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多西他赛通过上调微小RNA-7的表达来抑制非小细胞肺癌细胞的增殖。

Docetaxel inhibits the proliferation of non-small-cell lung cancer cells via upregulation of microRNA-7 expression.

作者信息

He Xigan, Li Chunxia, Wu Xiaoyan, Yang Guotao

机构信息

Department of Thoracic Surgery, Qilu Hospital of Shandong University Jinan, China.

出版信息

Int J Clin Exp Pathol. 2015 Aug 1;8(8):9072-80. eCollection 2015.

Abstract

Lung cancer is the leading cause of cancer-related deaths worldwide and about 85% of these are non-small cell lung cancer (NSCLC). Several new chemotherapeutic agents have recently shown encouraging activity in NSCLC, especially docetaxel. MiRNAs (MicroRNAs) are closely related to cancer development. We studied miRNAs in NSCLC cell lines to identify those that can regulate and predict the effectiveness of docetaxel on NSCLC. CCK8, Annexin and V-FITC assays were carried out to evaluate the inhibitory effect of docetaxel on NSCLC cell lines A549 and H460, and qRT-PCR was used to detect and compare six miRNAs expression levels in the two cells with docetaxel or not. Knockdown of miR-7 by RNA interference and overexpression of miR-7 were taken to evaluate the effect of miR-7 on docetaxel effectiveness. Western blotting was used to evaluate the effect of miR-7 on Bcl2 in A549 and H460 cells. Docetaxel induced non-small cell lung cancer cell apoptosis and suppressed cell proliferation in vitro. MiR-7 expression levels were increased by docetaxel in the two cell lines. MiR-7 overexpression improved anti-proliferative and pro-apoptotic effects of docetaxel on the NSCLC cells and that miR-7 down-regulation decreased those effects. Moreover, subsequent experiments showed that BCL-2 was downregulated by miR-7 at both transcriptional and translational levels. This study further extends the biological role of miR-7 in NSCLC A549 and H460 cells and identifies BCL-2 as a novel target possibly involved in miR-7-mediated growth suppression and apoptosis induction of NSCLC cells.

摘要

肺癌是全球癌症相关死亡的主要原因,其中约85%为非小细胞肺癌(NSCLC)。最近,几种新型化疗药物在NSCLC中显示出令人鼓舞的活性,尤其是多西他赛。微小RNA(miRNA)与癌症发展密切相关。我们研究了NSCLC细胞系中的miRNA,以确定那些能够调节并预测多西他赛对NSCLC疗效的miRNA。进行CCK8、膜联蛋白和V-FITC检测以评估多西他赛对NSCLC细胞系A549和H460的抑制作用,并使用qRT-PCR检测和比较两种细胞在有无多西他赛情况下六种miRNA的表达水平。通过RNA干扰敲低miR-7以及过表达miR-7来评估miR-7对多西他赛疗效的影响。使用蛋白质免疫印迹法评估miR-7对A549和H460细胞中Bcl2的影响。多西他赛在体外可诱导非小细胞肺癌细胞凋亡并抑制细胞增殖。在这两种细胞系中,多西他赛均可使miR-7表达水平升高。miR-7过表达可增强多西他赛对NSCLC细胞的抗增殖和促凋亡作用,而miR-7下调则会减弱这些作用。此外,后续实验表明,miR-7在转录和翻译水平均下调BCL-2。本研究进一步扩展了miR-7在NSCLC A549和H460细胞中的生物学作用,并确定BCL-2是一个可能参与miR-7介导的NSCLC细胞生长抑制和凋亡诱导的新靶点。

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