Niu X, Wei N, Peng L, Li X, Zhang X, Wang C
Zhuhai Campus of Zunyi Medical University, Department of Biochemistry and Molecular Biology, Zhuhai, Guangdong, China.
Fifth Affiliated (Zhuhai) Hospital of Zunyi Medical University, Department of Oncology, Zhuhai, Guangdong, China.
Malays J Pathol. 2022 Apr;44(1):39-52.
The objective of this research is to determine the role of miR-34a-5p in the occurrence and development of HCC by targeting VEGFA.
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.
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.
These results suggest that miR-34a-5p could inhibit the occurrence and development of HCC by targeting VEGFA.
本研究旨在通过靶向血管内皮生长因子A（VEGFA）来确定miR - 34a - 5p在肝癌（HCC）发生发展中的作用。
采用qRT - PCR检测miR - 34a - 5p在肝癌细胞系和肿瘤组织中的表达；通过Transwell侵袭实验检测miR - 34a - 5p对肝癌细胞（SMMC7721和MHCC97H）侵袭能力的影响；利用TargetScan预测VEGFA为miR - 34a - 5p的潜在靶基因，并通过双荧光素酶报告基因实验、qRT - PCR和蛋白质印迹法进行验证。采用qRT - PCR检测肝癌细胞系和肿瘤组织中VEGFA的表达；通过CCK8、Transwell实验、划痕实验和流式细胞术检测miR - 34a - 5p和VEGFA对不同侵袭能力肝癌细胞的增殖、侵袭、迁移及S期细胞的调控和影响。通过构建肝癌裸鼠异种移植模型检测miR - 34a - 5p对肿瘤生长的影响。
发现miR - 34a - 5p在肝癌细胞和肿瘤组织中的表达显著降低。上调miR - 34a - 5p表达可降低肝癌细胞的侵袭能力。miR - 34a - 5p可通过与VEGFA的3'-UTR结合抑制VEGFA的mRNA和蛋白表达水平。VEGFA在肝癌细胞和肿瘤组织中高表达。miR - 34a - 5p抑制肝癌细胞的增殖、侵袭、迁移及S期阻滞，但这种抑制作用可被VEGFA中和；在肝癌裸鼠异种移植模型中，miR - 34a - 5p对肝癌细胞增殖和肿瘤生长发挥抑制作用。
这些结果表明，miR - 34a - 5p可通过靶向VEGFA抑制肝癌的发生发展。
