miR-605-3p/TRIB3/β-catenin信号通路调控肺癌的发展

miR-605-3p/TRIB3/β-catenin signaling pathway regulates lung cancer development

目的探讨miR-605-3p是否影响肺癌细胞A549的增殖和细胞活力。方法转染合成的miR-605-3p模拟物过表达miR-605-3p为过表达组,转染合成的miR-605-3p抑制剂敲低miR-605-3p为敲低组,通过CCK-8和MTT检测2组肺癌细胞A549的增殖水平和细胞活力。通过miRDB在线分析miR-605-3p潜在底物,并通过荧光素酶报告实验验证并分析潜在底物的作用。结果过表达组肺癌细胞A549的增殖水平和细胞活力较对照组均下降(t=12.45、15.38,P值均<0.05);敲低组肺癌细胞A549的增殖水平和细胞活力较对照组均上升(t=11.54、8.45,P值均<0.05)。miR-605-3p靶向TRIB3的3′端非编码区(t=17.32,P值均<0.05)。过表达TRIB3后,肺癌细胞A549的增殖水平和细胞活力均上升(t=10.45、9.04,P值均<0.05)。敲低TRIB3后,肺癌细胞A549的增殖水平和细胞活力均下降(t=8.43、17.43,P值均<0.05)。过表达miR-605-3p后,TRIB3的表达下降,β-catenin进入细胞核的量减少;敲低miR-605-3p后,TRIB3的表达上升,β-catenin进入细胞核的量增加。同时敲低miR-605-3p和TRIB3或miR-605-3p和β-catenin,发现肺癌细胞A549的增殖水平和细胞活力差异无统计学意义(t=0.42、0.61,P值均>0.05);过表达TRIB3的同时敲低β-catenin,发现肺癌细胞A549的增殖水平和细胞活力差异无统计学意义(t=0.56、0.21,P值均>0.05)。结论miR-605-3p靶向TRIB3抑制β-catenin的入核水平,抑制肺癌细胞A549的增殖水平和细胞活力。

Objective To investigate whether Mir-605-3P affects the proliferation and cell viability of lung cancer cell A549.Methods The overexpression of miR-605-3p mimics synthesized was treated as the overexpression group,while the knockdown of miR-605-3p inhibitor was treated as the knockdown group.The proliferation and cell viability of lung cancer cells A549 in the two groups were detected by CCK-8 and MTT.The potential substrates of miR-605-3P were analyzed online by miRDB,and the effects of the potential substrates were verified and analyzed by luciferase report experiment.Results After miR-605-3p was overexpressed,the proliferation level and cell viability of lung cancer cell A549 were decreased(t=12.45,15.38,both P<0.05);after miR-605-3p was knocked down,the proliferation level and cell viability of lung cancer cell A549 were increased(t=11.54,8.45,both P<0.05).miR-605-3p targets the 3'non-coding region of TRIB3(t=17.32,P<0.05).After over-expression of TRIB3,it was found that the proliferation level and cell viability of lung cancer cell A549 increased(t=10.45,9.04,both P<0.05).After knocking down TRIB3,it was found that the proliferation level and cell viability of lung cancer cell A549 were decreased(t=8.43,17.43,both P<0.05).After miR-605-3p was overexpressed,the expression of TRIB3 decreased,and the amount of β-catenin entering the nucleus decreased.After miR-605-3p was knocked down,the expression of TRIB3 increased,and the amount of β-catenin entering the nucleus increased.When miR-605-3p and TRIB3 or miR-605-3p and β-catenin were knocked down at the same time,the proliferation level and cell viability of lung cancer cell A549 were not significantly changed(t=0.42,0.61,both P>0.05).Overexpression of TRIB3 and knockdown of β-catenin simultaneously revealed no significant changes in the proliferation level and cell viability of lung cancer cell A549(t=0.56,0.21,both P>0.05).Conclusions miR-605-3p targets TRIB3 to inhibit the nuclear entry of β-catenin,and subsequently inhibits the proliferation and cell viability of lung cancer cell A549.