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敲低EIF3D通过G2/M期阻滞抑制人黑色素瘤细胞的增殖。

Knockdown of EIF3D suppresses proliferation of human melanoma cells through G2/M phase arrest.

作者信息

Li Hui, Zhou Fusheng, Wang Hongyan, Lin Da, Chen Gang, Zuo Xianbo, Sun Liangdan, Zhang Xuejun, Yang Sen

机构信息

Department of Dermatology, Institute of Dermatology, The First Affiliated Hospital, Anhui Medical University, Hefei, Anhui, People's Republic of China.

出版信息

Biotechnol Appl Biochem. 2015 Sep-Oct;62(5):615-20. doi: 10.1002/bab.1305. Epub 2015 Jan 12.

Abstract

Skin cancer is the most common malignancy with increasing incidence rates worldwide. The advanced form of skin cancer, melanoma, is resistant to conventional treatment methods, which motivated researchers to identify an alternative effective therapeutic approach. This study was designed to identify the effects of small interfering RNA (si-RNA) mediated silencing of eukaryotic translation initiation factor 3, subunit D (EIF3D) against melanoma cell survival. Briefly, a lentivirus-mediated RNA interference system was employed to knock down EIF3D expression in A375 and A431 melanoma cells. The cell proliferation was analyzed by methylthiazoletetrazolium (MTT) and colony formation assays. The cell cycle progression was investigated using flow cytometry. Results revealed that si-RNA-mediated knockdown of EIF3D significantly reduced the gene and protein expression levels of EIF3D in melanoma cells. Furthermore, knockdown of EIF3D led to a significant reduction in cell proliferation due to G2 /M phase cell cycle arrest. Apparently, the study demonstrated the critical involvement of EIF3D in the survival and progression of melanoma cells and depletion of EIF3D could be developed as a possible therapeutic option in the gene-targeted treatment of melanoma.

摘要

皮肤癌是全球发病率不断上升的最常见恶性肿瘤。皮肤癌的晚期形式黑色素瘤对传统治疗方法具有抗性,这促使研究人员寻找另一种有效的治疗方法。本研究旨在确定小干扰RNA(si-RNA)介导的真核翻译起始因子3亚基D(EIF3D)沉默对黑色素瘤细胞存活的影响。简而言之,采用慢病毒介导的RNA干扰系统来敲低A375和A431黑色素瘤细胞中EIF3D的表达。通过甲基噻唑基四氮唑(MTT)和集落形成试验分析细胞增殖。使用流式细胞术研究细胞周期进程。结果显示,si-RNA介导的EIF3D敲低显著降低了黑色素瘤细胞中EIF3D的基因和蛋白表达水平。此外,EIF3D的敲低由于G2/M期细胞周期阻滞导致细胞增殖显著减少。显然,该研究证明了EIF3D在黑色素瘤细胞存活和进展中的关键作用,EIF3D的缺失可作为黑色素瘤基因靶向治疗的一种可能的治疗选择。

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