Yuanhang Li1, Weijun Bai2, Jianjun Zhang2. 1. Cancer Hospital of China Medical University, No. 44 Xiaoheyan Road, Dadong District, Shenyang 110042, Liaoning Province, PR China; Liaoning Cancer Hospital & Insititute, No. 44 Xiaoheyan Road, Dadong District, Shenyang 110042, Liaoning Province, PR China. Electronic address: yamindeng@163.com. 2. Cancer Hospital of China Medical University, No. 44 Xiaoheyan Road, Dadong District, Shenyang 110042, Liaoning Province, PR China; Liaoning Cancer Hospital & Insititute, No. 44 Xiaoheyan Road, Dadong District, Shenyang 110042, Liaoning Province, PR China.
Abstract
OBJECTIVE: To explore the biological functions of miR-200c-5p/MAD2L1 axis on the proliferation and metastasis of human hepatocellular carcinoma (HCC) cells. METHODS: The expression levels of miR-200c-5p and MAD2L1 in HCC tissues, adjacent tissues as well as HCC cell lines were detected by RT-qPCR or Western blot. The interaction between miR-200c-5p and MAD2L1 was verified by dual luciferase reporter gene system. Transfection was performed to manipulate the expression of miR-200c-5p and MAD2L1 in HCCLM3 cells. Colony formation, MTT, wound healing and Transwell assays were applied to measure the cell proliferation, migration and invasion of HCC, besides, flow cytometry analysis was also conducted to evaluate HCC cell cycle and apoptosis. RESULTS: Low expression of miR-200c-5p and remarkable overexpression of MAD2L1 was uncovered in HCC tissues and cells compared with the normal. The aberrant expression of miR-200c-5p and MAD2L1 was correlated with tumor stage, adjacent organ invasion and prognosis. Direct target relationship between miR-200c-5p and MAD2L1 was confirmed by dual luciferase reporting assay. Up-regulation of miR-200c-5p downregulated MAD2L1 and suppressed the proliferation, migration, invasion and induced apoptosis and cell cycle arrest of HCC cells. Moreover, MAD2L1 promoted HCC cell viabilities and co-transfection of MAD2L1 restored the anti-tumor effects of miR-200c-5p overexpression. CONCLUSION: Replenishing of miR-200c-5p inhibited the proliferation, migration and invasion of HCC cells by suppressing MAD2L1. MiR-200c-5p can serve as a prognostic indicator and a promising therapeutic target for HCC patients.
OBJECTIVE: To explore the biological functions of miR-200c-5p/MAD2L1 axis on the proliferation and metastasis of humanhepatocellular carcinoma (HCC) cells. METHODS: The expression levels of miR-200c-5p and MAD2L1 in HCC tissues, adjacent tissues as well as HCC cell lines were detected by RT-qPCR or Western blot. The interaction between miR-200c-5p and MAD2L1 was verified by dual luciferase reporter gene system. Transfection was performed to manipulate the expression of miR-200c-5p and MAD2L1 in HCCLM3 cells. Colony formation, MTT, wound healing and Transwell assays were applied to measure the cell proliferation, migration and invasion of HCC, besides, flow cytometry analysis was also conducted to evaluate HCC cell cycle and apoptosis. RESULTS: Low expression of miR-200c-5p and remarkable overexpression of MAD2L1 was uncovered in HCC tissues and cells compared with the normal. The aberrant expression of miR-200c-5p and MAD2L1 was correlated with tumor stage, adjacent organ invasion and prognosis. Direct target relationship between miR-200c-5p and MAD2L1 was confirmed by dual luciferase reporting assay. Up-regulation of miR-200c-5p downregulated MAD2L1 and suppressed the proliferation, migration, invasion and induced apoptosis and cell cycle arrest of HCC cells. Moreover, MAD2L1 promoted HCC cell viabilities and co-transfection of MAD2L1 restored the anti-tumor effects of miR-200c-5p overexpression. CONCLUSION: Replenishing of miR-200c-5p inhibited the proliferation, migration and invasion of HCC cells by suppressing MAD2L1. MiR-200c-5p can serve as a prognostic indicator and a promising therapeutic target for HCC patients.
Authors: Xianyu Hu; Suwen Bai; Lingyi Li; Pengfei Tian; Sun Wang; Ning Zhang; Bing Shen; Juan Du; Shengxiu Liu Journal: Exp Biol Med (Maywood) Date: 2021-01-20