Mast cell density in gastric cancer and its relation to aggressive behavior

Mast cell density in gastric cancer and its relation to aggressive behavior | 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 Mast cell density in gastric cancer and its relation to aggressive behavior Elham Nazar, Elaheh Farmani, Aysan Nozheh This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6272291/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 Introduction: Stomach cancer is a serious health problem worldwide, as it is one of the most common cancers and the fourth leading cause of cancer-related death. Mast cells perform functions in the immunity system and are a type of discriminated myeloid cell. In this study, we evaluated the correlation between tryptase-positive mast cell density with prognostic histopathological findings in gastric cancer. Materials and Methods: The study was conducted as a cross-sectional study, using tissue samples from 40 patients who underwent radical gastrectomy at Sina Hospital between 2022 and 2023. After histopathological examination and determination of tumor histopathological characteristics, the samples were subjected to immunohistochemical staining using a monoclonal antibody against mast cell tryptase. Results: In this study, the median density of mast cells in tumor tissue was 8/10 high power fields. There was no significant relationship between mast cell density and the number of lymph nodes involved, as well as tumor type, grade, location, and size. Furthermore, there was no significant relationship between mast cell density and tumor vascular invasion or neural invasion. Conclusion: Mast cells have vital roles in normal immune systems and pathological situations. Mast cell density in tumor tissue might be considered for the prognosis of patients before treatment but the function of mast cells has not been completely explained in gastric cancer and needs confirmation to introduce new target therapy. Stomach cancer mast cell density prognosis Figures Figure 1 Introduction Stomach cancer is the fourth most common cancer in the world and the second cause of cancer death [ 1 – 3 ]. Due to poor awareness, gastric malignancies are often diagnosed at a late stage, and because conventional treatments are ineffective, most patients with surgery have a very low survival rate, about five years, and die [ 4 ]. The mainly of solid cancers are related to chronic inflammation and infection [ 5 ]. The exact molecular mechanism of gastric cancer is unknown, and for this reason, it is considered to be a complex multi-step process due to interactions between environment and genetics. Helicobacter pylori infection, Epstein-Barr virus, and dietary habits are considered to be major environmental factors [ 6 ]. Chronic inflammation is an important feature in cancerous tissue, and mast cells have an important role in this inflammation site [ 7 ]. The relationship between the immune system and the development of cancer has been studied for a long time but is still not fully understood [ 8 ]. Gastric cancerous tissue has a combination of stomach tumoral cells and a variety of immune cells, which may exist in the tumoral mass or enter from other tissues [ 9 ]. Tumor outcomes are determined not only by the intrinsic characteristics of the cancer cells as well as by the interactions among tumoral cells and their microenvironment. Tumor-infiltrating immune cells are of great interest, but mast cells are less considered. Current investigations have emphasized the effect of mast cells on cancer cell properties and invasion, but the possible impact of mast cell infiltration remains controversial [ 10 ]. Human mast cells form a highly heterogeneous cell population with different morphologies, mediators, and surface receptors [ 11 ]. Mast cells are a kind of differentiated myeloid cells and have roles in allergic reactions (especially type 1), and in native and developed immunity response by recognizing and eradicating pathogens as well as releasing active immune mediators [ 12 ]. However, the exact function of mast cells in tumor formation and growth remains controversial: mast cell-derived mediators can perform pro-tumor functions, induce tumor growth and spread, or anti-tumor functions, or limit tumor growth [ 13 ]. Previous studies have determined that mast cell infiltration levels are increased in several kinds of tumor tissues of lymphoma and breast cancer which are related to the probability of mast cell function in the growth of the tumor [ 14 ]. Mast cells constitute a small subpopulation of tumor microenvironment (TME) immune cells in tumoral tissue. Mast cells gather both in the tumoral mass and the adjacent tissues. Mast cells affect TME transformation and progression by interrelating with other components of the TME. However, it remains controversial whether mast cells relate to tumor development or activate an immune response to the tumor [ 15 ]. The act of mast cells in gastric adenocarcinoma has been a part of ​​important attention. Mast cell density may be involved in the expansion and prognosis of gastric cancer. We wanted to evaluate the association of mast cell density with the potential for lymph node metastasis and other histopathological findings in gastric cancer to predict aggressive behavior. Materials and Methods In this cross-sectional study, 40 tissue specimens from patients with gastric adenocarcinoma who underwent radical gastrectomy at Sina Hospital during 2022–2023 were subjected to immunohistochemical (IHC) staining using the monoclonal antibody against mast cell tryptase after histopathological examination and determination of tumor pathological characteristics. The association between IHC expression of mast cell tryptase and demographic and histopathological factors, particularly lymph node metastasis, was statistically analyzed. Tissue sections 4 µm thick were prepared from paraffin blocks on glass slides and autoclaved at 60°C overnight to perform IHC staining. The