miR—505抑制无脑回致病基因进而抑制肝内胆管细胞癌RBE细胞增殖和侵袭

miR-505 suppresses lissencephaly pathogenic gene,thereby inhibiting the proliferation and invasion of intrahepatic cholangiocarcinoma RBE cells

目的:观察miR-505对肝内胆管细胞癌(intrahepatic cholangiocarcinoma,ICC)细胞的增殖、S期阻滞和侵袭的影响,寻找miR-505在调控过程中的关键靶基因,阐明miR-505在肝内胆管细胞癌中的作用及其工作机制。方法:分析ICC组织内miR-505和LIS1表达变化,采用TargetScan和双荧光素酶报告基因试验证实LIS1是否为miR-505的靶基因。采用CCK8法、流式细胞法和Transwell实验分别检测miR-505和LIS1对RBE细胞增殖、S期细胞比值和侵袭的影响。蛋白质印迹法检测miR-505对MMP-2/p-AKT的影响。结果:miR-505在ICC组织中的表达降低,L1S1是miR-505靶基因,同时相比对照组,ICC组织中LIS1异常高表达。随后体外细胞实验发现转染模拟物能抑制RBE细胞的增殖,S期阻滞和侵袭,而LIS1能逆转这种抑制效果。过表达miR-505可以抑制MMP-2/p-AKT的表达。结论:miR-505表达的失调与ICC密切相关,并能通过直接靶向作用于LIS1抑制MMP-2/p-AKT的表达,从而参与调控ICC细胞的生物学行为,发挥抑制ICC的作用。

Objective:Observe the effects of miR-505 on intrahepatic cholangiocarcinoma(ICC)cell proliferation,S-phase arrest and invasion,find the key target gene of miR-505 in regulation process,and clarify the function and working mechanism of miR-505 in ICC.Methods:Analyzing the changes of miR-505 and LIS1 expression in ICC tissues,TargetScan and Dual luciferase reporter gene were adopted to verify whether LIS1 was the target gene of miR-505.CCK8,Flow cytometry and Transwell assays were used to respectively detect effects of miR-505 and LIS1 on RBE cell proliferation,S-phase cell ratio and invasion.Western blot was used to detect the effects of miR-505 on MMP-2/p-AKT.Results:miR-505 expression was decreased in ICC tissues,and LIS1 was the target gene of miR-505.Meanwhile,comparing with control group,LIS1 had highly abnormal expression in ICC tissues.Then,in vitro cell experiments found transinfecting miR-505 mimics could inhibit RBE cell proliferation,S-phase arrest and invasion,while LIS1 could reverse such inhibitory effects.Overexpression miR-505 could inhibit MMP-2/p-AKT expression.Conclusion:miR-505 expression had a close correlation with ICC,and inhibited MMP-2/p-AKT expression via directly targeting LIS1,thereby participating in the regulation of ICC cell biologic behaviors and playing an inhibitory role in ICC.