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肌球蛋白VI的下调可降低人结直肠癌中的细胞生长并增加细胞凋亡。

Downregulation of myosin VI reduced cell growth and increased apoptosis in human colorectal cancer.

作者信息

You Weiqiang, Tan Gewen, Sheng Nengquan, Gong Jianfeng, Yan Jun, Chen Di, Zhang Huizhen, Wang Zhigang

机构信息

Department of General Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai 200233, China.

National Key Laboratory of Science and Technology on Nano/Micro Fabrication Technology, Research Institute Micro/Nano Science and Technology, Shanghai Jiao Tong University, Shanghai 200240, China.

出版信息

Acta Biochim Biophys Sin (Shanghai). 2016 May;48(5):430-6. doi: 10.1093/abbs/gmw020. Epub 2016 Apr 3.

Abstract

Colorectal cancer (CRC) is the third most commonly diagnosed cancer worldwide, with the mortality increasing steadily over the last decade. Myosin VI (MYO6) expression is found to be elevated in some types of human carcinoma cell types, suggesting that it may be a sensitive biomarker for the diagnosis and follow-up. In this study, we first used the Oncomine database to explore the expression of MYO6 in CRC tissues, and then constructed a plasmid of RNA interference targeting MYO6 gene. After transfection of lentivirus targeting MYO6 into SW1116 cells, cell viability and proliferation were measured with 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and colony formation assay. Cell cycle distribution was assayed by flow cytometry and apoptosis was evaluated by Annexin V. MYO6 expression was detected by quantitative real-time polymerase chain reaction and western blot analysis. It was found that MYO6 mRNA was upregulated in CRC tissues using data mining of public Oncomine microarray datasets. Depletion of MYO6 significantly inhibited cell proliferation and colony formation. In addition, knockdown of MYO6 slightly arrested cell cycle in G0/G1 phase, but remarkably increased the proportion of the sub-G1 phase of cell with the increase of apoptotic cells. These results suggest that MYO6 may promote cell growth and may be used as a potential target for anticancer therapy of CRC.

摘要

结直肠癌(CRC)是全球第三大常见诊断癌症,在过去十年中死亡率稳步上升。研究发现肌球蛋白VI(MYO6)在某些类型的人类癌细胞中表达升高,这表明它可能是诊断和随访的敏感生物标志物。在本研究中,我们首先使用Oncomine数据库探索MYO6在CRC组织中的表达,然后构建靶向MYO6基因的RNA干扰质粒。将靶向MYO6的慢病毒转染到SW1116细胞后,用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐和集落形成试验测量细胞活力和增殖。通过流式细胞术检测细胞周期分布,通过膜联蛋白V评估细胞凋亡。通过定量实时聚合酶链反应和蛋白质印迹分析检测MYO6表达。利用公共Oncomine微阵列数据集的数据挖掘发现,MYO6 mRNA在CRC组织中上调。MYO6的缺失显著抑制细胞增殖和集落形成。此外,敲低MYO6会使细胞周期在G0/G1期略有停滞,但随着凋亡细胞的增加,显著增加了亚G1期细胞的比例。这些结果表明,MYO6可能促进细胞生长,并可能作为CRC抗癌治疗的潜在靶点。

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