微小RNA-25通过细胞周期蛋白E2调节小细胞肺癌细胞的发育和细胞周期。
MicroRNA-25 regulates small cell lung cancer cell development and cell cycle through cyclin E2.
作者信息
Zhao Zhengyuan, Liu Juntao, Wang Changlei, Wang Yi, Jiang Youguo, Guo Min
机构信息
Department of General Thoracic Surgery, Affiliated Hospital of Logistics University of Chinese People's Armed Police Forces Tianjin 300162, China.
出版信息
Int J Clin Exp Pathol. 2014 Oct 15;7(11):7726-34. eCollection 2014.
PURPOSE
We intended to examine the underlying mechanism of microRNA-25 (miR-25) in regulating small cell lung cancer (SCLC).
METHODS
The miR-25 expression was measured by quantitative RT-PCR (qRT-PCR) in 5 SCLC cell lines and 9 human SCLC tissues. In SCLC cell line H510A cells, endogenous miR-25 was downregulated by stable transfection of antisense oligonucleotide of miR-25 (miR-25-as). Then the effects of miR-25 downregulation on SCLC growth, invasion and chemoresistance were assessed by MTT, migration and cisplatin assays, respectively. Furthermore, the effects of miR-25 downregulation on cancer cell cycle arrest, production of cell cycle proteins cyclin E2 and CDK2 were examined by cell cycle assay, western blot and luciferase assays, respectively. Finally, cyclin E2 was over-expressed in H510A cells to investigate its effect on miR-25 mediated SCLC regulation.
RESULTS
In both SCLC cells and human SCLC tumor tissues, miR-25 was overexpressed. Down-regulation of miR-25 in H510A cells significantly reduced cancer cell growth, invasive capability and resistance to cisplatin. Also, it induced G1 cell cycle arrest and downregulated cell cycle related proteins cyclin E2 and CDK2. Luciferase assay demonstrated cyclin E2 was directly targeted by miR-25. Overexpression of cyclin E2 in H510A cells reversed the cell cycle arrest and restored invasive capability impaired by miR-25 downregulation.
CONCLUSIONS
Our study shows miR-25 is overexpressed in SCLC and acting as oncogenic regulator by regulating cyclin E2.
目的
我们旨在研究微小RNA - 25（miR - 25）调控小细胞肺癌（SCLC）的潜在机制。
方法
采用定量逆转录聚合酶链反应（qRT - PCR）检测5种SCLC细胞系和9例人SCLC组织中miR - 25的表达。在SCLC细胞系H510A细胞中，通过稳定转染miR - 25反义寡核苷酸（miR - 25 - as）下调内源性miR - 25。然后分别通过MTT法、迁移实验和顺铂实验评估miR - 25下调对SCLC生长、侵袭和化疗耐药性的影响。此外，分别通过细胞周期实验、蛋白质免疫印迹法和荧光素酶实验检测miR - 25下调对癌细胞周期阻滞、细胞周期蛋白细胞周期蛋白E2（cyclin E2）和细胞周期蛋白依赖性激酶2（CDK2）产生的影响。最后，在H510A细胞中过表达cyclin E2，以研究其对miR - 25介导的SCLC调控的影响。
结果
在SCLC细胞和人SCLC肿瘤组织中，miR - 25均过表达。H510A细胞中miR - 25的下调显著降低了癌细胞的生长、侵袭能力和顺铂耐药性。此外，它诱导G1期细胞周期阻滞，并下调细胞周期相关蛋白cyclin E2和CDK2。荧光素酶实验表明cyclin E2是miR - 25的直接靶标。H510A细胞中cyclin E2的过表达逆转了细胞周期阻滞，并恢复了因miR - 25下调而受损的侵袭能力。
结论
我们的研究表明，miR - 25在SCLC中过表达，并通过调控cyclin E2发挥致癌调节作用。
Zotero 插件