specimens were then immersed in xylenol three times for 15 minutes each, 100 percent alcohol twice for 10 minutes each, and finally 96 percent alcohol twice for 10 minutes. The sample was then cleaned with distilled water before being sterilized for 20 minutes at 95 ℃ and 2 bar pressure with 1 point 5 ml of EDTA buffer (pH 8). The slices were once more washed twice or three times in distilled water before being left out at room temperature for 20 minutes. Next, a peroxidase solution was used to block the internal peroxidase for 10 minutes. After being in contact with the mast cell tryptase monoclonal antibody (DAKO, USA) for 10 minutes, the samples were then washed with TBS buffer and a secondary antibody was added. The samples were once more washed with TBS buffer after 30 minutes, and the DAB chromatin was then exposed to the samples for 10 minutes. After being dehydrated, the sample is prepared for mounting and microscopic examination. The samples were lastly cleaned under running water, stained with hematoxylin, and placed in bicarbonate for 30 seconds. The IHC expression of mast cell tryptase and mast cell density (average in 10 high power fields (hpf)) were associated with demographic and histopathological factors, particularly lymph node metastasis, and statistical analysis was performed after the stained slides were examined by two pathologists. In tumor tissue, mast cell tryptase staining in cytoplasmic infiltrating immune cells is considered positive (Fig. 1 ). Also, the study has been reported in line with the STROCSS 2021 criteria [ 15 ]. Statistical analysis A P value of 0.05 or less was regarded as significant for all analyses, which were conducted using the statistical package for social sciences software (SPSS version 26; IBM Company). Baseline data were checked for normality using the Kolmogorov-Smirnov test and probability plots. Continuous data were shown as mean ± SD, whereas categorical variables were reported in percent (%). Levene's test for equal variance and Fisher's exact test were used, respectively, to compare continuous and categorical variables using an independent t-test. Results According to the study, there were 40 patients with gastric cancer with an average age of 63 years (minimum age 39 years and maximum age 84 years), of which 75% were men and 25% were women. In all patients, 80% of the tumors were intestinal type and 20% were signet ring. The lymphovascular invasion occurred in 50% of tumors. Additionally, 58% of tumors showed no neural invasion and 42% showed neural invasion. Patients had an average of 5 lymph nodes that were affected (the range was 0 to 32). In addition, the tumor tissue had an average mast cell density of 8/10 hpfs (the minimum and maximum values were 2/10 hpfs and 30/10 hpfs, respectively). Tumor sizes ranged from 1 to 11 centimeters. Baseline characteristic data was summarized in Table 1 . This study discovered no statistically significant relationship between mast cell density and lymphovascular invasion (p = 0.299), the number of involved lymph nodes (p = 0.454), or tumor stage (p = 0.366). Further research revealed no connection between mast cell density and neural invasion (p = 0.227), tumor size (p = 0.455), or tumor type (p = 0.55). Table 1 Baseline characteristics of study subjects. Mean age, year 62.7 ± 10.80 Gender, % Male 30 (75.0) Female 10 (25.0) Histologic subtype intestinal 32 (80.0) Signet-ring 8 (20.0) Histologic grade I 12 (30.0) II 11 (27.5) III 17 (42.5) Tumor location Antrum 24 (60.0) Body 3 (7.5) Cardia 11 (27.5) Fundus 2 (5.0) Neural invasion, % 17 (42.5) Lymphovascular invasion, % 20 (50.0) Tumor stage, % I 3 (7.5) II 7 (17.5) III 23 (57.5) IV 7 (17.5) Mean size, cm 4.62 ± 2.78 Total lymph nodes dissected (mean ± SD) 17.7 ± 9.5 Involved lymph nodes dissected(mean ± SD) 4.9 ± 6.7 Mean mast cell tryptase infiltrating cells 8/ 10hpfs Discussion As a solid tumor, gastric cancer includes immune cells as well as stromal, epithelial, and endothelial cells in addition to tumor cells. Infiltrating immune cells and stromal cells are two key non-tumor cells in the TME of gastric cancer. Numerous studies have demonstrated their importance for understanding the onset and progression of gastric cancer as well as its prognosis and drug resistance [ 16 ]. Mast cells are prominently found in solid tumors and show unique phenotypes depending on the tumor microenvironment. Their exact mode of communication in gastric cancer is still largely unknown. In the majority of tumors, such as thyroid [ 17 ], gastric [ 18 , 19 ], pancreas [ 20 – 22 ], bladder [ 23 ], and colorectal [ 24 , 25 ] cancers, hepatocellular carcinoma [ 26 , 27 ], Merkel cell carcinoma [ 28 ], Hodgkin’s [ 29 ] and non-Hodgkin’s lymphoma [ 30 ] and plasmacytoma [ 31 ], mast cells conferred poor prognosis. Mast cells might act as an antitumorigenic factor in breast cancer [ 32 ]. These results suggest that mast cells' involvement in cancer depends on the tumor. Mast cells in gastric cancer have only been the subject of a small number of studies. We still don't know how mast cells affect the onset and progression of gastric cancer or how to control inflammation. LV Y, et al. discovered that patients with gastric cancer had tumors with noticeably more mast cells. Mast cell levels increased as the tumor developed and independently predicted a lower overall survival rate [ 33 ]. A recent study by Sammarco G, et al. showed gastric cancer patients have higher mast cell densities, which are connected with angiogenesis, the number of lymph nodes that have spread the disease, and the survival of these patients. By releasing angiogenic and lymphangiogenic factors, mast cells play a protumorigenic role in gastric cancer [ 34 ]. In another study by Ribatti D et al., commercially available samples from 30 patients undergoing curative gastrectomy were examined immunohistochemically using an anti-CD31 antibody to stain endothelial cells and an anti-tryptase and anti-chymase antibody to stain the mast