MiR-506 suppresses proliferation of hepatoma cells through targeting YAP mRNA 3′UTR
Abstract
Yue WANG1, Ming CUI2, Bao-di SUN2, Fa-bao LIU1, Xiao-dong ZHANG2, Li-hong YE1, *
1State Key Laboratory of Medicinal Chemical Biology, Department of Biochemistry, College of Life Sciences, Nankai University, Tianjin 300071, China; 2State Key Laboratory of Medicinal Chemical Biology, Department of Cancer Research, College of Life Sciences, Nankai University, Tianjin 300071, China
Aim: MiR-506 is a miRNA involved in carcinogenesis of several kinds of cancer. In this study, we explored whether miR-506 played a critical role in hepatocellular carcinoma (HCC).
Methods: Twenty HCC and adjacent normal liver tissue samples were collected. Human hepatoma cell lines HepG2 and H7402 were used for in vitro studies. The expression of miR-506 and transcriptional co-activator YAP was examined using qRT-PCR. Western blot analysis was used to measure the expression of YAP and its target genes. Luciferase reporter gene assay was used to identify YAP as a target gene of miR-506. MTT and EdU assays were carried out for functional analysis.
Results: The expression of miR-506 was significantly lower in HCC than in adjacent normal liver tissues. Bioinformatics analysis revealed that YAP mRNA might be one of the targets of miR-506, and miR-506 in HCC tissues was found to be negatively correlated with YAP (r=-0.605). In both HepG2 and H7402 cells, miR-506 dose-dependently down-regulated YAP and its target genes c-Myc and the connective tissue growth factor (CTGF). Luciferase reporter gene assays demonstrated that miR-506 targeted the wild type 3' UTR of YAP mRNA, but not a 3' UTR with a mutant seed site. Furthermore, miR-506 significantly inhibited the proliferation of HepG2 and H7402 cells, while anti-miR-506 enhanced the cell proliferation, which was blocked by YAP siRNA.
Conclusion: MiR-506 suppresses the proliferation of hepatoma cells by targeting YAP mRNA.
Keywords: microRNA; miR-506; hepatocellular carcinoma; cell proliferation; YAP; c-Myc; connective tissue growth factor
This work was supported by grants from the National Natural Science Foundation of China (No 81372186 and 81272218) and Support Program of National Science and Technology of China (No 2012BAI23B08).
* To whom correspondence should be addressed.
E-mail yelihong@nankai.edu.cn
Received 2014-03-24 Accepted 2014-05-18
Keywords:
1State Key Laboratory of Medicinal Chemical Biology, Department of Biochemistry, College of Life Sciences, Nankai University, Tianjin 300071, China; 2State Key Laboratory of Medicinal Chemical Biology, Department of Cancer Research, College of Life Sciences, Nankai University, Tianjin 300071, China
Aim: MiR-506 is a miRNA involved in carcinogenesis of several kinds of cancer. In this study, we explored whether miR-506 played a critical role in hepatocellular carcinoma (HCC).
Methods: Twenty HCC and adjacent normal liver tissue samples were collected. Human hepatoma cell lines HepG2 and H7402 were used for in vitro studies. The expression of miR-506 and transcriptional co-activator YAP was examined using qRT-PCR. Western blot analysis was used to measure the expression of YAP and its target genes. Luciferase reporter gene assay was used to identify YAP as a target gene of miR-506. MTT and EdU assays were carried out for functional analysis.
Results: The expression of miR-506 was significantly lower in HCC than in adjacent normal liver tissues. Bioinformatics analysis revealed that YAP mRNA might be one of the targets of miR-506, and miR-506 in HCC tissues was found to be negatively correlated with YAP (r=-0.605). In both HepG2 and H7402 cells, miR-506 dose-dependently down-regulated YAP and its target genes c-Myc and the connective tissue growth factor (CTGF). Luciferase reporter gene assays demonstrated that miR-506 targeted the wild type 3' UTR of YAP mRNA, but not a 3' UTR with a mutant seed site. Furthermore, miR-506 significantly inhibited the proliferation of HepG2 and H7402 cells, while anti-miR-506 enhanced the cell proliferation, which was blocked by YAP siRNA.
Conclusion: MiR-506 suppresses the proliferation of hepatoma cells by targeting YAP mRNA.
Keywords: microRNA; miR-506; hepatocellular carcinoma; cell proliferation; YAP; c-Myc; connective tissue growth factor
This work was supported by grants from the National Natural Science Foundation of China (No 81372186 and 81272218) and Support Program of National Science and Technology of China (No 2012BAI23B08).
* To whom correspondence should be addressed.
E-mail yelihong@nankai.edu.cn
Received 2014-03-24 Accepted 2014-05-18