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Silencing livin gene by siRNA leads to apoptosis induction, cell cycle arrest, and proliferation inhibition in malignant melanoma LiBr cells
作者姓名:Wang H  Tan SS  Wang XY  Liu DH  Yu CS  Bai ZL  He DL  Zhao J
作者单位:[1]Department of Dermatology, The Second Affliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China [2]Institute of Urology, Key Laboratory of Environment and Genes Related to Diseases of Ministry of Education, Xi'an Jiaotong University, Xi'an 710061,China [3]Department of Urology, The Medical School Hospital of Qingdao University, Qingdao 266003, China
摘    要:Aim: The aim of the present study was to investigate the effects of silencing the livin gene by small interfering RNA (siRNA) on the expression of livin and the effects on apoptosis, cell cycle, and proliferation in human malignant melanoma LiBr cells. Methods: Three chemically-synthetic siRNA duplexes targeting livin were transiently transfected into the LiBr cells, and the effects on livin expression were detected both at the mRNA level by real-time RT-PCR and at the protein level by Western blotting. Apoptosis was evaluated by terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling assay, flow cytometric analysis, and the expression of procaspase-3 and activated caspase-3 analysis by Western blotting. Cell cycle was analyzed by flow cytometry. Cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. Results: One of the 3 designed siRNA could effectively knock down the livin expression both at the mRNA and protein levels in dose- and time-dependent manners; 100 nmol/L with maximum downregulation on mRNA at 48 h, and on the protein at 72 h after transfection. Silencing livin could significantly induce apoptosis, arrest cell cycle at the Go/G1 phase, and inhibit proliferation in LiBr cells. Meanwhile, caspase-3 was activated. Conclusion: The livin gene could serve as a potential molecular target for gene therapy by siRNA for malignant melanoma.

关 键 词:黑色素瘤  细胞凋亡  细胞周期  增殖
收稿时间:2007-06-15
修稿时间:2007-08-17

Silencing livin gene by siRNA leads to apoptosis induction, cell cycle arrest, and proliferation inhibition in malignant melanoma LiBr cells
Wang H,Tan SS,Wang XY,Liu DH,Yu CS,Bai ZL,He DL,Zhao J.Silencing livin gene by siRNA leads to apoptosis induction, cell cycle arrest, and proliferation inhibition in malignant melanoma LiBr cells[J].Acta Pharmacologica Sinica,2007,28(12):1968-1974.
Authors:Wang Hao  Tan Sheng-shun  Wang Xin-yang  Liu Dong-hua  Yu Chun-shui  Bai Zhuan-li  He Da-lin  Zhao Jun
Institution:Department of Dermatology, The Second Affliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China.
Abstract:AIM: The aim of the present study was to investigate the effects of silencing the livin gene by small interfering RNA (siRNA) on the expression of livin and the effects on apoptosis, cell cycle, and proliferation in human malignant melanoma LiBr cells. METHODS: Three chemically-synthetic siRNA duplexes targeting livin were transiently transfected into the LiBr cells, and the effects on livin expression were detected both at the mRNA level by real-time RT-PCR and at the protein level by Western blotting. Apoptosis was evaluated by terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling assay, flow cytometric analysis, and the expression of procaspase-3 and activated caspase-3 analysis by Western blotting. Cell cycle was analyzed by flow cytometry. Cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. RESULTS: One of the 3 designed siRNA could effectively knock down the livin expression both at the mRNA and protein levels in dose- and time-dependent manners; 100 nmol/L with maximum downregulation on mRNA at 48 h, and on the protein at 72 h after transfection. Silencing livin could significantly induce apoptosis, arrest cell cycle at the G0/G1 phase, and inhibit proliferation in LiBr cells. Meanwhile, caspase-3 was activated. CONCLUSION: The livin gene could serve as a potential molecular target for gene therapy by siRNA for malignant melanoma.
Keywords:livin  malignant melanoma  siRNA  apoptosis  cell cycle  proliferation
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