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微小RNA-335-5p通过靶向八聚体转录因子4/蛋白激酶B信号通路抑制肝癌细胞Huh-7的增殖。

MiR-335-5p inhibits proliferation of Huh-7 liver cancer cells via targeting the Oct4/Akt pathway.

作者信息

Ji Y-Y, Song Y, Wang A-N

机构信息

Department of Infectious Diseases, Jiaozhou Central Hospital of Qingdao, Qingdao, China.

出版信息

Eur Rev Med Pharmacol Sci. 2021 Feb;25(4):1853-1860. doi: 10.26355/eurrev_202102_25080.

DOI:10.26355/eurrev_202102_25080
PMID:33660795
Abstract

OBJECTIVE

The aim of this study was to detect the expression of micro ribonucleic acid (miR)-335-5p in the liver tissues of patients with liver cancer, and to explore its effect on liver cancer and mechanism using Huh7 human liver cancer cells.

PATIENTS AND METHODS

Liver tissues were collected from patients with liver cancer. The expression of miR-335-5p in tissues was detected via quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Subsequently, Huh7 cells were transfected with miR-335-5p in vitro. After overexpressing miR-335-5p, changes in the expression of octamer-binding transcription factor 4 (Oct4) gene were observed via qRT-PCR. Furthermore, the proliferation of Huh7 cells and the protein expressions of protein kinase B (Akt) and phosphorylated Akt (p-Akt) were detected using cell counting kit (CCK)-8 assay and Western blotting (WB), respectively.

RESULTS

Compared with Control group, the expression of miR-335-5p increased significantly in the liver tissues of liver cancer patients (p<0.01). In comparison with those in negative group, the messenger RNA (mRNA) expression of Oct4 and the proliferation rate of Huh7 cells were both significantly inhibited in miR-335-5p group (p<0.01, p<0.05). After overexpression of miR-335-5p, the protein expression level of p-Akt decreased remarkably (p<0.01).

CONCLUSIONS

MiR-335-5p directly binds to the 3' untranslated region (3'UTR) of Oct4 mRNA to restrain the phosphorylation of Akt, thereby inhibiting Huh7 cell proliferation.

摘要

目的

本研究旨在检测肝癌患者肝组织中微小核糖核酸(miR)-335-5p的表达,并利用Huh7人肝癌细胞探讨其对肝癌的影响及机制。

患者与方法

收集肝癌患者的肝组织。通过定量逆转录-聚合酶链反应(qRT-PCR)检测组织中miR-335-5p的表达。随后,在体外将miR-335-5p转染至Huh7细胞。过表达miR-335-5p后,通过qRT-PCR观察八聚体结合转录因子4(Oct4)基因表达的变化。此外,分别使用细胞计数试剂盒(CCK)-8法和蛋白质免疫印迹法(WB)检测Huh7细胞的增殖以及蛋白激酶B(Akt)和磷酸化Akt(p-Akt)的蛋白表达。

结果

与对照组相比,肝癌患者肝组织中miR-335-5p的表达显著增加(p<0.01)。与阴性组相比,miR-335-5p组中Oct4的信使核糖核酸(mRNA)表达和Huh7细胞的增殖率均显著受到抑制(p<0.01,p<0.05)。miR-335-5p过表达后,p-Akt的蛋白表达水平显著降低(p<0.01)。

结论

MiR-335-5p直接与Oct4 mRNA的3'非翻译区(3'UTR)结合,抑制Akt的磷酸化,从而抑制Huh7细胞增殖。

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