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微小RNA-101通过靶向Fos抑制宫颈癌细胞从G1期到S期的转变。

miR-101 inhibits the G1-to-S phase transition of cervical cancer cells by targeting Fos.

作者信息

Liang Xiaolong, Liu Yi, Zeng Liqiong, Yu Chao, Hu Zhongwen, Zhou Qin, Yang Zhu

机构信息

*Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Chongqing Medical University; †Institute of Life Sciences; and ‡The M.O.E. Key Laboratory of Laboratory Medical Diagnostics, College of Laboratory Medicine, Chongqing Medical University, Chongqing, People's Republic of China.

出版信息

Int J Gynecol Cancer. 2014 Sep;24(7):1165-72. doi: 10.1097/IGC.0000000000000187.

Abstract

OBJECTIVES

The chief objective of this study was to identify the miRNAs targeting Fos, a well-recognized proto-oncogene that is commonly overexpressed in cervical cancer, and its biological significance on the cellular behaviors of HeLa, a cervical cancer cell.

MATERIALS AND METHODS

We initially analyzed the 3'untranslated region (3'UTR) of Fos and screened the potential miRNAs targeting Fos using 3 bioinformatical Web sites. Luciferase reporter assay, real-time polymerase chain reaction, and Western blotting were used to validate the binding of chosen miRNA (miR-101) on the 3'UTR of Fos and the downstream regulation on its mRNA and protein levels. Furthermore, flow cytometry along with the Fos rescue strategy was applied to analyze the modulation of cell cycle of HeLa cells by miR-101.

RESULTS

Among these predicted candidate miRNAs, miR-101 was the miRNAs preferred by all the 3 used Web sites. The results of luciferase reporter assay, real-time polymerase chain reaction, and Western blotting demonstrated that miR-101 directly targeted on the 3'UTR of Fos and down-regulated the expression of Fos at mRNA and protein levels. Furthermore, cell cycle analysis showed that miR-101 arrests G1-to-S phase transition of HeLa cells, at least partially by targeting Fos.

CONCLUSIONS

We concluded that by targeting the proto-oncogene Fos, miR-101 is involved in G1-to-S phase transition in cervical cancer cells in vitro and might provide a new approach for the pharmacological interference node in cervical cancer treatment.

摘要

目的

本研究的主要目的是鉴定靶向Fos的微小RNA(miRNA),Fos是一种公认的原癌基因,在宫颈癌中通常过度表达,以及其对宫颈癌细胞HeLa细胞行为的生物学意义。

材料与方法

我们首先分析了Fos的3'非翻译区(3'UTR),并使用3个生物信息学网站筛选靶向Fos的潜在miRNA。荧光素酶报告基因检测、实时聚合酶链反应和蛋白质免疫印迹法用于验证所选miRNA(miR-101)与Fos的3'UTR的结合及其对其mRNA和蛋白质水平的下游调控。此外,采用流式细胞术结合Fos拯救策略分析miR-101对HeLa细胞周期的调控作用。

结果

在这些预测的候选miRNA中,miR-101是所有3个使用的网站都首选的miRNA。荧光素酶报告基因检测、实时聚合酶链反应和蛋白质免疫印迹法的结果表明,miR-101直接靶向Fos的3'UTR,并在mRNA和蛋白质水平下调Fos的表达。此外,细胞周期分析表明,miR-101至少部分通过靶向Fos阻止HeLa细胞从G1期向S期转变。

结论

我们得出结论,通过靶向原癌基因Fos,miR-101参与体外宫颈癌细胞从G1期向S期的转变,并可能为宫颈癌治疗中的药理干预节点提供一种新方法。

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