LncRNA PINK1-AS调节miR-134-5p/SERPINE1轴对胃癌细胞增殖凋亡和上皮间质转化的影响

doi:10.3969/j.issn.1006-6233.2023.12.013

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摘要 目的: 探讨LncRNA PINK1-AS调节miR-134-5p/丝氨酸蛋白酶抑制剂分支E成员1(SERPINE1)轴对胃癌细胞增殖、凋亡和上皮间质转化(EMT)的影响。方法: qRT-PCR检测60例胃癌组织、癌旁组织中PINK1-AS、miR-134-5p、SERPINE mRNA的表达水平。体外培养胃癌细胞MKN28,将MKN28细胞分为control组、si-NC组、si-PINK1-AS组、pc-NC组、pc-PINK1-AS组、miR-NC组、miR-134-5p mimics组;qRT-PCR检测各转染组细胞PINK1-AS、miR-134-5p、SERPINE mRNA的表达水平,cck-8检测细胞增殖;流式细胞术检测细胞凋亡率;Transwell实验细胞迁移和侵袭;western blot检测细胞SERPINE1、Ki-67、cleaved caspase-3、E-cadherin、N-cadherin、vimentin蛋白表达水平。双荧光素酶报告基因实验检测LncRNA PINK1-AS和miR-134-5p、miR-134-5p和SERPINE1的靶向关系。结果: 胃癌组织中PINK1-AS和SERPINE mRNA的表达显著高于癌旁组织,miR-134-5p表达低于癌旁组织(P<0.05)。下调PINK1-AS表达可降低MKN28细胞中PINK1-AS、SERPINE mRNA表达、OD450值(24、48h)、细胞迁移数和侵袭数、SERPINE1、Ki-67、N-cadherin、vimentin蛋白表达,提高miR-134-5p、细胞凋亡率、cleaved caspase-3、E-cadherin蛋白表达(P<0.05);过表达PINK1-AS可提高MKN28细胞中PINK1-AS、SERPINE mRNA表达、OD450值(24、48h)、细胞迁移数和侵袭数、SERPINE1、Ki-67、N-cadherin、vimentin蛋白表达,降低miR-134-5p、凋亡率、cleaved caspase-3、E-cadherin蛋白表达(P<0.05);过表达miR-134-5p可提高MKN28细胞中miR-134-5p表达、细胞凋亡率、cleaved caspase-3、E-cadherin蛋白表达,降低SERPINE mRNA、OD450值(24、48h)、细胞迁移数和侵袭数、SERPINE1、Ki-67、N-cadherin、vimentin蛋白表达(P<0.05)。结论: LncRNA PINK1-AS可调控miR-134-5p/SERPINE1轴进而影响胃癌细胞增殖、凋亡和EMT。

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	李凡
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	冯浩

关键词 ：胃癌细胞, LncRNA PINK1-AS, miR-134-5p/SERPINE1轴, 上皮间质转化

Abstract：Objective: To investigate the effects of LncRNA PINK1-AS on the proliferation, apoptosis and epithelial mesenchymal transformation (EMT) of gastric cancer cells by regulating the miR-134-5p/serine protease inhibitor clade E member 1 (SERPINE1) axis. Methods: QRT-PCR was applied to detect the expression levels of PINK1-AS, miR-134-5p, and SERPINE mRNA in 60 cases of gastric cancer tissues and 60 cases of adjacent tissues. Gastric cancer cells MKN28 were cultured in vitro and grouped into control group, si-NC group, si-PINK1-AS group, pc-NC group, pc-PINK1-AS group, miR-NC group, and miR-134-5p mimics group; QRT-PCR was applied to detect the expression levels of PINK1-AS, miR-134-5p, and SERPINE mRNA in cells of each transfection group, Cck-8 was applied to detect cell proliferation; Flow cytometry was applied to detect cell apoptosis rate; Transwell test was applied to detect cell migration and invasion; Western blot was applied to detect the expression levels of SERPINE1, Ki-67, cleaved caspase-3, E-cadherin, N-cadherin, and vimentin proteins in cells. Double Luciferase reporter gene experiment was applied to detect the targeting relationship between PINK1-AS and miR-134-5p, and between miR-134-5p and LncRNA SERPINE1. Results: The expression of PINK1-AS and SERPINE mRNA in gastric cancer tissue was obviously higher than that in adjacent tissues, while the expression of miR-134-5p was lower than that in adjacent tissues (P<0.05). Down-regulation the expression of PINK1-AS can reduce the PINK1-AS, SERPINE mRNA, OD450 values (24, 48h), cell migration and invasion, the expression of SERPINE1, Ki-67, N-cadherin, vimentin proteins in MKN28 cells ,increased the expression of miR-134-5p, cell apoptosis rate, the expression of cleaved caspase-3, and E-cadherin proteins (P<0.05). Overexpression of PINK1-AS can increase the PINK1-AS, SERPINE mRNA, OD450 values (24, 48h), cell migration and invasion, the expression of SERPINE1, Ki-67, N-cadherin, vimentin proteins in MKN28 cells, the expression of miR-134-5p, cell apoptosis rate, the expression of cleaved caspase-3, and E-cadherin proteins obviously decreased (P<0.05). Overexpression of miR-134-5p increased miR-134-5p expression, apoptosis rate, cleaved caspase-3, and E-cadherin protein expression, and decreased SERPINE mRNA, OD450 value (24, 48h), cell migration and invasion number, SERPINE1, Ki-67, N -cadherin, and vimentin protein expression (P<0.05). Conclusion: LncRNA PINK1-AS can regulate the miR-134-5p/SERPINE1 axis, thereby affecting proliferation, apoptosis and EMT of gastric cancer cells.

Key words： Gastric cancer cells LncRNA PINK1-AS miR-134-5p/SERPINE1 axis Epithelial mesenchymal transformation

基金资助:河北省医学科学研究课题,(编号:20220437)

通讯作者: 冯浩

引用本文:

李凡, 宋玉涛, 王峰, 冯浩. LncRNA PINK1-AS调节miR-134-5p/SERPINE1轴对胃癌细胞增殖凋亡和上皮间质转化的影响[J]. 河北医学, 2023, 29(12): 2004-2011.
LI Fan, SONG Yutao, WANG Feng, et al. Effects of LncRNA PINK1-AS on the Proliferation Apoptosis and Epithelial Mesenchymal Transformation of Gastric Cancer Cells by Regulating the miR-134-5p/SERPINE1 Axis. HeBei Med, 2023, 29(12): 2004-2011.

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http://www.hbyxzzs.cn/CN/10.3969/j.issn.1006-6233.2023.12.013 或 http://www.hbyxzzs.cn/CN/Y2023/V29/I12/2004

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