Experimental study on inhibition of migration and invasion of lung cancer cells by miR?144?3p targeting ONECUT2

LU Zhansheng; LIU Bo; SUN Yongpan; KONG Demiao; YAO Jing; ZHOU Xiaojiang; XU Chuan; LUO Meng; ZHANG Qingbin

doi:10.3969/j.issn.1009-0460.2019.09.002

Abstract：Objective To analyze the effect of miR?144?3p on the migration and invasion of lung cancer cells and explore its possible mechanism.Real?time fluorescence quantitative polymerase chain reaction (QPCR) was used to detect the expression of miR?144?3p and ONECUT2 in lung cancer NCI?H460 cells and normal bronchial epithelial cells HBE.miR?144?3p mimics ( miR?144?3p group), miR?NC (miR?NC group), si?con ( si?con group), si?ONECUT2 ( si?ONECUT2 group), miR?144?3p mimics and pcDNA (miR?144?3p+pcDNA group), miR?144?3p mimics and pcDNA?ONECUT2 (miR?144?3p+pcDNA?ONECUT2 group) were transfected into NCI?H460 cells by liposome method.Transwell assay was used to detect the migration and invasion ability of cells in each group. The protein expression of ONECUT2, E?cadherin, N?cadherin, and Vimentin of each group were detected by Western blotting.Dual lu?ciferase reporter gene assay was used to detect the targeting relationship between miR?144?3p and ONECUT2. Results Compared with HBE group, the expression of miR?144?3p and ONECUT2 in NCI?H460 group were significantly decreased and increased ( P＜0. 05). Overexpression of miR?144?3p or silence of ONECUT2 could significantly inhibit the migration and invasion of NCI?H460 cells, up?reg?ulate E?cadherin protein expression, down?regulate N?cadherin and Vimentin protein expression.MiR?144?3p inhibited the luciferase ac?tivity of wild?type ONECUT2 3’?UTR reporter gene vector, but had no effect on the luciferase activity of mutant ONECUT2 3’?UTR?MUT.Overexpression of ONECUT2 reversed the expression of EMT?related protein in NCI?H460 cells by miR?144?3p. Conclusion Overexpression of miR?144?3p can inhibit the migration and invasion of lung cancer cells. The mechanism may be related to the inhibi?tion of EMT by targeting ONECUT2, which can provide a reference for the treatment of lung cancer.