ISLR interacts with MGAT5 to promote the malignant progression of human gastric cancer AGS cells

Document Type : Original Article

Authors

Department of Gastroenterology Surgery, Yichang Central People’s Hospital, The First College of Clinical Medical Science, China Three Gorges University, Yichang 443000, Hubei, China

Abstract

Objective(s): Gastric cancer is a common malignant tumor with high morbidity and mortality. The present study aimed to investigate the role of the immunoglobulin superfamily containing leucine-rich repeat (ISLR) gene in gastric cancer and examine whether ISLR could interact with N-acetylglucosaminyltransferase V (MGAT5) to affect the malignant progression of gastric cancer. 
Materials and Methods: The expression of ISLR and MGAT5 in human normal gastric epithelial cells and human gastric cancer cells, and the transfection efficiency of ISLR interference plasmids and MGAT5 overexpression plasmids were all detected by reverse transcription-quantitative PCR (RT-qPCR) and western blot. The viability, proliferation, migration and invasion, and epithelial-mesenchymal transition (EMT) of gastric cancer cells after indicated transfection were detected by Cell counting kit-8 (CCK-8) assay, 5-ethynyl-2’-deoxyuridine (EdU) staining, wound healing assay, and transwell assay. The interaction between ISLR and MGAT5 was confirmed by co-immunoprecipitation. The expression of proteins related to migration, invasion, and EMT was detected by immunofluorescence and western blot.
Results: As a result, ISLR was highly expressed in gastric cancer and was associated with poor prognosis. Interference with ISLR inhibited the viability, proliferation, migration, invasion, and EMT of gastric cancer cells. ISLR interacted with MGAT5 in gastric cancer cells. MGAT5 overexpression weakened the effects of ISLR knockdown on suppressing the viability, proliferation, migration, invasion, and EMT of gastric cancer cells.
Conclusion: ISLR interacted with MGAT5 to promote the malignant progression of gastric cancer.

