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中药复方PC-SPESⅡ抑制人前列腺癌细胞LNCaP增殖及其对AR,PSA表达的影响

Keywords: 前列腺癌,PC-SPESⅡ,LNCaP细胞,抑制增殖,雄激素受体,前列腺特异性抗原

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Abstract:

基于雄激素受体(AR)信号通路探讨中药复方PC-SPESⅡ抑制人前列腺癌细胞LNCaP增殖的作用。采用MTT法检测PC-SPESⅡ对LNCaP细胞增殖的影响,显示PC-SPESⅡ质量浓度为180~1440mg·L-1时能显著抑制LNCaP细胞增殖,PC-SPESⅡ作用24,48h的IC50分别为311.48,199.01mg·L-1;流式细胞仪检测细胞周期分布的变化发现240mg·L-1PC-SPESⅡ能阻滞细胞于G2/M期,在G0/G1峰前出现明显的凋亡峰,随作用时间的增加而升高;同时采用Hoechst33258染色观察凋亡细胞形态和AnnexinV-FITC/PI双染流式细胞仪检测凋亡细胞比例,PC-SPESⅡ质量浓度为480mg·L-1时凋亡细胞比率增加,作用效果均呈一定剂量依赖性;通过ELISA法检测LNCaP细胞分泌PSA的水平,以25mg·L-1Bic作为阳性对照药,发现480mg·L-1PC-SPESⅡ能显著降低细胞分泌PSA;采用qRT-PCR和Westernblot方法检测AR,PSAmRNA和蛋白表达的变化,结果显示以人工合成雄激素25μg·L-1R1881诱导LNCaP细胞后,240~480mg·L-1PC-SPESⅡ能显著下调AR,PSAmRNA和蛋白表达,抑制AR由细胞质转移入细胞核。以上结果表明,PC-SPESⅡ对LNCaP细胞体外增殖有抑制作用,阻滞细胞周期于G2/M期并诱导细胞凋亡,其机制可能与下调AR,PSA的表达,抑制AR核转位有关。

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