cells. The results showed that stage IV gastric cancer has a higher degree of vascularity than other stages and tryptase and chymase-positive mast cells increase concomitantly with the degree of malignancy, even though the density of chymase-positive mast cells was significantly lower than those of tryptase-mast cell positive and is highly correlated with the degree of angiogenesis. This study demonstrated that angiogenesis and progression of gastric cancer patients are related to mast cell density. Understanding the mechanisms of gastric cancer angiogenesis provides a basis for a biological approach to the development of anti-angiogenic therapy in patients with this malignancy [ 35 ]. Zhong B et al. study showed there were more mast cells in the gastric cancer tissue than in the surrounding cancer tissue. Mast cell enrichment in gastric cancer tissue was found by flow cytometry and IHC staining, which may have a role in the growth of the disease [ 36 ]. The significant anti-tumor activity of mast cells revealed an as-yet unreported effect on upregulating tumor cell suppressor genes and downregulating pro-tumor genes [ 37 ]. Liu YL et al. study showed mast cell density was inversely related to the number of lymph node metastases and the depth of tumor invasion in patients with stomach cancer [ 38 ]. To better understand the functions of mast cells in tumoral tissue, preclinical evaluations were used in animal models with dysfunctional c-KIT. These studies also suggest that mast cells may facilitate vascularization, metastasis, and release of growth factors and enzymes from the tumor. These results thus show the possibility of new therapeutic approaches via the targeting of mast cells for precise chemotherapy or combined therapies. Animal models will make it easier to determine whether mast cells are harmful or helpful in the progression of cancer, though mouse data will always need to be properly validated in clinical patient settings [ 39 ]. In various pre-clinical models of solid tumors, mast cell-stabilizing agents like cromolyn sodium have been applied. Treatment with cromolyn slowed tumor growth in a mouse xenograft model of thyroid cancer [ 40 ]. In the peritoneal dissemination of gastric cancer, Interleukin-17 (IL-17) produced by mast cells aids in tumor fibrosis. A potential therapeutic approach to manage organ fibrosis involves preventing mast cell degranulation [ 41 ]. It appears that mast cell density is diverse in tumors that vary from all societies. In our study, the activity of these immune cells in predicting tumor histopathological and behavioral outcomes and consequent adverse cancer outcomes was completely questioned. In general, it can be assumed that in our culture the value of mast cell density in the assessment and estimation of aggressive behavior in gastric cancer is low, but it should be noted that the small sample size of the study could potentially limit the reliability of the results. Thus, further evaluation with a larger sample is strongly recommended. Conclusion We need to be more accurate than tumor characteristics in predicting the prognosis of gastric cancer patients. Mast cells are frequently found in a tumor's microenvironment, which suggests that they play a role in the development of cancer from chronic inflammation. Mast cell involvement in the solid malignancy microenvironment is still debatable. We also look forward to mast cells' potential role in cancer immunotherapy, which could build on the success of current cancer therapies. Our data need confirmation in a larger patient cohort. Abbreviations Tumor microenvironment TME Immunohistochemical IHC High power fields hpf Declarations Ethics approval and consent to participate: All individuals who contributed samples to this study voluntarily provided their informed consent to participate. They were thoroughly briefed on the research’s objectives and potential advantages. Participants were guaranteed the right to withdraw at any point without facing any repercussions. Written consent was secured from all participants before their inclusion in the study. This study was conducted in accordance with the ethical principles outlined in the Declaration of Helsinki. Ethical approval was obtained from the ethics committee of Tehran University of Medical Sciences, Sina Hospital. Consent for publication: Not Applicable Availability of data and materials: The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. Competing interests: We declare that we have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. also, we declare that we are preparing articles in our personal capacity. Funding: No source is to be stated. Authors' contributions: Elham Nazar: Conceptualization, Methodology, Project administration, Writing - original draft, Supervision Elaheh Farmani: Investigation, Validation, Writing - review & editing, Resources Aysan Nozheh (Corresponding author): Writing - review & editing, Writing - original draft, Data curation, Software, Formal analysis Acknowledgements: Not applicable References Jemal A et al. Global cancer statistics. CA: A Cancer Journal for Clinicians, 2011. 61(2): pp. 69–90. Lott PC, Carvajal-Carmona LG. Resolving gastric cancer etiology: an update in genetic predisposition. Lancet Gastroenterol Hepatol. 2018;3(12):874–83. Van Cutsem E, et al. Gastric cancer. 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Few, but efficient: the role of mast cells in breast cancer and other solid tumors. Cancer Res. 2022;82(8):1439–47. Varricchi G, et al. Human mast cells and basophils- How are they similar how are they different? Immunol Rev. 2018;282(1):8–34. Theoharides TC, et al. Mast cells and inflammation. Biochim Biophys Acta. 2012;1822(1):21–33. Molfetta R, Paolini R. The controversial role of intestinal mast cells in colon cancer. Cells. 