Effect of MicroRNA-31 on Migration and Invasion of AGS Cells of Gastric Cancer and on The Expression Change of LRH-1_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

 GAOHong-yuan ¹ , TIANShou-jin ² ,  WUXue-yong ¹

1. Department of Oncology, Jing'an District Centre Hospital of Shanghai, Huashan Hospital, Jing'an Branch of Fudan University, Shanghai 200041, China;
2. Department of Gastric Surgery, Affiliated Zhucheng People's Hospital of Weifang Medical University, Zhucheng 262200, Shandong Province, China;

Corresponding author：

WUXue-yong, Email: Wuxy6309@126.com

Keywords：

MicroRNA-31; Gastric cancer; Migration; Proliferation; Liver receptor homolog

DOI：

10.7507/1007-9424.20140164

Video：

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Objective To investigate the effect of up-regulation of microRNA-31(miR-31) on the biological behaviour in AGS cell of gastric cancer and on the expression of liver receptor homolog-1(LRH-1), and to analyze the possible mechanisms of miR-31 on initiation and development of gastric cancer. Methods AGS cells were divided into 3 groups, receiving miR-31 transfection(MT group), empty liposomes transfection(NC group), and treatment of PBS (BC group). Then the cells' proliferation was determined by cell counting kit-8(CCK-8), the apoptosis situation was determined by flow cytometer, the migration was determined by Transwell test, the expression of LRH-1 protein was tested by Western blot method, and the target of miR-31 was tested by luciferase reporter assay. Results The cell's proliferation results showed that the mean of A₄₅₀ value in MT, NC, and BC groups were 1.31, 2.26, and 2.14 respectively on the 4 days after transfection, which lower in MT group(P<0.01).Results of flow cytometer experiment showed that the mean of apoptosis ratio of MT, NC, and BC groups were 39.5%, 9.3%, and 10.0% respectively, the mean of proportion of cell in G₁+S stage were 92.54%, 73.23%, and 74.58% respectively, which both lower in MT group (P<0.05).Results of Transwell experiment showed that the mean of number of migrated cells in MT group was lower (P<0.05).Results of Western blot experiment demonstrated that the expression level of LRH-1 protein in MT group was lower than those of BC group and NC group(P<0.01). Conclusions Up-regulation of miR-31 can obviously inhibit the proliferation of AGS cell, promoting its apoptosis and depressing its migration ability. On the other side, the up-regulation of miR-31 can also inhibit the expression level of LRH-1 protein, which indirectly induces the inhibition of proliferation of AGS cell. So miR-31 may be an important regulator in the initiation and development of gastric cancer through regulating LRH-1 gene.

Citation： GAOHong-yuan, TIANShou-jin, WUXue-yong. Effect of MicroRNA-31 on Migration and Invasion of AGS Cells of Gastric Cancer and on The Expression Change of LRH-1. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2014, 21(6): 682-687. doi: 10.7507/1007-9424.20140164 Copy

