Huang Junchao, Sun Chengchao, Wang Suqing, He Qiqiang, Li Dejia
Hubei Province Key Laboratory on Cardiovascular, Cerebrovascular, and Metabolic Disorders, School of Nuclear Technology and Chemistry & Biology, Hubei University of Science and Technology, Xianning, 437100, China.
Mol Biosyst. 2015 Jul;11(7):2051-9. doi: 10.1039/c4mb00752b.
Lung cancer is one of the most common and serious types of cancer. Till now, the treatment of lung cancer has been unsatisfactory, which is associated with poor prognosis and high mortality. Therefore, there is an urgent requirement to investigate the molecular mechanisms underlying lung tumorigenesis. To study the potential function of miR-10b involved in the regulation of lung tumors, we monitored NSCLC cell behaviour including proliferation, apoptosis and cell cycle using CCK-8 and flow cytometry analysis. Real-time PCR was used to detect the expression levels of miR-10b in 75 NSCLC patients' tissues and Western blot was also used to analyze the expression level of genes correlated with apoptosis in NSCLC cells. miR-10b expression levels were higher in NSCLC tissues compared with an adjacent normal tissue control. Silencing of miR-10b inhibited cancer cell progress by arresting cell cycle progression in the G0/G1 phase and promoted apoptosis in NSCLC cells. Western blot analysis of miR-10b-silenced cells revealed up-regulation of apoptosis-inducing members Fas, FasL, Bax and caspase 3, and down-regulation of apoptosis-inhibiting factors Bcl-2 and PCNA. And, a significant inverse correlation between the level of miR-10b and klotho was observed, which has been demonstrated to be a novel tumor suppressor gene. A further in vivo tumor formation study in nude mice indicated that inhibition of miR-10b in lung cancer cells delayed the progress of tumor formation. These findings indicated that miR-10b might serve as a useful potential target for treatment of NSCLC.
肺癌是最常见且最严重的癌症类型之一。到目前为止,肺癌的治疗效果并不理想,其预后较差且死亡率高。因此,迫切需要研究肺癌发生的分子机制。为了研究参与调控肺肿瘤的miR-10b的潜在功能,我们使用CCK-8和流式细胞术分析监测了非小细胞肺癌(NSCLC)细胞的行为,包括增殖、凋亡和细胞周期。采用实时定量PCR检测75例NSCLC患者组织中miR-10b的表达水平,并用蛋白质免疫印迹法分析NSCLC细胞中与凋亡相关基因的表达水平。与相邻正常组织对照相比,NSCLC组织中miR-10b表达水平更高。沉默miR-10b可通过使细胞周期停滞在G0/G1期来抑制癌细胞进展,并促进NSCLC细胞凋亡。对miR-10b沉默细胞进行蛋白质免疫印迹分析显示,凋亡诱导成员Fas、FasL、Bax和半胱天冬酶3上调,凋亡抑制因子Bcl-2和增殖细胞核抗原(PCNA)下调。此外,观察到miR-10b水平与klotho之间存在显著负相关,klotho已被证明是一种新的肿瘤抑制基因。在裸鼠中进行的进一步体内肿瘤形成研究表明,抑制肺癌细胞中的miR-10b可延缓肿瘤形成进程。这些发现表明,miR-10b可能是治疗NSCLC的一个有用的潜在靶点。