miR-144通过LIS1抑制胆管癌细胞的增殖和侵袭

miR-144 inhibits the proliferation and invasion of cholangiocarcinoma cells through LIS1

目的:筛选并验证miR-144的可能作用靶点,阐明miR-144的生物学功能。方法:通过miRanda、PicTar和Target Scan软件预测并验证miR-144可能的靶基因;构建双荧光素酶报告基因,验证miR-144与靶基因的直接结合,并明确其结合位点;利用siRNA干扰沉默无脑回致病基因LIS1后检测LIS1对胆管癌细胞增殖、迁移和侵袭能力的影响,Western Blot验证LIS1对增殖和侵袭相关的信号通路的影响,验证miR-144与LIS1功能学上的一致性。结果:LIS1是miR-144可能的靶基因之一;miR-144可以通过结合于LIS1的3′UTR两个可能的结合位点调控LIS1的表达;干扰LIS1后可以下调p-AKT和MMP2的水平,从而影响胆管癌细胞的增殖和侵袭。结论:miR-144通过直接作用于LIS1影响胆管癌细胞的增殖和侵袭能力,这种作用是通过下调p-AKT和MMP2来实现的。

Objective:To screen and validate the potential targets of miR-144,and then further clarify the biological function of miR-144.Methods:The potential target genes of miR-144 were predicted and verified by miRanda,PicTar and Target Scan softwares.The dual luciferase report system was established to validate the direct combination of miR-144 to target genes and clarify their binding sites.The effect of LIS1 on proliferation,migration,and invasion ability of cholangiocarcinoma cells was detected by using specific siRNA targeted LIS1,and Western Blot was used to detect the influence of LIS1 on signaling pathways related to proliferation,migration,and invasion,to validate the functional consistency of miR-144 and LIS1.Results:LIS1 was one of the potential target genes of miR-144;miR-144 could combine with LIS1 at two sites of 3′UTR to regulate LIS1 expression;knockdown of LIS1 could reduce the protein levels of p-AKT and MMP2,which affects the proliferation and invasion of cholangiocarcinoma cells.Conclusion:miR-144 decreases the expression of p-AKT and MMP2 after binding to LIS1 directly,which influences the proliferation and invasion ability of cholangiocarcinoma cells eventually.