MicroRNA-1 inhibits proliferation of hepatocarcinoma cells by targeting endothelin-1

• Published in: Life sciences (1973) Vol. 91; no. 11-12; pp. 440 - 447
• Main Authors: Li, Dong, Yang, Pengyuan, Li, Hua, Cheng, Peng, Zhang, Ling, Wei, Dong, Su, Xiaomei, Peng, Jingjing, Gao, Hui, Tan, Yong, Zhao, Zhenguo, Li, Yan, Qi, Zhongchun, Rui, Yaocheng, Zhang, Tao
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 05.10.2012
• Subjects: binding sites; bioinformatics; Blotting, Western; carcinogenesis; Carcinoma, Hepatocellular - drug therapy; Carcinoma, Hepatocellular - physiopathology; Cell Proliferation - drug effects; Cellular proliferation; Endothelin-1; Endothelin-1 - antagonists & inhibitors; Endothelin-1 - physiology; gene overexpression; genes; Hep G2 Cells - drug effects; Hepatocellular carcinoma; hepatoma; Humans; Liver Neoplasms - drug therapy; Liver Neoplasms - physiopathology; luciferase; Luciferases; messenger RNA; microRNA; MicroRNAs; MicroRNAs - therapeutic use; plasmids; quantitative polymerase chain reaction; Real-Time Polymerase Chain Reaction; transfection; Western blotting; Hepatocellular carcinoma; Cellular proliferation; MicroRNAs; Endothelin-1
• ISSN: 0024-3205; 1879-0631
• DOI: 10.1016/j.lfs.2012.08.015

MicroRNA-1 (miR-1) has been demonstrated as a tumor-suppressive miRNA, which shows a down-regulated pattern in several human malignancies including hepatocellular carcinoma (HCC). However, the pathophysiologic roles of miR-1 and their mechanisms in HCC tumorigenesis are still not totally elucidated. Pre-miR-1 was cloned into pSuper plasmid to overexpress the miR-1 in hepatoma cells. Real-time PCR and Western blot were applied to detect miR-1, ET-1 mRNA and protein levels respectively. Dual luciferase reporter assay was conducted to investigate the binding site of miR-1 on 3′UTR of ET-1 mRNA. Proliferation of hepatoma cells was evaluated by MTT assay. We observed that over-expression of miR-1 by miRNA-expressing plasmid transfection in HepG2 and Hep3B cells significantly reduced the proliferation of these cells. To explore the mechanism, we examined the potential target genes of miR-1 by bioinformatics. A potent mitogen, Endothelin-1 (ET-1), attracted our attention. Elevated expression of ET-1 but reduced miR-1 level was detected both in human liver cancer tissues and in hepatoma cell lines using Western Blot and miRNA real-time PCR respectively. By the over-expression and inhibition of miR-1 in HepG2 and Hep3B, we confirmed that miR-1 negatively regulated ET-1 expression in hepatoma cells. A luciferase reporter assay showed that miR-1 regulation was established by pairing to a complementary binding site within the ET-1 3′UTR. Finally, attenuated proliferation of hepatoma cells by over-expression of miR-1 could be partially restored by exogenous ET-1 treatment. Our findings demonstrate that miR-1 could inhibit ET-1 expression to attenuate the proliferation of hepatoma cells.