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Main Subjects


1. Johnston FM, Beckman M. Updates on management of gastric cancer. Curr Oncol Rep 2019; 21:6775.
2. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2021; 71:209-249.
3. Smyth EC, Nilsson M, Grabsch HI, van Grieken NC, Lordick F. Gastric cancer. Lancet 2020; 396:635-648.
4. Lind L, Sandström H, Wahlin A, Eriksson M, Nilsson-Sojka B, Sikström C, et al. Localization of the gene for congenital dyserythropoietic anemia type III, CDAN3, to chromosome 15q21-q25. Hum Mol Genet 1995; 4:109-112.
5. Li S, Zhao W, Sun M. An analysis regarding the association between the ISLR gene and gastric carcinogenesis. Front Genet 2020; 11:620-633.
6. Zhang P, Li Z, Yang G. Silencing of ISLR inhibits tumour progression and glycolysis by inactivating the IL‑6/JAK/STAT3 pathway in non‑small cell lung cancer. Int J Mol Med 2021; 48:222-234.
7. Chi C, Liu T, Yang S, Wang B, Han W, Li J. ISLR affects colon cancer progression by regulating the epithelial-mesenchymal transition signaling pathway. Anticancer Drugs 2022; 33:e670-e679.
8. Xu J, Tang Y, Sheng X, Tian Y, Deng M, Du S, et al. Secreted stromal protein ISLR promotes intestinal regeneration by suppressing epithelial Hippo signaling. Embo J 2020; 39:e103255-e103273.
9. Tian S, Peng P, Li J, Deng H, Zhan N, Zeng Z, et al. SERPINH1 regulates EMT and gastric cancer metastasis via the Wnt/β-catenin signaling pathway. Aging 2020; 12:3574-3593.
10. Lukaszewicz-Zając M, Mroczko B, Szmitkowski M. Gastric cancer-The role of matrix metalloproteinases in tumor progression. Clin Chim Acta 2011; 412:1725-1730.
11. Peng Z, Wang CX, Fang EH, Wang GB, Tong Q. Role of epithelial-mesenchymal transition in gastric cancer initiation and progression. World J Gastroenterol 2014; 20:5403-5410.
12. Huang L, Wu RL, Xu AM. Epithelial-mesenchymal transition in gastric cancer. Am J Transl Res 2015; 7:2141-2158.
13. Saito T, Miyoshi E, Sasai K, Nakano N, Eguchi H, Honke K, et al. A secreted type of beta 1,6-N-acetylglucosaminyltransferase V (GnT-V) induces tumor angiogenesis without mediation of glycosylation: A novel function of GnT-V distinct from the original glycosyltransferase activity. J Biol Chem 2002; 277:17002-17008.
14. Taniguchi N, Ihara S, Saito T, Miyoshi E, Ikeda Y, Honke K. Implication of GnT-V in cancer metastasis: a glycomic approach for identification of a target protein and its unique function as an angiogenic cofactor. Glycoconj J 2001; 18:859-865.
15. Wang R, Fan Q, Zhang J, Zhang X, Kang Y, Wang Z. Hydrogen sulfide demonstrates promising antitumor efficacy in gastric carcinoma by targeting MGAT5. Transl Oncol 2018; 11:900-910.
16. Hassani Z, Saleh A, Turpault S, Khiati S, Morelle W, Vignon J, et al. Phostine PST3.1a Targets MGAT5 and inhibits glioblastoma-initiating cell invasiveness and proliferation. Mol Cancer Res 2017; 15:1376-1387.
17. Yang Y, Wu J, Liu F, He J, Wu F, Chen J, et al. IGF2BP1 promotes the liver cancer stem cell phenotype by regulating MGAT5 mRNA stability by m6A RNA methylation. Stem Cells Dev 2021; 30:1115-1125.
18. Yan G, Li Y, Zhan L, Sun S, Yuan J, Wang T, et al. Decreased miR-124-3p promoted breast cancer proliferation and metastasis by targeting MGAT5. Am J Cancer Res 2019; 9:585-596.
19. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25:402-408.
20. Kim JH, Kim SS, Lee JH, Jung DH, Cheung DY, Chung WC, et al. Early detection is important to reduce the economic burden of gastric cancer. J Gastric Cancer 2018; 18:82-89.
21. Kim HJ, Oh SC. Novel systemic therapies for advanced gastric cancer. J Gastric Cancer 2018; 18:1-19.
22. Ichikawa H, Nagahashi M, Shimada Y, Hanyu T, Ishikawa T, Kameyama H, et al. Actionable gene-based classification toward precision medicine in gastric cancer. Genome Med 2017; 9:93-104.
23. Tsukamoto Y, Uchida T, Karnan S, Noguchi T, Nguyen LT, Tanigawa M, et al. Genome-wide analysis of DNA copy number alterations and gene expression in gastric cancer. J Pathol 2008; 216:471-482.
24. Deng X, Xiao Q, Liu F, Zheng C. A gene expression-based risk model reveals prognosis of gastric cancer. PeerJ 2018; 6:e4204-215.
25. Zheng HC, Li J, Shen DF, Yang XF, Zhao S, Wu YZ, et al. BTG1 expression correlates with pathogenesis, aggressive behaviors and prognosis of gastric cancer: A potential target for gene therapy. Oncotarget 2015; 6:19685-19705.
26. Caiazza C, Mallardo M. The roles of miR-25 and its targeted genes in development of human cancer. Microrna 2016; 5:113-119.
27. Yuan DD, Zhu ZX, Zhang X, Liu J. Targeted therapy for gastric cancer: Current status and future directions (Review). Oncol Rep 2016; 35:1245-1254.
28. Feng F, Liu J, Wang F, Zheng G, Wang Q, Liu S, et al. Prognostic value of differentiation status in gastric cancer. BMC Cancer 2018; 18:865-870.
29. Chen HT, Liu H, Mao MJ, Tan Y, Mo XQ, Meng XJ, et al. Crosstalk between autophagy and epithelial-mesenchymal transition and its application in cancer therapy. Mol Cancer 2019; 18:101-119.
30. Yue B, Song C, Yang L, Cui R, Cheng X, Zhang Z, et al. METTL3-mediated N6-methyladenosine modification is critical for epithelial-mesenchymal transition and metastasis of gastric cancer. Mol Cancer 2019; 18:142-156.
31. Li W, Li S, Deng L, Yang S, Li M, Long S, et al. Decreased MT1-MMP in gastric cancer suppressed cell migration and invasion via regulating MMPs and EMT. Tumour Biol 2015; 36:6883-6889.
32. Dube DH, Bertozzi CR. Glycans in cancer and inflammation--potential for therapeutics and diagnostics. Nat Rev Drug Discov 2005; 4:477-488.
33. Guo H, Nairn A, dela Rosa M, Nagy T, Zhao S, Moremen K, et al. Transcriptional regulation of the protocadherin β cluster during Her-2 protein-induced mammary tumorigenesis results from altered N-glycan branching. J Biol Chem 2012; 287:24941-24954.
34. Wang C, Yang Y, Yang Z, Liu M, Li Z, Sun L, et al. EGF-mediated migration signaling activated by N-acetylglucosaminyltransferase-V via receptor protein tyrosine phosphatase kappa. Arch Biochem Biophys 2009; 486:64-72.