January 2021. The mean age of CRC patients and healthy individuals was 59.1\u0026thinsp;\u0026plusmn;\u0026thinsp;7.4 and 59.4\u0026thinsp;\u0026plusmn;\u0026thinsp;7.7 years, respectively. The ratio of male and female was 53:47 in both CRC patients and healthy individuals. Diagnosis of all patients was histopathologically confirmed. Healthy individuals were cancer-free individuals living in the same residential area and seeking for routine physical examination. This study protocol was approved by the Institutional Review Board of the Hospital. All enrolled subjects provided written informed consent in the study.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2 Genotyping\u003c/h2\u003e\u003cp\u003eGenomic DNA was extracted from peripheral blood samples by using the TIANamp genomic DNA Kit (Tiangen). The concentration and quality of genomic DNA were determined by using a NanoDrop spectrophotometer (Thermo Fisher Scientific). Genotyping was performed by Sanger sequencing.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e2.3 The relationship between rs968697 T\u0026thinsp;\u0026gt;\u0026thinsp;C polymorphism and \u003cem\u003eHMGA2\u003c/em\u003e gene expression\u003c/h2\u003e\u003cp\u003e Total RNA was extracted from 30 pairs of primary CRC and adjacent non-cancerous tissues by using the RNAsimple total RNA kit (Tiangen) according to the manufacturer\u0026rsquo;s instructions. The cDNA was synthesized using ReverTra Ace qPCR RT Kit (TOYOBO). Quantitative Real-time PCR was performed using Roche FastStart Universal SYBR Green Master (Rox). The expression level of \u003cem\u003eHMGA2\u003c/em\u003e gene was normalized to the internal control GAPDH. The 2\u003csup\u003e\u0026minus;ΔΔCt\u003c/sup\u003e method was used to determine \u003cem\u003eHMGA2\u003c/em\u003e gene expression. The specific primer pairs for HMGA2 and GAPDH were 5\u0026prime;-CAAGTTGTTCAGAAGAAGCC-3\u0026prime; (forward), 5\u0026prime;-GGCAATACAG AATAAGTGGTC-3\u0026prime;(reverse); and 5\u0026prime;-GTCTCCTCTGACTTCAACA-3\u0026prime; (forward), 5\u0026prime;-TGAGGG TCTCTCTCTTCCT-3\u0026prime; (reverse), respectively.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003e2.4 Bioinformatics analysis\u003c/h2\u003e\u003cp\u003eRegulomeDB (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e\u003ca href=\"http://www.regulomedb.org/regulome-search/\" target=\"_blank\"\u003ewww.regulomedb.org/regulome-search/\u003c/a\u003e\u003c/span\u003e\u003c/span\u003e) is a database annotating SNPs with known and predicted regulatory elements in the intergenic regions of the human genome [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. SNPinfo (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://snpinfo.niehs.nih.gov/snpinfo/snpfunc.html\u003c/span\u003e\u003c/span\u003e) is a web-based tool integrating genome-wide association studies and candidate gene information into functional SNP selection for genetic association studies [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. In this study, RegulomeDB and SNPinfo were used to explore the latent function of HMGA2 rs968697 polymorphism. For RegulomeDB database, POLR2A ChIP-Seq data from human colon cancer cell (HCT116) was analyzed.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003e2.5 Statistical analysis\u003c/h2\u003e\u003cp\u003eIn control group, Hardy-Weinberg equilibrium (HWE) of the genotype distribution was analyzed using the chi-square goodness-of-fit test. The association between \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism and CRC risk was evaluated using logistic regression analysis. The association between \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism and CRC tumor stage was analyzed using the chi-square test. The expression levels of \u003cem\u003eHMGA2\u003c/em\u003e gene were compared among different genotype groups by using the one-way ANOVA. Statistical analyses were performed using SPSS 22.0 software. P value less than 0.05 was considered statistically significant.\u003c/p\u003e\u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\n\u003ch2\u003e3.1 \u003cem\u003eHMGA2\u003c/em\u003e rs968697 was significantly associated with CRC risk\u003c/h2\u003e\n\u003cp\u003eAs shown in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e, the genotype distribution of \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism was 371/115/14 (TT/CT/CC) in control group, which conformed to HWE (P\u0026thinsp;=\u0026thinsp;0.17). In CRC patients, the genotype distribution of \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism was 400/97/3 (TT/CT/CC). The comparison results showed that \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism was significantly associated with CRC risk. The individuals carrying rs968697 CC genotype had a decreased risk of CRC compared with those carrying the TT genotype (OR\u0026thinsp;=\u0026thinsp;0.20; 95%CI\u0026thinsp;=\u0026thinsp;0.06\u0026ndash;0.70; P\u0026thinsp;=\u0026thinsp;0.01). The individuals carrying rs968697 CC and CT genotype had a decreased risk of CRC compared with those carrying the TT genotype (OR\u0026thinsp;=\u0026thinsp;0.71; 95%CI\u0026thinsp;=\u0026thinsp;0.53\u0026ndash;0.96; P\u0026thinsp;=\u0026thinsp;0.02). The individuals carrying rs968697 CC genotype had a decreased risk of CRC compared with those carrying the CT and TT genotype (OR\u0026thinsp;=\u0026thinsp;0.21; 