摘要
目的观察miR-630在胃癌组织中的表达变化,及miR-630对胃癌细胞株SGC-7901增殖、侵袭能力的影响。方法收集131例胃癌患者的胃癌组织及癌旁正常胃组织,采用实时荧光定量PCR法检测miR-630。培养SGC-7901细胞,分为实验组、空载组和对照组,实验组转染miR-630,空载组转染空质粒,对照组仅加入PBS;分别于培养12、24、48、72 h后采用MTT法检测细胞增殖能力;培养24 h后以Transwell小室检测细胞侵袭能力;培养24 h后,采用Western blotting法检测细胞中的p27、MMP-2、MMP-9蛋白。结果胃癌组织、癌旁组织中miR-630的相对表达量分别为1.93±0.41、2.74±0.48,胃癌组织中miR-630相对表达量低于癌旁组织(P均<0.05)。肿瘤直径≥5 cm者胃癌组织中miR-630相对表达量低于直径<5 cm者,有淋巴结转移者低于无淋巴结转移者,浸润深度为T2~T4级者低于T1级者,Ⅲ~Ⅳ期者低于Ⅰ~Ⅱ期者(P均<0.05)。转染24、48、72 h后,实验组细胞增殖能力均低于转染同时点的空载组和对照组(P均<0.01)。实验组、空载组、对照组穿膜细胞数分别为15.4±2.7、24.3±23.1、29.7±19.4,实验组穿膜细胞数少于对照组和空载组(P均<0.01)。实验组细胞中p27蛋白相对表达量高于空载组和对照组,MMP-2、MMP-9蛋白相对表达量低于空载组和对照组(P均<0.01)。结论胃癌组织中miR-630表达减少,mir-630低表达可能与胃癌的发生发展有关;miR-630有抑制胃癌细胞增殖、侵袭的作用,机制可能与p27蛋白表达上调及MMP-2、MMP-9蛋白表达下调有关。
Objective To investigate the expression of miR-630 in gastric carcinoma and its effect on the proliferation and invasion ability of gastric cancer cell line SGC-7901 in vitro. Methods The gastric cancer tissues and adjacent normal gastric tissues were collected,from 131 patients with gastric cancer. The expression of miR-630 in the tissues was detected by RT-PCR. The cultured SGC-7901 cells were divided into 3 groups ： the experimental group, empty vector group and control group which were transfeeted with miR-630, empty vector and PBS, respectively. The cells were cultured for 12, 24, 48 and 72 h, and MTY assay was employed to test the proliferation ability of cells. At 24 h, Transwell assay was used to test the invasion ability of cells, and Western blotting was used to investigate the expression of p27, MMP-2, MMP-9 in SGC-7901 cells. Results The relative expression levels of miR-630 in the gastric cancer tissues and adjacent normal tissues were 1.93 ± 0.41 and 2.74 ± 0.48, respectively. The relative expression of miR-630 in the gastric cancer tissues was lower than that in the adjacent normal tissues （ all P 〈 0.05）. The relative expression of miR-630 in gastric cancer patients with diameter ≥5 cm, with lymph node metastasis, in T2-T4 grade and in Ⅲ-Ⅳ phase was successively lower than that in patients with diameter 〉5 cm, without lymph node metastasis, in T2-T4 grade, and in Ⅲ-Ⅳ phase （all P 〈0.05）. In vitro studies, at 24, 48 and 72 h after transfection, the cell proliferation ability of the experimental group was lower than that of the empty vector group and control group （ all P 〈 0.01 ）. The transmembrane cells in the experimental group, empty vector group and control group were 15.4 ± 2.7, 24.3 ±23.1 and 29.7 ± 19.4, respectively. The number of the transmembrane cells in the experimental group was less than that in the empty vector group and the control group （ all P 〈 0.01 ）. The relative expression of p27 protein in the the experimental group was higher, and the expression of MMP-2 and MMP-9 protein was lower than that in the empty vector group and control group （ all P 〈 0.01 ）. Conclusions The expression of miR-630 in the gastric cancer tissues is decreased, and the low expression of mir-630 might be related to the development of gastric cancer. MiR-630 could inhibit the proliferation and invasion of gastric cancer cells, which may be related to the up-regulation of p27 and down-regulation of MMP-2 and MMP-9.
出处
《山东医药》
CAS
北大核心
2017年第15期9-12,共4页
Shandong Medical Journal
基金
甘肃省卫生行业科研计划资助项目(GSWST2013-02)
