王 建,何志巍.靶向hTERT的小干扰RNA诱导前列腺癌PC3细胞凋亡的机制[J].中国肿瘤,2012,21(3):211-214. |
靶向hTERT的小干扰RNA诱导前列腺癌PC3细胞凋亡的机制 |
Mechanism of Induction of Apoptosis by siRNA Targeting hTERT in Prostatic Cancer PC3 Cells |
投稿时间:2011-08-26 |
DOI: |
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中文关键词: 端粒酶逆转录酶 RNA干扰 凋亡 前列腺癌 |
英文关键词:hTERT RNAi apoptosis prostatic cancer |
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中文摘要: |
摘 要:[目的] 研究靶向人端粒酶逆转录酶(hTERT)的siRNA诱导前列腺癌PC3细胞凋亡的分子机制。[方法] 化学合成的靶向hTERT的siRNA以脂质体法转染PC3细胞;应用寡核苷酸芯片技术分析细胞凋亡相关基因的转录谱变化;应用Western blot检测细胞中hTERT、肿瘤坏死因子相关凋亡诱导配体TRAIL、凋亡抑制基因Bcl-2和胞浆Cyt c的蛋白水平变化;流式细胞术分析细胞凋亡率变化;比色法测定Caspase-3和Caspase-8的相对活性改变。[结果] 靶向hTERT的siRNA能有效抑制hTERT基因表达;转染hTERT-siRNA 48h后可引起TRAIL的表达上调,Bcl-2表达下调,Cyt c释放增加,Caspase-3活性增强,细胞凋亡率升高。[结论] 靶向hTERT的小干扰RNA通过活化线粒体信号传导途径诱导PC3细胞凋亡。 |
英文摘要: |
Abstract: [Purpose]To study the molecular mechanism of induction of apoptosis by siRNA targeting hTERT in prostatic cancer PC3 Cells. [Methods] PC3 cells were transfected with chemical synthesized siRNA-hTERT by liposome method. The microarray assay was adopted to explore the transcriptional profiling of apoptosis associated genes. The protein levels of hTERT, TRAIL, Bcl-2 and cytosolic Cyt c were detected by Western blot assay. The apoptosis rate was determined by flow cytometry. The relative activity of Caspase-3 and Caspase-8 was measured by colorimetric assay. [Results] The siRNA-hTERT suppressed the expression of hTERT gene significantly. Forty-eight hours after transfection, the expression level of TRAIL was increased, the expression of Bcl-2 was down regulation, the releasing of cytosolic Cytc enhanced, the activation of Caspase-3 increased, and the apoptosis rate increased.[Conclusion] siRNA-hTERT induces apoptosis of PC3 cells via activating mitochondrial signal transduction pathway. |
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