2023;12(3):459. Raica M, et al. Interplay between mast cells and lymphatic vessels in different molecular types of breast cancer. Anticancer Res. 2013;33(3):957–63. Shi S et al. Focus on mast cells in the tumor microenvironment: Current knowledge and future directions. Biochimica et Biophysica Acta (BBA)-Reviews on Cancer, 2022: p. 188845. Cao W, et al. A novel signature based on CeRNA and immune status predicts prognostic risk and drug sensitivity in gastric cancer patients. Front Immunol. 2022;13:951135. Visciano C, et al. 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Clin Cancer Res. 2011;17(22):7015–23. Rao Q, et al. Recruited mast cells in the tumor microenvironment enhance bladder cancer metastasis via modulation of ERβ/CCL2/CCR2 EMT/MMP9 signals. Oncotarget. 2016;7(7):7842–55. Ammendola M, et al. Correlation between serum tryptase, mast cells positive to tryptase and microvascular density in colorectal cancer patients: possible biological-clinical significance. PLoS ONE. 2014;9(6):e99512. Malfettone A, et al. High density of tryptase-positive mast cells in human colorectal cancer: a poor prognostic factor related to protease-activated receptor 2 expression. J Cell Mol Med. 2013;17(8):1025–37. Tu JF, et al. Mast Cells Comprise the Major of Interleukin 17-Producing Cells and Predict a Poor Prognosis in Hepatocellular Carcinoma. Med (Baltim). 2016;95(13):e3220. Ammendola M, et al. Mast cells positive to tryptase, endothelial cells positive to protease-activated receptor-2, and microvascular density correlate among themselves in hepatocellular carcinoma patients who have undergone surgery. Onco Targets Ther. 2016;9:4465–71. Beer TW, Ng LB, Murray K. Mast cells have prognostic value in Merkel cell carcinoma. Am J Dermatopathol. 2008;30(1):27–30. Andersen MD, et al. Tumour-associated mast cells in classical Hodgkin's lymphoma: correlation with histological subtype, other tumor-infiltrating inflammatory cell subsets, and outcome. Eur J Haematol. 2016;96(3):252–9. Franco G, et al. Bone marrow stroma CD40 expression correlates with inflammatory mast cell infiltration and disease progression in splenic marginal zone lymphoma. Blood. 2014;123(12):1836–49. Nakayama T, Yao L, Tosato G. Mast cell-derived angiopoietin-1 plays a critical role in the growth of plasma cell tumors. J Clin Invest. 2004;114(9):1317–25. Amini R-M, et al. Mast cells and eosinophils in invasive breast carcinoma. BMC Cancer. 2007;7(1):165. Lv Y-p, et al. Degranulation of mast cells induced by gastric cancer-derived adrenomedullin prompts gastric cancer progression. Volume 9. Cell death & disease; 2018. p. 1034. 10. Sammarco G et al. Mast Cells, Angiogenesis and Lymphangiogenesis in Human Gastric Cancer. Int J Mol Sci, 2019. 20(9). Ribatti D, et al. Mast cells and angiogenesis in gastric carcinoma. Int J Exp Pathol. 2010;91(4):350–6. Zhong B, et al. Association of mast cell infiltration with gastric cancer progression. Oncol Lett. 2018;15(1):755–64. Fereydouni M, et al. Human tumor-targeted cytotoxic mast cells for cancer immunotherapy. Front Oncol. 2022;12:871390. Liu YL, et al. [Effects of mast cell infiltration on the development and metastasis of gastric carcinoma]. Di Yi Jun Yi Da Xue Xue Bao. 2005;25(7):809–11. Sobiepanek A, et al. The multifaceted roles of mast cells in immune homeostasis, infections and cancers. Int J Mol Sci. 2022;23(4):2249. Solimando AG, Desantis V, Ribatti D. Mast cells and interleukins. Int J Mol Sci. 2022;23(22):14004. Gunjigake K, et al. Interleukin-17A derived from mast cells contributes to fibrosis in gastric cancer with peritoneal dissemination. Gastric Cancer. 2021;24:31–44. Additional Declarations No competing interests reported. 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-6272291","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":448150437,"identity":"ffd7fc6d-06e1-4993-aeb6-263008278fcf","order_by":0,"name":"Elham Nazar","email":"","orcid":"","institution":"Tehran University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Elham","middleName":"","lastName":"Nazar","suffix":""},{"id":448150439,"identity":"b74c19b4-7086-4a2d-b62c-8ffbd5afb8c8","order_by":1,"name":"Elaheh Farmani","email":"","orcid":"","institution":"Tehran University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Elaheh","middleName":"","lastName":"Farmani","suffix":""},{"id":448150442,"identity":"3d7b85a5-1edc-4f73-b3ed-0b9ecf2691bd","order_by":2,"name":"Aysan Nozheh","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA+ElEQVRIiWNgGAWjYLACxgaGBH4QIwEhJkFYi2QDWIsBCVoMDoCZBvhVggB/A+/Bhz932OQZ3z78+MMDhj+Ja/sPMH74wWCRj0uLxAG+ZAPJM2nFZufSDAyADkvcdiOBWbKHQcKyAZee+2/MJAzbDiduO8NgkADRwsAgDTQLpxPlD/CYSSS2/U/c3MP+4QBYy/kDzL/xaTEAaTnYdiBxAw+PYQNYy4EENry2GB7gMTZsbEtOnHGGpxgYbsbG224ktln2GODWIneAx/Dhzza7xP4e9s0ff1TIyW47f/jwjR8VdUQEN8SdIAIYTcTEzygYBaNgFIwC3AAAn5NTRELpX3sAAAAASUVORK5CYII=","orcid":"","institution":"Tehran University of Medical Sciences","correspondingAuthor":true,"prefix":"","firstName":"Aysan","middleName":"","lastName":"Nozheh","suffix":""}],"badges":[],"createdAt":"2025-03-20 20:23:07","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6272291/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6272291/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":82123925,"identity":"740a069a-532a-4a11-ba6f-53e88fa75fbd","added_by":"auto","created_at":"2025-05-07 03:36:34","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1950364,"visible":true,"origin":"","legend":"\u003cp\u003eHistopathological examination shows gastric adenocarcinoma by H\u0026amp;E staining (left) and immunohistochemical staining of mast cell tryptase in infiltrating cells in the tumor (right) at magnification x400.\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-6272291/v1/4f6c7b23fdf646dc08e0ac01.jpeg"},{"id":98380736,"identity":"6ac6a005-af1c-4b1d-9fae-5a3c99cda84d","added_by":"auto","created_at":"2025-12-17 07:40:48","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2371661,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6272291/v1/71f3465a-bd1f-4a68-acaa-8bd4e069ae74.