95%CI\u0026thinsp;=\u0026thinsp;0.06\u0026ndash;0.73; P\u0026thinsp;=\u0026thinsp;0.01). The individuals carrying rs968697 C allele had a decreased risk of CRC compared with those carrying the T allele (OR\u0026thinsp;=\u0026thinsp;0.67; 95%CI\u0026thinsp;=\u0026thinsp;0.51\u0026ndash;0.89; P\u0026thinsp;\u0026lt;\u0026thinsp;0.01). Furthermore, the analysis based on tumor stage indicated that rs968697 polymorphism was significantly associated with CRC tumor stage (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab1\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eThe association of \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism with CRC risk\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eGenotype/allele\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eCases\u003c/p\u003e\n\u003cp\u003e(N\u0026thinsp;=\u0026thinsp;500)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eControls\u003c/p\u003e\n\u003cp\u003e(N\u0026thinsp;=\u0026thinsp;500)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eComparison\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003csup\u003ea\u003c/sup\u003eOR (95%CI)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003csup\u003ea\u003c/sup\u003eP\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e400 (80.0%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e371 (74.2%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCT vs. TT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.77 (0.57\u0026ndash;1.05)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.10\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e97 (19.4%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e115 (23.0%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCC vs. TT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.20 (0.06\u0026ndash;0.70)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.01\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3 (0.6%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e14 (2.8%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(CC\u0026thinsp;+\u0026thinsp;CT) vs. TT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.71 (0.53\u0026ndash;0.96)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.02\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e897 (89.7%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e857 (85.7%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCC vs. (CT\u0026thinsp;+\u0026thinsp;TT)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.21 (0.06\u0026ndash;0.73)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.01\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e103 (10.3%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e143 (14.3%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC vs. T\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.67 (0.51\u0026ndash;0.89)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.01\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003ctfoot\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"6\"\u003e\u003csup\u003ea\u003c/sup\u003e: adjusted for gender and age; OR: Odds ratio; CI: Confidence interval.\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tfoot\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab2\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eThe association of \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism with CRC tumor stage\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eGenotype/allele\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eI\u0026thinsp;+\u0026thinsp;II\u003c/p\u003e\n\u003cp\u003e(N\u0026thinsp;=\u0026thinsp;265)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eIII\u0026thinsp;+\u0026thinsp;IV (N\u0026thinsp;=\u0026thinsp;235)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eP\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e198 (74.7%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e202 (86.0%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"3\" align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.01\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e65 (24.5%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e32 (13.6%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2 (0.8%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1 (0.4%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e461 (87.0%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e436 (92.8%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.01\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e69 (13.0%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e34 (7.2%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\n\u003ch2\u003e3.2 Rs968697 polymorphism was related to \u003cem\u003eHMGA2\u003c/em\u003e gene expression\u003c/h2\u003e\n\u003cp\u003eAs shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e, the genotype-tissue expression showed that rs968697 polymorphism was related to the expression of \u003cem\u003eHMGA2\u003c/em\u003e gene. \u003cem\u003eHMGA2\u003c/em\u003e gene had a higher expression level in CRC and adjacent non-cancerous tissues with rs968697 TT genotype than in CRC and adjacent non-cancerous tissues with rs968697 CC genotype.