Zhang J, Zhao M, Xue Z-Q, Liu Y, Wang Y-X
Department of Thoracic Surgery, Chinese PLA General Hospital, Haiding, Beijing, China.
Eur Rev Med Pharmacol Sci. 2016 Nov;20(21):4494-4499.
This study is aimed to investigate the cellular role of miR-377 and demonstrate that miR-377 negatively regulated cyclin-dependent kinase 6(CDK6) in human non-small cell lung cancer (NSCLC) cells.
qRT-PCR was performed to identify the miR-377 expression level in 45 paired NSCLC and adjacent normal lung tissues. Cell proliferation was measured by MTT. Apoptosis was detected by flow cytometric analysis. Luciferase reporter assays were employed to validate regulation of a putative target of miR-377. The effect of miR-377 on endogenous levels of this target was subsequently confirmed via Western blot.
We found that the expression level of miR-377 was significantly reduced in NSCLC tissues and cell lines. On the contrary, CDK6 expression level was up-regulated in NSCLC tissues and cell lines. Based on Luciferase reporter assays, we confirmed that CDK6 was a direct target gene of miR-377. In vitro studies demonstrated that miR-377 overexpression reduced NSCLC cell proliferation and promoted apoptosis.
Our discovery suggested that miR-377 might be used as a therapeutic reagent for the treatment of NSCLC in the future.
本研究旨在探讨miR-377的细胞作用,并证明miR-377在人非小细胞肺癌(NSCLC)细胞中对细胞周期蛋白依赖性激酶6(CDK6)具有负调控作用。
采用qRT-PCR检测45对NSCLC组织及癌旁正常肺组织中miR-377的表达水平。通过MTT法检测细胞增殖情况。采用流式细胞术分析检测细胞凋亡情况。利用荧光素酶报告基因检测法验证miR-377假定靶标的调控作用。随后通过蛋白质印迹法证实miR-377对该靶标内源性水平的影响。
我们发现NSCLC组织和细胞系中miR-377的表达水平显著降低。相反,NSCLC组织和细胞系中CDK6的表达水平上调。基于荧光素酶报告基因检测法,我们证实CDK6是miR-377的直接靶基因。体外研究表明,miR-377过表达可降低NSCLC细胞增殖并促进细胞凋亡。
我们的发现表明,miR-377未来可能用作治疗NSCLC的治疗试剂。