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微小RNA-34a-5p通过调控靶基因血管内皮生长因子A(VEGFA)在肝细胞癌中发挥抑制作用。

miR-34a-5p plays an inhibitory role in hepatocellular carcinoma by regulating target gene VEGFA.

作者信息

Niu X, Wei N, Peng L, Li X, Zhang X, Wang C

机构信息

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.

出版信息

Malays J Pathol. 2022 Apr;44(1):39-52.

Abstract

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.

摘要

目的

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。

方法

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。

结果

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。

结论

这些结果表明,miR - 34a - 5p可通过靶向VEGFA抑制肝癌的发生发展。

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