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n\u003ch2\u003e3.3 Rs968697 polymorphism affected the binding of transcription factors\u003c/h2\u003e\n\u003cp\u003eThe in silico analysis showed that rs968697 polymorphism located in the promoter region of \u003cem\u003eHMGA2\u003c/em\u003e gene affected the binding of transcription factors, such as DBP, CDPCR3, POLR2A and TAXCREB (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab3\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eBioinformatics analysis for the latent function of \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eSNP\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePredictive tools\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eTranscription factors\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003ers968697\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSNPinfo\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eATF6, DBP, CDPCR3, DR3, NRSF, PAX8, PPARA, SZF11, TAXCREB\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eRegulomeDB\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePOLR2A\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eAs one of the most common cancers, CRC was estimated to cause over 1.8\u0026nbsp;million new cases and 881,000 deaths in 2018 [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Although the molecular etiology of CRC is still unknown, its onset is significantly associated with genetic variants. Single nueleotide polymorphism (SNP) is one of the most common genetic variants and involves altered cancer risk [\u003cspan additionalcitationids=\"CR10\" citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. In the current study, we conducted a case-control study investigating the association between \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism and CRC risk, and found that rs968697 C allele could reduce CRC risk compared with the T allele. To the best of our knowledge, this is the first epidemiological study exploring the association of \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism with CRC risk in the Chinese population. Furthermore, we also found that rs968697 polymorphism was related to the expression of HMGA2 gene in CRC and adjacent non-cancerous tissues. Bioinformatics analysis showed that the rs968697 polymorphism had an effect on the binding of transcription factors to the promoter of \u003cem\u003eHMGA2\u003c/em\u003e gene, which provided a possible molecular explanation for the effect of the genetic polymorphism on CRC risk. However, the detailed mechanism still needs to be verified by further well-designed experiment. It is worthy of note that the current sample size is relative small, which may not obtain sufficient statistical power. Therefore, further replication studies are necessary to fully establish the association between \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism and CRC risk.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eOur molecular epidemiological findings demonstrated a significant association of \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism with CRC risk. The rs968697 polymorphism may serve as a potential biomarker for CRC risk.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003e Ethical approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study was approved by the Ethics Committee of Yancheng Teachers\u0026rsquo; University, and all participants provided written informed consent.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors contributions and consent to publish\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eZL drafted the manuscript. SZ collected clinical samples. JY and XW were responsible for the figures and tables. XG critically revised the manuscript. All authors approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by scientific research project of Shanghai Municipal Health Commission (No. 201940389).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data and materials used or analysed during the current study are available on request to the corresponding author.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eSun, J., Sun, B., Sun, R., Zhu, D., Zhao, X., Zhang, Y., Dong, X., Che, N., Li, J., Liu, F., Zhao, N., Wang, Y., \u0026amp; Zhang, D. (2017) HMGA2 promotes vasculogenic mimicry and tumor aggressiveness by upregulating Twist1 in gastric carcinoma. \u003cem\u003eSci Rep\u003c/em\u003e \u003cstrong\u003e7\u003c/strong\u003e, 2229.\u003c/li\u003e\n\u003cli\u003eZhang, H., Tang, Z., Deng, C., He, Y., Wu, F., Liu, O., \u0026amp; Hu, C. (2017) HMGA2 is associated with the aggressiveness of tongue squamous cell carcinoma.\u003cem\u003e Oral Dis \u003c/em\u003e\u003cstrong\u003e23\u003c/strong\u003e, 255\u0026ndash;264.\u0026nbsp;\u0026nbsp;\u003c/li\u003e\n\u003cli\u003eWu, J., Zhang, S., Shan, J., Hu, Z., Liu, X., Chen, L., Ren, X., Yao, L., Sheng, H., Li, L., Ann, D., Yen, Y., Wang, J., \u0026amp; Wang, X. (2016) Elevated HMGA2 expression is associated with cancer aggressiveness and predicts poor outcome in breast cancer.\u003cem\u003e Cancer Lett\u003c/em\u003e \u003cstrong\u003e376\u003c/strong\u003e, 284\u0026ndash;292.