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[微小RNA-34a通过靶向人膀胱癌细胞中的CD44调控细胞周期]

[MicroRNA-34a regulates cell cycle by targeting CD44 in human bladder carcinoma cells].

作者信息

Yu Gan, Xu Kai, Xu Shian, Zhang Xiaolan, Huang Qianhua, Lang Bin

机构信息

Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.E-mail:

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2015 Jul;35(7):935-40.

Abstract

OBJECTIVE

To investigate the role of microRNA-34a (miR-34a) in regulating the cell cycles of bladder cancer cell line J82 and explore the underlying mechanism.

METHODS

J82 cells were transfected with a miR-34a mimic or an inhibitor to induce miR-34a overexpression or silencing. The RNA level of miR-34a in the transfected cells was detected by real-time PCR, and CD44 expressions at the mRNA and protein levels were detected by real-time PCR and Western blotting. Luciferase reporter assay was used to detect the activation of 3'UTR of CD44, and flow cytometry was performed to analyze the cell cycle changes.

RESULTS

The expression level of miR-34a was significantly increased and CD44 expression significantly lowered in cells transfected with miR-34a mimic; miR-34a inhibitor transfection caused reverse effects on miR-34a and CD44 expressions. MiR-34a mimics downregulated while miR-34a inhibitor enhanced the activation of 3'UTR of CD44 with corresponding changes in the expressions of some cell cycle-related proteins. MiR-34a mimics and miR-34a inhibitor induced opposite changes in J82 cell cycle, which were partly reversed by CD44.

CONCLUSION

MiRNA-34a regulates cell cycles by targeting CD44 in human bladder carcinoma cell line J82.

摘要

目的

探讨微小RNA-34a(miR-34a)在调控膀胱癌细胞系J82细胞周期中的作用,并探究其潜在机制。

方法

用miR-34a模拟物或抑制剂转染J82细胞,以诱导miR-34a过表达或沉默。通过实时PCR检测转染细胞中miR-34a的RNA水平,通过实时PCR和蛋白质印迹法检测CD44在mRNA和蛋白质水平的表达。采用荧光素酶报告基因检测法检测CD44 3'UTR的激活情况,并通过流式细胞术分析细胞周期变化。

结果

用miR-34a模拟物转染的细胞中,miR-34a表达水平显著升高,CD44表达显著降低;转染miR-34a抑制剂则对miR-34a和CD44的表达产生相反的影响。miR-34a模拟物下调而miR-34a抑制剂增强CD44 3'UTR的激活,同时一些细胞周期相关蛋白的表达也发生相应变化。miR-34a模拟物和miR-34a抑制剂诱导J82细胞周期发生相反变化,而CD44可部分逆转这些变化。

结论

在人膀胱癌细胞系J82中,miRNA-34a通过靶向CD44调控细胞周期。

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