小干扰RNA敲低生存素表达可诱导人乳腺癌细胞系MCF-7凋亡。
Knockdown of survivin expression by small interfering RNA induces apoptosis in human breast carcinoma cell line MCF-7.
作者信息
Li Ji-Xia, Zhou Ke-Yuan, Liang Tong, Zhang Yue-Fei
机构信息
Institute of Biochemistry and Molecular Biology, Guangdong Medical College, Zhanjiang, Guangdong, 524023, P.R.China.
出版信息
Ai Zheng. 2005 Mar;24(3):268-72.
BACKGROUND & OBJECTIVE: survivin, a member of inhibitor of apoptosis protein (IAP) gene family, expresses in various human cancer tissues, and may facilitate tumor cell evasion from apoptosis, and promote aberrant mitotic progression. This study was to investigate cell proliferation and apoptosis status of human breast carcinoma cell line MCF-7 after knockdown of survivin.
METHODS
Small interfering RNA was transfected into MCF-7 cells to inhibit expression of survivin. mRNA and protein levels of survivin were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Proliferation inhibition rate of MCF-7 cells was analyzed by MTT assay. Cell apoptosis was assayed by flow cytometry (FCM).
RESULTS
The expression of survivin in siRNA-transfected group decreased by 64% in comparison to untransfected group. After treatment of different concentrations of siRNA, proliferation inhibition rate and apoptosis rate of MCF-7 cells were increased. The highest proliferation inhibition rate was 60.9%, and the highest apoptosis rate was 29.0% after treatment of 200 nmol/L of siRNA.
CONCLUSION
survivin siRNA might be a useful therapeutic agent for the treatment of breast carcinoma.
背景与目的
生存素是凋亡抑制蛋白（IAP）基因家族的成员之一，在多种人类癌症组织中表达，可能促使肿瘤细胞逃避凋亡，并促进异常的有丝分裂进程。本研究旨在探讨生存素基因敲低后人乳腺癌细胞系MCF-7的细胞增殖和凋亡状态。
方法
将小干扰RNA转染至MCF-7细胞以抑制生存素的表达。通过逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测生存素的mRNA和蛋白水平。采用MTT法分析MCF-7细胞的增殖抑制率。通过流式细胞术（FCM）检测细胞凋亡情况。
结果
与未转染组相比，siRNA转染组生存素的表达下降了64%。用不同浓度的siRNA处理后，MCF-7细胞的增殖抑制率和凋亡率均升高。用200 nmol/L的siRNA处理后，最高增殖抑制率为60.9%，最高凋亡率为29.0%。
结论
生存素siRNA可能是治疗乳腺癌的一种有效治疗药物。
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