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Mast cell density in gastric cancer and its relation to aggressive behavior","fulltext":[{"header":"Introduction","content":"\u003cp\u003eStomach cancer is the fourth most common cancer in the world and the second cause of cancer death [\u003cspan additionalcitationids=\"CR2\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Due to poor awareness, gastric malignancies are often diagnosed at a late stage, and because conventional treatments are ineffective, most patients with surgery have a very low survival rate, about five years, and die [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. The mainly of solid cancers are related to chronic inflammation and infection [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. The exact molecular mechanism of gastric cancer is unknown, and for this reason, it is considered to be a complex multi-step process due to interactions between environment and genetics. Helicobacter pylori infection, Epstein-Barr virus, and dietary habits are considered to be major environmental factors [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Chronic inflammation is an important feature in cancerous tissue, and mast cells have an important role in this inflammation site [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. The relationship between the immune system and the development of cancer has been studied for a long time but is still not fully understood [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Gastric cancerous tissue has a combination of stomach tumoral cells and a variety of immune cells, which may exist in the tumoral mass or enter from other tissues [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Tumor outcomes are determined not only by the intrinsic characteristics of the cancer cells as well as by the interactions among tumoral cells and their microenvironment. Tumor-infiltrating immune cells are of great interest, but mast cells are less considered. Current investigations have emphasized the effect of mast cells on cancer cell properties and invasion, but the possible impact of mast cell infiltration remains controversial [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Human mast cells form a highly heterogeneous cell population with different morphologies, mediators, and surface receptors [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Mast cells are a kind of differentiated myeloid cells and have roles in allergic reactions (especially type 1), and in native and developed immunity response by recognizing and eradicating pathogens as well as releasing active immune mediators [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. However, the exact function of mast cells in tumor formation and growth remains controversial: mast cell-derived mediators can perform pro-tumor functions, induce tumor growth and spread, or anti-tumor functions, or limit tumor growth [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Previous studies have determined that mast cell infiltration levels are increased in several kinds of tumor tissues of lymphoma and breast cancer which are related to the probability of mast cell function in the growth of the tumor [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Mast cells constitute a small subpopulation of tumor microenvironment (TME) immune cells in tumoral tissue. Mast cells gather both in the tumoral mass and the adjacent tissues. Mast cells affect TME transformation and progression by interrelating with other components of the TME. However, it remains controversial whether mast cells relate to tumor development or activate an immune response to the tumor [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. The act of mast cells in gastric adenocarcinoma has been a part of ​​important attention. Mast cell density may be involved in the expansion and prognosis of gastric cancer. We wanted to evaluate the association of mast cell density with the potential for lymph node metastasis and other histopathological findings in gastric cancer to predict aggressive behavior.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003eIn this cross-sectional study, 40 tissue specimens from patients with gastric adenocarcinoma who underwent radical gastrectomy at Sina Hospital during 2022\u0026ndash;2023 were subjected to immunohistochemical (IHC) staining using the monoclonal antibody against mast cell tryptase after histopathological examination and determination of tumor pathological characteristics. The association between IHC expression of mast cell tryptase and demographic and histopathological factors, particularly lymph node metastasis, was statistically analyzed.\u003c/p\u003e \u003cp\u003eTissue sections 4 \u0026micro;m thick were prepared from paraffin blocks on glass slides and autoclaved at 60\u0026deg;C overnight to perform IHC staining. The specimens were then immersed in xylenol three times for 15 minutes each, 100 percent alcohol twice for 10 minutes each, and finally 96 percent alcohol twice for 10 minutes. The sample was then cleaned with distilled water before being sterilized for 20 minutes at 95 ℃ and 2 bar pressure with 1 point 5 ml of EDTA buffer (pH 8). The slices were once more washed twice or three times in distilled water before being left out at room temperature for 20 minutes. Next, a peroxidase solution was used to block the internal peroxidase for 10 minutes. After being in contact with the mast cell tryptase monoclonal antibody (DAKO, USA) for 10 minutes, the samples were then washed with TBS buffer and a secondary antibody was added. The samples were once more washed with TBS buffer after 30 minutes, and the DAB chromatin was then exposed to the samples for 10 minutes. After being dehydrated, the sample is prepared for mounting and microscopic examination. The samples were lastly cleaned under running water, stained with hematoxylin, and placed in bicarbonate for 30 seconds. The IHC expression of mast cell tryptase and mast cell density (average in 10 high power fields (hpf)) were associated with demographic and histopathological