引用本文:黄 彭,苟 欣,何卫阳,罗 勇.多西紫杉醇诱导前列腺癌PC-3细胞自噬的研究[J].重庆医科大学学报,2014,38(6):797~800
多西紫杉醇诱导前列腺癌PC-3细胞自噬的研究
Research of autophagy induced by docetaxel in prostate cancer cell line PC-3
DOI:
中文关键词:  前列腺癌  多西紫杉醇  自噬  PC-3
英文关键词:prostate cancer  docetaxel  autophagy  PC-3
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作者单位
黄 彭,苟 欣,何卫阳,罗 勇 重庆医科大学附属第一医院泌尿外科重庆 400016 
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中文摘要:
      目的:观察多西紫杉醇是否诱导前列腺癌PC-3细胞产生自噬,并初步探讨其可能机制。方法:实验应用MTT法测定细胞的增殖率;透射电镜观察细胞超微结构的变化;免疫荧光和Western blot分别定性和定量检测自噬相关蛋白Ⅱ型微管相关蛋白轻链3蛋白(microtubule-associated protein 1 light chain 3 typeⅡ,LC3-Ⅱ)、底物蛋白(sequestosome 1/SQSTM1,P62)表达水平,RT-PCR检测自噬相关基因Beclin-1 mRNA。结果:MTT结果显示多西紫杉醇能有效抑制PC-3细胞增殖(F=58.684,P=0.000);10-6 mol/L多西紫杉醇处理PC-3细胞24 h,即可发生明显的自噬,透射电镜观察到自噬体双层膜结构的形成。免疫荧光和Western blot结果显示多西紫杉醇处理PC-3细胞后P62表达降低(P=0.003、P=0.000、P=0.000),LC3-Ⅱ表达增强(P=0.002、P=0.000、P=0.000),RT-PCR结果显示多西紫杉醇处理PC-3细胞后可引起细胞Beclin-1 mRNA水平的上调(P=0.000、P=0.000、P=0.000)。结论:多西紫杉醇可以有效抑制前列腺癌PC-3细胞的增殖并诱导自噬,其发生可能与通过上调Beclin-1的mRNA水平有关。
英文摘要:
      Objective:To investigate the autophagy induced by docetaxel in prostate cancer cell line PC-3 and to explore its possible mechanism. Methods:PC-3 cells cultured in vitro were treated with docetaxel. Proliferation inhibition rate of PC-3 cells was assayed by MTT;ultra-structural cellular changes were observed by transmission electron microscope(TEM);levels of sequestosome 1/SQSTM1 (P62) and microtubule-associated protein 1 light chain 3 typeⅡ(LC-3Ⅱ) protein were detected by Immunofluorescence and Western blot,qualitatively and quantitatively;level of mRNA of autophagy gene Beclin-1 was detected by RT-PCR. Results:After docetaxel treatment,proliferation of PC-3 cells was inhibited effectively(F=58.684,P=0.000). Autophagy could be observed in prostate cancer PC-3 cells treated with 10-6 mol/L docetaxel after 24 h. TEM examination revealed the appearance of autophagosome with double-membrane structure. Immunofluorescence and Western blot showed that expression of LC-3Ⅱ was increased(P=0.002,P=0.000,P=0.000) in PC-3 cells after being treated with docetaxel and expression of P62 was decreased(P=0.003,P=0.000,P=0.000). RT-PCR showed that levels of mRNA of Beclin-1 were increased by docetaxel(P=0.000,P=0.000,P=0.000). Conclusion:Docetaxel could inhibit the growth of PC-3 cells and induce the appearance of autophagy in PC-3 cells,and the possible mechanism of autophagy induced by docetaxel is related to the increase of Beclin-1 mRNA.
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