circ_0001955/miR-188-3p轴影响肺癌细胞增殖、迁移和凋亡的分子机制研究

Molecular Mechanism of Circ_0001955/miR-188-3p Axis Affecting Proliferation,Migration and Apoptosis of Lung Cancer Cells

目的探讨circ_0001955对肺癌细胞恶性行为的影响及可能机制。方法RT-qPCR检测41例肺癌组织及癌旁组织中circ_0001955和miR-188-3p表达,分析肺癌组织中circ_0001955和miR-188-3p表达的相关性。体外培养肺癌细胞A549,双荧光素酶报告基因实验验证A549细胞中circ_0001955和miR-188-3p调控关系。构建干扰si-circ_0001955或过表达miR-188-3p的A549细胞,用CCK-8法和克隆形成实验、划痕实验、流式细胞术及蛋白质印迹法分别检测细胞增殖、迁移、凋亡及细胞中Bax和Bcl-2蛋白表达。结果与癌旁组织比较,肺癌组织中circ_0001955表达升高(P<0.05),miR-188-3p表达降低(P<0.05)。肺癌组织中circ_0001955和miR-188-3p的表达呈负相关(r=-0.978,P<0.05)。circ_0001955可靶向结合miR-188-3p,且干扰circ_0001955促进A549细胞中miR-188-3p表达。干扰circ_0001955或过表达miR188-3p降低了A549细胞集落形成数、划痕愈合率及细胞中Bcl-2蛋白表达(P<0.05),提高了细胞增殖抑制率、凋亡率及Bax蛋白表达(P<0.05)。干扰miR-188-3p逆转干扰circ_0001955对A549细胞增殖、迁移和凋亡的影响。结论干扰circ_0001955可能通过上调miR-188-3p阻碍肺癌细胞增殖和迁移,并促进细胞凋亡。

Objective To investigate the effect of circ_0001955 on the malignant behavior of lung cancer cells and its possible mechanism.Methods RT-qPCR was used to detect the expression of circ_0001955 and miR-188-3p in 41 cases of lung cancer tissues and normal adjacent tissues.Pearson correlation analysis was used to analyze the correlation between circ_0001955 and miR-188-3p expression in lung cancer tissues.Lung cancer cells A549 was cultured in vitro,and the dual luciferase reporter gene experiment was performed to verify the regulatory relationship between circ_0001955 and miR-188-3p in A549 cells.A549 cells that were interfered with si-circ_0001955 or overexpressed miR-188-3p were constructed,and then CCK-8 method and clone formation test,scratch experiment,flow cytometry and Western blotting were used to detect cell proliferation,migration,apoptosis and the protein expression of Bax and Bcl-2.Results Compared with adjacent tissues,the expression of circ_0001955 in lung cancer tissue increased(P<0.05),but the expression of miR-188-3p decreased(P<0.05).The expression of circ_0001955 and miR-188-3p in lung cancer tissues was negatively correlated(r=-0.978,P<0.05).circ_0001955 could target miR-188-3p,and interference with circ_0001955 promoted the expression of miR-188-3p in A549 cells.circ_0001955 or overexpression of miR-188-3p decreased A549 cell colony formation,scratch healing rate and the protein expression of Bcl-2 in cells(P<0.05),while proliferation inhibition rate,apoptosis rate and the protein expression of Bax increased(P<0.05).Interference with miR-188-3p reversed the effect of interference with circ_0001955 on the proliferation,migration and apoptosis of A549 cells.Conclusion Interference with circ_0001955 may hinder the proliferation and migration ability of lung cancer cells and promote cell apoptosis by up-regulating miR188-3p.