Circ_0000212靶向miR-637调控肺癌H1299细胞的增殖、凋亡和放射敏感性

Circ_0000212 Targets MiR-637 to Regulate Proliferation,Apoptosisand Radiosensitivity of Lung Cancer H1299 Cells

目的探讨环状RNA 0000212(Circular RNA 0000212,circ_0000212)对肺癌H1299细胞增殖、凋亡和放射敏感性的影响及其分子机制。方法选取本院2017年1月至2019年1月的30例肺癌组织及相应癌旁组织;体外培养肺癌H1299细胞,将其分为si-NC组、si-circ_0000212组、miR-NC组、miR-637组、si-circ_0000212+anti-miR-NC组、si-circ_0000212+anti-miR-637组;实时荧光定量PCR(reverse transcription real-time fluorescence quantitative PCR,RT-qPCR)检测circ_0000212和miR-637的表达水平;分别进行噻唑蓝(methylthiazolyldiphenyl-tetrazolium bromide,MTT)、流式细胞术、克隆形成实验和蛋白免疫印迹法(Western blot)检测细胞增殖、凋亡、放射敏感性和相关蛋白表达;双荧光素酶报告实验检测circ_0000212和miR-637的靶向关系。结果与癌旁组织比较,肺癌组织中circ_0000212和miR-637的表达水平分别显著升高和降低(P<0.05)。抑制circ_0000212表达或过表达miR-637均显著抑制H1299细胞的增殖,促进细胞凋亡和提高放射敏感性(P<0.05)。circ_0000212靶向调控miR-637表达(P<0.05)。且下调miR-637反转抑制circ_0000212表达对肺癌H1299细胞增殖、凋亡和放射敏感性的影响(P<0.05)。结论抑制circ_0000212表达可能通过上调miR-637调控肺癌H1299细胞的增殖、凋亡和放射敏感性。

Objective To investigate effects of circular RNA 0000212(Circular RNA 0000212,circ_0000212)on proliferation,apoptosis and radiosensitivity of lung cancer H1299 cells and underlying molecular mechanism.Methods 30 cases of lung cancer tissues and corresponding paracancerous tissues collected from January,2017 to January,2019 in our hospital were selected for the study.Lung cancer H1299 cells were cultured in vitro and divided into si-NC group,si-circ_0000212 group,miR-NC group,miR-637 group,si-circ_0000212+anti-miR-NC group,and si-circ_0000212+anti-miR-637 group.RT-qPCR was conducted to detect the expressions of circ_0000212 and miR-637.Methylthiazolyldiphenyl-tetrazolium bromide(MTT),flow cytometry,clonal formation assay and western blot were performed to measure cell proliferation,apoptosis,radiosensitivity and related protein expressions.Dual luciferase reporter assay was used to detect the targeting relationship between circ_0000212 and miR-637.Results Compared with paracancerous tissues,expression levels of circ_0000212 and miR-637 in lung cancer tissues were significantly increased and decreased,respectively(P<0.05).Inhibition of circ_0000212 expression or overexpression of miR-637 significantly inhibited H1299 cell proliferation,promoted cell apoptosis and enhanced radiosensitivity(P<0.05).circ_0000212 targeted miR-637 expression(P<0.05).In addition,a down-regulation of miR-637 reversed the effect of inhibiting the expression of circ_0000212 on the proliferation,apoptosis and radiosensitivity of lung cancer H1299 cells(P<0.05).Conclusion Inhibition of circ_0000212 expression may regulate proliferation,apoptosis and radiosensitivity of lung cancer H1299 cells by upregulating miR-637.