factors, particularly lymph node metastasis, and statistical analysis was performed after the stained slides were examined by two pathologists. In tumor tissue, mast cell tryptase staining in cytoplasmic infiltrating immune cells is considered positive (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Also, the study has been reported in line with the STROCSS 2021 criteria [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eA P value of 0.05 or less was regarded as significant for all analyses, which were conducted using the statistical package for social sciences software (SPSS version 26; IBM Company). Baseline data were checked for normality using the Kolmogorov-Smirnov test and probability plots. Continuous data were shown as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD, whereas categorical variables were reported in percent (%). Levene's test for equal variance and Fisher's exact test were used, respectively, to compare continuous and categorical variables using an independent t-test.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eAccording to the study, there were 40 patients with gastric cancer with an average age of 63 years (minimum age 39 years and maximum age 84 years), of which 75% were men and 25% were women. In all patients, 80% of the tumors were intestinal type and 20% were signet ring. The lymphovascular invasion occurred in 50% of tumors. Additionally, 58% of tumors showed no neural invasion and 42% showed neural invasion. Patients had an average of 5 lymph nodes that were affected (the range was 0 to 32). In addition, the tumor tissue had an average mast cell density of 8/10 hpfs (the minimum and maximum values were 2/10 hpfs and 30/10 hpfs, respectively). Tumor sizes ranged from 1 to 11 centimeters. Baseline characteristic data was summarized in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. This study discovered no statistically significant relationship between mast cell density and lymphovascular invasion (p\u0026thinsp;=\u0026thinsp;0.299), the number of involved lymph nodes (p\u0026thinsp;=\u0026thinsp;0.454), or tumor stage (p\u0026thinsp;=\u0026thinsp;0.366). Further research revealed no connection between mast cell density and neural invasion (p\u0026thinsp;=\u0026thinsp;0.227), tumor size (p\u0026thinsp;=\u0026thinsp;0.455), or tumor type (p\u0026thinsp;=\u0026thinsp;0.55).\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\u003eBaseline characteristics of study subjects.\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=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMean age, year\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.7\u0026thinsp;\u0026plusmn;\u0026thinsp;10.80\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGender, %\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=\"left\" colname=\"c2\"\u003e \u003cp\u003e30 (75.0)\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\u003e10 (25.0)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHistologic subtype\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\u003eintestinal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e32 (80.0)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSignet-ring\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8 (20.0)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHistologic grade\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\u003eI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e12 (30.0)\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 (27.5)\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\u003e17 (42.5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTumor location\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\u003eAntrum\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e24 (60.0)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBody\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3 (7.5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCardia\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11 (27.5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFundus\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2 (5.0)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNeural invasion, %\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e17 (42.5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLymphovascular invasion, %\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e20 (50.0)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTumor stage, %\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\u003eI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3 (7.5)\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\u003e7 (17.5)\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\u003e23 (57.5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIV\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7 (17.5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMean size, cm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.62\u0026thinsp;\u0026plusmn;\u0026thinsp;2.78\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal lymph nodes dissected (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e17.7\u0026thinsp;\u0026plusmn;\u0026thinsp;9.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eInvolved lymph nodes dissected(mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.9\u0026thinsp;\u0026plusmn;\u0026thinsp;6.