文章摘要

MiR-466对胃癌细胞增殖和迁移的影响及机制

作者: 1王佳春, 1全毅红
1 华中科技大学同济医学院附属武汉中心医院中医科,武汉 43003
通讯: 全毅红 Email: yihongquan846@126.com
DOI: 10.3978/j.issn.2095-6959.2019.12.003

摘要

目的:研究miR-466对胃癌细胞系增殖和迁移的影响,并阐明其可能的机制。方法:实时聚合酶链反应 (real-time polymerase chain reaction,real-time PCR)测定胃癌细胞系AGS,BSG823,MGC-803,Hs746T及正常胃黏膜上皮细胞系GES-1中miR-466的表达。将胃癌细胞系Hs746T分成3组:miR-466过表达组(miR-466组)、阴性对照组(miR-NC组)及空白对照组(Mock组)。用LipofectamineTM 3000分别转染miR-466 mimic,miR-NC序列,Mock组为空白对照。细胞计数试剂盒-8(cell counting kit-8,CCK-8)法和细胞划痕实验分别测定3组细胞增殖和迁移能力,蛋白质印迹测定3组Runt相关转录因子2(Runt-related transcription factor 2,RUNX2)蛋白表达。结果:miR-466在胃癌细胞系AGS,BSG823,MGC-803和Hs746T中的表达量低于正常胃黏膜上皮细胞系GES-1 (P<0.05)。转染0,24,48 h,OD450 nm值3组差异无统计学意义(P>0.05);转染后72及96 h,miR-466组OD450 nm低于miR-NC组和Mock组(P<0.05)。miR-466组划痕愈合率低于miR-NC组[(19.7±6.8)% vs (69.3±7.8)%,P<0.01],miR-NC组与Mock组划痕愈合率差异无统计学意义(P>0.05)。miR-466组RUNX2蛋白相对表达量低于miR-NC组(0.38±0.04 vs 1.00±0.03,P<0.05);miR-NC组与Mock组RUNX2蛋白表达量差异无统计学意义(P>0.05)。结论:在胃癌细胞系中miR-466低表达,上调miR-466表达可抑制胃癌细胞增殖和迁移,其机制可能与RUNX2蛋白下调表达有关。
关键词: miR-466;胃癌;增殖;迁移;Runt相关转录因子2

Effect of miR-466 on the proliferation and migration of gastric carcinoma cell line and its mechanism

Authors: 1WANG Jiachun, 1QUAN Yihong
1 Department of Traditional Chinese Medicine, Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

CorrespondingAuthor: QUAN Yihong Email: yihongquan846@126.com

DOI: 10.3978/j.issn.2095-6959.2019.12.003

Abstract

Objective: To investigate the effect of miR-466 on the proliferation and migration of gastric carcinoma cell lines, and explain its mechanism. Methods: Real-time polymerase chain reaction (real-time PCR) was used to detect the expression of miR-466 in normal gastric cell line, GES-1 and gastric cancer cell lines, AGS, BSG823, MGC-803 and Hs746T. The Hs746T cell line was assigned into three groups: miR-466 over-expression group (miR-466 group), negative control group (miR-NC group) and blank control group (Mock group), which was transfect with miR-466 mimic, miR-NC sequence and phosphate buffered saline with Tween-20 (PBST) by LipofectamineTM 3000. The proliferation and migration ability were analyzed using Cell Counting Kit-8 (CCK-8) and wound scratch assay between three groups. The expression of Runt-domain containing protein (RUNX2) protein was analyzed by Western blot between three groups. Results: The expression of miR-466 in four gastric carcinoma cell lines, AGS, BSG823, MGC-803 and Hs746T, was significantly lower than that in normal gastric cell line, GES-1 (P<0.05). There was no significant difference in OD450 nm value between the three groups at 0, 24 and 48 h after transfection (P>0.05). The OD450 nm value of miR-466 group was significantly lower than that in miR-NC group and Mock group after transfecting for 72 and 96 h, respectively (P<0.05). Scar healing rate of miR-466 group was significantly lower than that in miR-NC group [(19.7±6.8)% vs (69.3±7.8)%, P<0.01]. There was no significant difference in scratch healing rate between miR-NC group and Mock group (P>0.05). The expression level of RUNX2 in miR-466 group was significantly lower than in miR-NC group (0.38±0.04 vs 1.00±0.03, P<0.05). There was no significant difference in the expression of RUNX2 protein between miR-NC group and Mock group (P>0.05). Conclusion: The expression of miR-466 in gastric cancer cell lines is significantly decreased. Over-expression of miR-466 could inhibit the proliferation and migration of gastric cancer cells, probably by targeting RUNX2.
Keywords: miR-466; gastric carcinoma; proliferation; migration; Runt-domain containing protein 2