miR-142-5p靶向SOX5对胃癌细胞增殖、凋亡、迁移和侵袭的影响

Effects of miR-142-5p targeting SOX5 on proliferation、apoptosis、migration and invasion of gastric cancer cells

目的探讨微小RNA-142-5p(miR-142-5p)靶向Y染色体上的性别决定区相关高迁移率族盒蛋白-5(SOX5)对胃癌细胞BGC823的影响。方法通过qRT-PCR检测胃上皮细胞GES-1和胃癌细胞BGC823中miR-142-5p表达水平,MTT实验、流式细胞仪、Transwell实验分别检测miR-142-5p mimics组、NC(Negative control)组、对照组细胞增殖、凋亡、迁移和侵袭情况,Western-blot检测各组细胞SOX5、B淋巴细胞瘤-2基因相关X蛋白(Bax)、细胞周期蛋白D1(CyclinD1)、基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9)蛋白表达情况;双荧光素酶基因报告验证miR-142-5p与SOX5的关系。结果与胃上皮细胞GES-1比较,胃癌细胞BGC823中miR-142-5p表达水平下降(P<0.05)。而过表达miR-142-5p后,BGC823细胞增殖、迁移、侵袭能力均受到抑制,增殖相关蛋白CyclinD1、迁移侵袭相关蛋白MMP-2和MMP-9蛋白表达水平显著降低(P<0.05),细胞凋亡率显著升高(P<0.05),凋亡相关蛋白Bax表达水平显著升高(P<0.05),SOX5 mRNA和蛋白表达显著下降(P<0.05)。双荧光素酶实验证实miR-142-5p显著降低野生型SOX53′UTR萤光素酶活性,对突变型SOX53′UTR的萤光素酶活性无影响,由此证明miR-142-5p与SOX5存在靶向关系。结论miR-142-5p可能通过靶向SOX5来抑制BGC823细胞增殖、迁移及侵袭,促进细胞凋亡。

Objective To investigate the effect of microRNA-142-5p(miR-142-5p)targeting sex determining region Ybox protein 5(SOX5)on gastric cancer cell BGC823.Methods qRT-PCR was used to detect the expression level of miR-142-5p in gastric epithelial cells GES-1 and gastric cancer cells BGC823.The proliferation,apoptosis,migration and invasion of miR-142-5p mimics group,Negative control group and control group were detected by MTT assay,flow cytometry and Transwell test.Western blot was used to detect the expression of SOX5,Bcl2 associated X(Bax),CyclinD1,matrix metalloproteinase 2(MMP-2)and matrix metalloproteinase 9(MMP-9),and the relationship between miR-142-5p and SOX5 was verified by double luciferase gene report.Results Compared with GES-1 cells,the expression of miR-142-5p in BGC823 cells decreased(P<0.05),while the proliferation,migration and invasion of BGC823 cells were inhibited after overexpression of miR-142-5p.The expression levels of CyclinD1,MMP-2 and MMP-9 were significantly decreased(P<0.05),and the apoptosis rate was significantly increased(P<0.05).At the same time,double luciferase assay confirmed that miR-142-5p significantly decreased the luciferase activity of wild-type SOX53′UTR,but had no effect on the luciferase activity of mutant SOX53′UTR,which indicated that miR-142-5p had a targeted relationship with SOX5.Conclusion miR-142-5p may inhibit the proliferation,migration and invasion of BGC823 cells by targeting SOX5 and promoting apoptosis.