\u003c/li\u003e\n\u003cli\u003eWang, X., Liu, X., Li, A. Y., Chen, L., Lai, L., Lin, H. H., Hu, S., Yao, L., Peng, J., Loera, S., Xue, L., Zhou, B., Zhou, L., Zheng, S., Chu, P., Zhang, S., Ann, D. K., \u0026amp; Yen, Y. (2011) Overexpression of HMGA2 promotes metastasis and impacts survival of colorectal cancers. \u003cem\u003eClin Cancer Res\u003c/em\u003e\u003cstrong\u003e 17\u003c/strong\u003e, 2570\u0026ndash;2580. \u0026nbsp;\u0026nbsp;\u003c/li\u003e\n\u003cli\u003eWang, Y., Hu, L., Wang, J., Li, X., Sahengbieke, S., Wu, J., \u0026amp; Lai, M. (2018) HMGA2 promotes intestinal tumorigenesis by facilitating MDM2-mediated ubiquitination and degradation of p53. \u003cem\u003eJ Pathol \u003c/em\u003e\u003cstrong\u003e246\u003c/strong\u003e, 508\u0026ndash;518.\u003c/li\u003e\n\u003cli\u003eBoyle, A. P., Hong, E. L., Hariharan, M., Cheng, Y., Schaub, M. A., Kasowski, M., Karczewski, K. J., Park, J., Hitz, B. C., Weng, S., Cherry, J. M., \u0026amp; Snyder, M. (2012) Annotation of functional variation in personal genomes using RegulomeDB. \u003cem\u003eGenome Res \u003c/em\u003e\u003cstrong\u003e22\u003c/strong\u003e, 1790\u0026ndash;1797.\u003c/li\u003e\n\u003cli\u003eXu, Z., \u0026amp; Taylor, J. A. (2009) SNPinfo: integrating GWAS and candidate gene information into functional SNP selection for genetic association studies. \u003cem\u003eNucleic Acids Res \u003c/em\u003e\u003cstrong\u003e37 \u003c/strong\u003e(Web Server issue): W600-W605.\u003c/li\u003e\n\u003cli\u003eBray, F., Ferlay, J., Soerjomataram, I., Siegel, R. L., Torre, L. A., \u0026amp; Jemal, A. (2018) Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. \u003cem\u003eCA Cancer J Clin\u003c/em\u003e \u003cstrong\u003e68\u003c/strong\u003e, 394\u0026ndash;424.\u003c/li\u003e\n\u003cli\u003eNi, W., Wang, X., Sun, Y., \u0026amp; Gao, X. (2020) Meta-analysis of the association between MALAT1 rs619586 A\u0026gt;G polymorphism and cancer risk. \u003cem\u003eJ Int Med Res\u003c/em\u003e \u003cstrong\u003e48\u003c/strong\u003e:300060520941969.\u003c/li\u003e\n\u003cli\u003eGao, X., Zhu, Z., \u0026amp; Zhang, S. (2018) miR-146a rs2910164 polymorphism and the risk of colorectal cancer in Chinese population. \u003cem\u003eJ Cancer Res Ther \u003c/em\u003e\u003cstrong\u003e14 \u003c/strong\u003e(Supplement): S97-S99.\u003c/li\u003e\n\u003cli\u003eXiao, Y., Dong, Z., Zhu, J., You, J., \u0026amp; Fan, J. (2019) Association between ACE A240T polymorphism and cancer risk: a meta-analysis. \u003cem\u003eJ Int Med Res\u003c/em\u003e \u003cstrong\u003e47\u003c/strong\u003e, 5917\u0026ndash;5925.\u003c/li\u003e\n\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":"HMGA2, colorectal cancer, risk, polymorphism","lastPublishedDoi":"10.21203/rs.3.rs-159922/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-159922/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eRecently, a genetic polymorphism (rs968697 T\u0026gt;C) in \u003cem\u003eHMGA2 \u003c/em\u003egene has been reported to be associated with hepatoblastoma risk. However, no\u0026nbsp;studies\u0026nbsp;reported\u0026nbsp;the effect of the polymorphism on the risk of colorectal cancer (CRC). The study aimed to explore whether rs968697 polymorphism had\u0026nbsp;a\u0026nbsp;significant impact on CRC risk. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e A total of 500 CRC patients and 500 age and gender matched healthy individuals were genotyped by Sanger sequencing. Quantitative real-time PCR technology was used to detect the relative expression of \u003cem\u003eHMGA2\u003c/em\u003e gene in 30 pairs of primary CRC and adjacent non-cancerous tissues.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e\u003cem\u003e HMGA2\u003c/em\u003e rs968697 polymorphism was significantly associated with CRC risk (CC vs. TT: OR=0.20, 95%CI=0.06-0.70, P=0.01; (CC+CT) vs. TT: OR=0.71, 95%CI=0.53-0.96, P=0.02; CC vs. (CT+TT): OR=0.21, 95%CI=0.06-0.73, P=0.01; C vs. T: OR=0.67, 95%CI=0.51-0.89, P\u0026lt;0.01). The analysis based on tumor stage indicated that \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism was significantly associated with CRC tumor stage. In addition, the genotype-tissue expression showed that the rs968697 polymorphism was related to \u003cem\u003eHMGA2\u003c/em\u003e gene expression. The in silico analysis showed that rs968697 polymorphism located in the promoter region of \u003cem\u003eHMGA2\u003c/em\u003e gene could affect the binding of transcription factors. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConclusion: \u003c/strong\u003eOur study suggested that \u003cem\u003eHMGA2\u003c/em\u003e rs968697 polymorphism was associated with CRC risk and might serve as a reliable biomarker to detect CRC risk. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eRunning head:\u003cem\u003e \u003c/em\u003e\u003c/strong\u003e\u003cem\u003eHMGA2\u003c/em\u003e rs968697 T\u0026gt;C polymorphism and colorectal cancer\u003c/p\u003e","manuscriptTitle":"HMGA2 rs968697 T\u0026gt;C Polymorphism is Associated With The Risk of Colorectal Cancer","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-02-02 19:42:22","doi":"10.21203/rs.3.rs-159922/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":"207305e9-73c1-4c08-af8b-7ab2c8c585ae","owner":[],"postedDate":"February 2nd, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":2185633,"name":"General Biochemistry"},{"id":2185634,"name":"Biotechnology and Bioengineering"}],"tags":[],"updatedAt":"2021-02-02T19:42:23+00:00","versionOfRecord":[],"versionCreatedAt":"2021-02-02 19:42:22","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-159922","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-159922","identity":"rs-159922","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}