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMean mast cell tryptase infiltrating cells\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\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8/ 10hpfs\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eAs a solid tumor, gastric cancer includes immune cells as well as stromal, epithelial, and endothelial cells in addition to tumor cells. Infiltrating immune cells and stromal cells are two key non-tumor cells in the TME of gastric cancer. Numerous studies have demonstrated their importance for understanding the onset and progression of gastric cancer as well as its prognosis and drug resistance [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Mast cells are prominently found in solid tumors and show unique phenotypes depending on the tumor microenvironment. Their exact mode of communication in gastric cancer is still largely unknown. In the majority of tumors, such as thyroid [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e], gastric [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e], pancreas [\u003cspan additionalcitationids=\"CR21\" citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e], bladder [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e], and colorectal [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e] cancers, hepatocellular carcinoma [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e], Merkel cell carcinoma [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e], Hodgkin\u0026rsquo;s [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e] and non-Hodgkin\u0026rsquo;s lymphoma [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e] and plasmacytoma [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e], mast cells conferred poor prognosis. Mast cells might act as an antitumorigenic factor in breast cancer [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. These results suggest that mast cells' involvement in cancer depends on the tumor. Mast cells in gastric cancer have only been the subject of a small number of studies. We still don't know how mast cells affect the onset and progression of gastric cancer or how to control inflammation.\u003c/p\u003e \u003cp\u003eLV Y, et al. discovered that patients with gastric cancer had tumors with noticeably more mast cells. Mast cell levels increased as the tumor developed and independently predicted a lower overall survival rate [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. A recent study by Sammarco G, et al. showed gastric cancer patients have higher mast cell densities, which are connected with angiogenesis, the number of lymph nodes that have spread the disease, and the survival of these patients. By releasing angiogenic and lymphangiogenic factors, mast cells play a protumorigenic role in gastric cancer [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. In another study by Ribatti D et al., commercially available samples from 30 patients undergoing curative gastrectomy were examined immunohistochemically using an anti-CD31 antibody to stain endothelial cells and an anti-tryptase and anti-chymase antibody to stain the mast cells. The results showed that stage IV gastric cancer has a higher degree of vascularity than other stages and tryptase and chymase-positive mast cells increase concomitantly with the degree of malignancy, even though the density of chymase-positive mast cells was significantly lower than those of tryptase-mast cell positive and is highly correlated with the degree of angiogenesis. This study demonstrated that angiogenesis and progression of gastric cancer patients are related to mast cell density. Understanding the mechanisms of gastric cancer angiogenesis provides a basis for a biological approach to the development of anti-angiogenic therapy in patients with this malignancy [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Zhong B et al. study showed there were more mast cells in the gastric cancer tissue than in the surrounding cancer tissue. Mast cell enrichment in gastric cancer tissue was found by flow cytometry and IHC staining, which may have a role in the growth of the disease [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. The significant anti-tumor activity of mast cells revealed an as-yet unreported effect on upregulating tumor cell suppressor genes and downregulating pro-tumor genes [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. Liu YL et al. study showed mast cell density was inversely related to the number of lymph node metastases and the depth of tumor invasion in patients with stomach cancer [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. To better understand the functions of mast cells in tumoral tissue, preclinical evaluations were used in animal models with dysfunctional c-KIT. These studies also suggest that mast cells may facilitate vascularization, metastasis, and release of growth factors and enzymes from the tumor. These results thus show the possibility of new therapeutic approaches via the targeting of mast cells for precise chemotherapy or combined therapies. Animal models will make it easier to determine whether mast cells are harmful or helpful in the progression of cancer, though mouse data will always need to be properly validated in clinical patient settings [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. In various pre-clinical models of solid tumors, mast cell-stabilizing agents like cromolyn sodium have been applied. Treatment with cromolyn slowed tumor growth in a mouse xenograft model of thyroid cancer [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. In the peritoneal dissemination of gastric cancer, Interleukin-17 (IL-17) produced by mast cells aids in tumor fibrosis. A potential therapeutic approach to manage organ fibrosis involves preventing mast cell degranulation [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. It appears that mast cell density is diverse in tumors that vary from all societies. In our study, the activity of these immune cells in predicting tumor histopathological and behavioral outcomes and consequent adverse cancer outcomes was completely questioned. In general, it can be assumed that in our culture the value of mast cell density in the assessment and estimation of aggressive behavior in gastric cancer is low, but it should be noted that the small sample size of the study could potentially limit the reliability of the results. Thus, further evaluation with a larger sample is strongly recommended.