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1. Sundermeier TR, Palczewski K.The physiological impact of microRNA gene regulation in the retina[J].Cell Mol Life Sci, 2012, 69(16):2739-2750.
2. Inui M, Martello G, Piccolo S.MicroRNA control of signal transduction[J].Nat Rev Mol Cell Biol, 2010, 11(4):252-263.
3. Li JY, Yong TY, Michael MZ, et al.Review:the role of microRNAs in kidney disease[J].Nephrology (Carlton), 2010, 15(6):599-608.
4. Bhatt K, Mi QS, Dong Z.MicroRNAs in kidneys:biogenesis, regulation, and pathophysiological roles[J].Am J Physiol Renal Physiol, 2011, 300(3):F602-F610.
5. Gommans WM, Berezikov E.Controlling miRNA regulation in disease[J].Methods Mol Biol, 2012, 822:1-18.
6. Ragusa M, Caltabiano R, Russo A, et al.MicroRNAs in vitreus humor from patients with ocular diseases[J].Mol Vis, 2013, 19:430-440.
7. Zhang Y, Guo J, Li D, et al.Down-regulation of miR-31 expression in gastric cancer tissues and its clinical significance[J].Med Oncol, 2010, 27(3):685-689.
8. Valastyan S, Reinhardt F, Benaich N, et al.A pleiotropically acting microRNA, miR-31, inhibits breast cancer metastasis[J].Cell, 2009, 137(6):1032-1046.
9. Xi S, Yang M, Tao Y, et al.Cigarette smoke induces C/EBP-β-mediated activation of miR-31 in normal human respiratory epithelia and lung cancer cells[J].PLoS One, 2010, 5(10):e13764.
10. Creighton CJ, Fountain MD, Yu Z, et al.Molecular profiling uncovers a p53-associated role for microRNA-31 in inhibiting the proliferation of serous ovarian carcinomas and other cancers[J].Cancer Res, 2010, 70(5):1906-1915.
11. Wszolek MF, Rieger-Christ KM, Kenney PA, et al.A MicroRNA expression profile defining the invasive bladder tumor phenotype[J].Urol Oncol, 2011, 29(6):794-801.
12. Wang SL, Zheng DZ, Lan FH, et al.Increased expression of hLRH-1 in human gastric cancer and its implication in tumorigenesis[J].Mol Cell Biochem, 2008, 308(1-2):93-100.
13. Chand AL, Herridge KA, Thompson EW, et al.The orphan nuclear receptor LRH-1 promotes breast cancer motility and invasion[J].Endocr Relat Cancer, 2010, 17(4):965-975.
14. Target Scan[DB/OL].http://www.targetscan.org/cgi-bin/targetscan/vert_50/targetscan.cgi?mirg=hsa-miR-31.
15. Pic Tar[DB/OL].http://pictar.mdc-berlin.de/cgi-bin/PicTar_vertebrate.cgi?action=Search%20for%20targets%20of%20a%20miRNA&name2=hsa-miR-31.
16. DIANA LAB[DB/OL].http://diana.cslab.ece.ntua.gr/DianaTools/index.php?r=microtv4/results&threshold=0.3&mirnas=MIMAT0000089.
17. Barranco SC, Townsend CM Jr, Casartelli C, et al.Establishment and characterization of an in vitro model system for human adenocarcinoma of the stomach[J].Cancer Res, 1983, 43(4):1703-1709.
18. 庄亚琼,李建英,陈治新,等.丁酸钠对胃癌细胞株AGS增殖分化的影响及其机制[J].癌症, 2008, 27(8):828-834.
19. Gunassekaran GR, Priya DK, Gayathri R, et al.In vitro and in vivo studies on antitumor effects of gossypol on human stomach adenocarcinoma (AGS) cell line and MNNG induced experimental gastric cancer[J].Biochem Biophys Res Commun, 2011, 411(4):661-666.
20. Chung CP, Hsia SM, Lee MY, et al.Gastroprotective activities of adlay (Coix lachryma-jobi L.var.ma-yuen Stapf) on the growth of the stomach cancer AGS cell line and indomethacininduced gastric ulcers[J].J Agric Food Chem, 2011, 59(11):6025-6033.
21. Zheng Q, Zhao LY, Kong Y, et al.CDK-associated Cullin 1 can promote cell proliferation and inhibit cisplatin-induced apoptosis in the AGS gastric cancer cell line[J].World J Surg Oncol, 2013, 11:5-16.
22. 周海燕,王宽松,胡忠良,等.miR-193b对胃癌细胞浸润和转移的影响[J].中国普通外科杂志, 2011, 20(4):377-382.
23. Schoonjans K, Dubuquoy L, Mebis J, et al.Liver receptor homolog 1 contributes to intestinal tumor formation through effects on cell cycle and inflammation[J].Proc Natl Acad Sci U S A, 2005, 102(6):2058-2062.
24. Botrugno OA, Fayard E, Annicotte JS, et al.Synergy between LRH-1 and beta-catenin induces G1 cyclin-mediated cell proliferation[J].Mol Cell, 2004, 15(4):499-509.
25. Miki Y, Clyne CD, Suzuki T, et al.Immunolocalization of liver receptor homologue-1(LRH-1) in human breast carcinoma:possible regulator of insitu steroidogenesis[J].Cancer Lett, 2006, 244(1):24-33.
26. Annicotte JS, Chavey C, Servant N, et al.The nuclear receptor liver receptor homolog-1 is an estrogen receptor target gene[J].Oncogene, 2005, 24(55):8167-8175.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Effect of MicroRNA-31 on Migration and Invasion of AGS Cells of Gastric Cancer and on The Expression Change of LRH-1

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