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Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
miR-34a-5p plays an inhibitory role in hepatocellular carcinoma by regulating target gene VEGFA
1Zhuhai Campus of Zunyi Medical University, Department of Biochemistry and Molecular Biology, Zhuhai, Guangdong, China. niuxianli2019@163.com.
Objective:
The objective of this research is to determine the role of miR-34a-5p in the occurrence and development of HCC by targeting VEGFA.
Methods:
The expression of miR-34a-5p in HCC cell lines and tumour tissue was detected by qRT-PCR; the effect of miR-34a-5p on the invasive ability of HCC cells (SMMC7721 and MHCC97H) were detected by Transwell invasion assay; VEGFA is predicted as a potential target gene of miR-34a-5p by TargetScan, and validated with dual-luciferase reporter gene assay, qRT-PCR and western blot. VEGFA expression in HCC cell lines and tumour tissue was detected using qRT-PCR; the regulation and influence of miR-34a-5p and VEGFA on the proliferation, invasion, migration and the S-phase cell of HCC cells with different invasive abilities were detected by CCK8, Transwell assay, wound healing assay, and flow cytometry. The effect of miR-34a-5p on the growth of tumour was detected by constructing a xenograft model of nude mice with HCC.
Results:
It was found that the expression of miR-34a-5p in HCC cells and tumour tissue was significantly decreased. Up-regulating miR-34a-5p expression could reduce the invasion ability of HCC cells. MiR-34a-5p could inhibit the mRNA and protein expression level of VEGFA via combining with the 3'-UTR of VEGFA. VEGFA was highly expressed in HCC cells and tumour tissues. The miR-34a-5p inhibited the proliferation, invasion, migration and S-phase arrest of HCC cells, but this inhibition effect could be neutralised by VEGFA; miR-34a-5p exerted the inhibitory effect on HCC cell proliferation and tumour growth in the HCC xenograft model of nude mice.
Conclusion:
These results suggest that miR-34a-5p could inhibit the occurrence and development of HCC by targeting VEGFA.
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