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eWe need to be more accurate than tumor characteristics in predicting the prognosis of gastric cancer patients. Mast cells are frequently found in a tumor's microenvironment, which suggests that they play a role in the development of cancer from chronic inflammation. Mast cell involvement in the solid malignancy microenvironment is still debatable. We also look forward to mast cells' potential role in cancer immunotherapy, which could build on the success of current cancer therapies. Our data need confirmation in a larger patient cohort.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eTumor microenvironment\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eTME\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eImmunohistochemical\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eIHC\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eHigh power fields\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ehpf\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll individuals who contributed samples to this study voluntarily provided their informed consent to participate. They were thoroughly briefed on the research\u0026rsquo;s objectives and potential advantages. Participants were guaranteed the right to withdraw at any point without facing any repercussions. Written consent was secured from all participants before their inclusion in the study. This study was conducted in accordance with the ethical principles outlined in the Declaration of Helsinki. Ethical approval was obtained from the ethics committee of Tehran University of Medical Sciences, Sina Hospital.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot Applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests:\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eWe declare that we have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. also, we declare that we are preparing articles in our personal capacity.\u003cbr\u003e\u0026nbsp;\u003cstrong\u003eFunding:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo source is to be stated.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eElham Nazar: Conceptualization, Methodology, Project administration, Writing - original draft, Supervision\u003c/p\u003e\n\u003cp\u003eElaheh Farmani: Investigation, Validation, Writing - review \u0026amp; editing, Resources\u003c/p\u003e\n\u003cp\u003eAysan Nozheh (Corresponding author): Writing - review \u0026amp; editing, Writing - original draft, Data curation, Software, Formal analysis\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eJemal A et al. \u003cem\u003eGlobal cancer statistics.\u003c/em\u003e CA: A Cancer Journal for Clinicians, 2011. 61(2): pp. 69\u0026ndash;90.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLott PC, Carvajal-Carmona LG. 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[Effects of mast cell infiltration on the development and metastasis of gastric carcinoma]. Di Yi Jun Yi Da Xue Xue Bao. 2005;25(7):809\u0026ndash;11.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSobiepanek A, et al. The multifaceted roles of mast cells in immune homeostasis, infections and cancers. Int J Mol Sci. 2022;23(4):2249.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSolimando AG, Desantis V, Ribatti D. Mast cells and interleukins. Int J Mol Sci. 2022;23(22):14004.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGunjigake K, et al. Interleukin-17A derived from mast cells contributes to fibrosis in gastric cancer with peritoneal dissemination. Gastric Cancer. 2021;24:31\u0026ndash;44.\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":"Stomach cancer, mast cell density, prognosis","lastPublishedDoi":"10.21203/rs.3.rs-6272291/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6272291/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIntroduction: Stomach cancer is a serious health problem worldwide, as it is one of the most common cancers and the fourth leading cause of cancer-related death. Mast cells perform functions in the immunity system and are a type of discriminated myeloid cell. In this study, we evaluated the correlation between tryptase-positive mast cell density with prognostic histopathological findings in gastric cancer.\u003c/p\u003e \u003cp\u003eMaterials and Methods: The study was conducted as a cross-sectional study, using tissue samples from 40 patients who underwent radical gastrectomy at Sina Hospital between 2022 and 2023. After histopathological examination and determination of tumor histopathological characteristics, the samples were subjected to immunohistochemical staining using a monoclonal antibody against mast cell tryptase.\u003c/p\u003e \u003cp\u003eResults: In this study, the median density of mast cells in tumor tissue was 8/10 high power fields. There was no significant relationship between mast cell density and the number of lymph nodes involved, as well as tumor type, grade, location, and size. Furthermore, there was no significant relationship between mast cell density and tumor vascular invasion or neural invasion.\u003c/p\u003e \u003cp\u003eConclusion: Mast cells have vital roles in normal immune systems and pathological situations. Mast cell density in tumor tissue might be considered for the prognosis of patients before treatment but the function of mast cells has not been completely explained in gastric cancer and needs confirmation to introduce new target therapy.\u003c/p\u003e","manuscriptTitle":"Mast cell density in gastric cancer and its relation to aggressive behavior","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-07 03:36:30","doi":"10.21203/rs.3.rs-6272291/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":"4bc52f08-6024-4a0d-be53-58226d734c66","owner":[],"postedDate":"May 7th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-12-17T07:40:22+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-07 03:36:30","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6272291","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6272291","identity":"rs-6272291","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}