MicroRNA-129通过靶向调控HMGA2抑制胃癌细胞增殖、侵袭

摘要

Background:More and more evidences suggest that microRNA plays an important role in the development of gastric cancer (GC).Expression of miR-129 in GC tissues is found abnormal, however, the mechanism of miR-129 in cell proliferation and invasion is still undefined.Aims:To investigate the expression of miR-129 in GC tissue and GC cell lines and the mechanism of miR-129 in proliferation and invasion of SGC-7901 cells.Methods:Eighty-two GC tissues and corresponding paracancerous tissues were collected, human gastric epithelial cell line and different GC cell lines were cultured, qPCR was conducted to assess miR-129 expression.SGC-7901 cells were transfected with miR-129 mimic or miR-NC, and then were transfected with overexpressed HMGA2 plasmid.Colony formation assay was used to detect cell proliferation ability, and Transwell chamber was used to assess cell invasion ability.Pearson correlation analysis was used to analyze the correlation between miR-129 and HMGA2 mRNA expression.Luciferase assay was performed to determine the activity of luciferase.mRNA and protein expressions of miR-129, HMGA2 were determined by qPCR and Western blotting, respectively.Results:Compared with paracancerous tissues, expression of miR-129 was significantly decreased in GC tissues (P<0.05);when compared with human gastric epithelial cells, expression of miR-129 was significantly decreased in GC cell lines (P<0.05).Compared with miR-NC group, proliferation and invasion abilities of SGC-7901 cells were inhibited in miR-129 mimic group (P<0.05).HMGA2 mRNA expression in GC tissues was significantly upregulated (P<0.05), and was negatively correlated with miR-129 expression (r=-0.543 9, P<0.01).Luciferase activity in wild-type miR-129 mimic group was significantly lower than that in miR-NC group (P<0.05);mRNA and protein expressions of HMGA2 were decreased after transfection with miR-129 mimic (P<0.05).Compared with miR-129+vector group, proliferation and invasion of SGC-7901 cells were significantly increased in miR-129 mimic+HMGA2 group (P<0.05).Conclusions:The expression of miR-129 is decreased in GC tissue and cells;miR-129 inhibits SGC-7901 cells proliferation and invasion by negatively regulating HMGA2.%背景:越来越多的研究表明microRNA在胃癌发生、发展中起重要作用.有研究表明miR-129在胃癌组织中表达异常,但其对胃癌细胞增殖、侵袭的作用仍不明确.目的:探讨miR-129在胃癌组织和胃癌细胞株中的表达及其影响胃癌细胞增殖和侵袭的机制.方法:收集82例胃癌组织及其相应癌旁组织,培养人胃黏膜上皮细胞株和不同的胃癌细胞株,以qPCR法检测miR-129表达.将miR-129 mimic或miR-NC转染胃癌SGC-7901细胞后,转染HMGA2过表达质粒.以克隆形成实验观察细胞增殖情况,Transwell小室法检测细胞侵袭情况,Pearson相关分析评估miR-129表达与HMGA2表达的相关性,荧光素酶实验检测荧光素酶活性,qPCR和蛋白质印迹法分别检测miR-129、HMGA2 mRNA和蛋白表达.结果:与癌旁组织相比,胃癌组织中miR-129表达明显下降(P<0.05);与人胃黏膜上皮细胞株GES-1相比,各胃癌细胞株中miR-129表达明显下降(P<0.05).与miR-NC组相比,miR-129 mimic组SGC-7901细胞增殖、侵袭能力均明显下降(P<0.05).胃癌组织中HMGA2 mRNA表达明显增加(P<0.05),并与miR-129表达呈负相关(r=-0.543 9,P<0.01).野生型miR-129 mimic组荧光素酶活性显著低于miR-NC组(P<0.05);转染miR-129 mimic后HMGA2 mRNA和蛋白表达显著降低(P<0.05).与miR-129+阴性对照组相比,miR-129 mimic+HMGA2组细胞增殖、侵袭能力明显升高(P<0.05).结论:miR-129在胃癌组织和细胞中低表达,其可通过下调HMGA2抑制SGC-7901细胞的增殖和侵袭.