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  

Effect of compound Chinese traditional medicine PC-SPES Ⅱ in inhibiting proliferation of human prostate cancer cell LNCaP and on expressions of AR and PSA

作　　者：张碧严 李玉凤[1] 赖芸[1] 李云森[2] 陈子珺[1] 

机构地区：[1]上海中医药大学中药学院,上海201203 [2]苏州大学生物医学研究院,江苏苏州215006

出　　处：《中国中药杂志》2015年第5期950-956,共7页China Journal of Chinese Materia Medica

基　　金：上海市博士学位点建设科研项目(K110404);国家"重大新药创制"科技重大专项(2012ZX09401-014)

摘　　要：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LNCa P增殖的作用。采用MTT法检测PC-SPESⅡ对LNCa P细胞增殖的影响,显示PC-SPESⅡ质量浓度为180-1 440 mg·L-1时能显著抑制LNCa P细胞增殖,PC-SPESⅡ作用24,48 h的IC50分别为311.48,199.01 mg·L-1;流式细胞仪检测细胞周期分布的变化发现240 mg·L-1PCSPESⅡ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Hoechst 33258染色观察凋亡细胞形态和Annexin V-FITC/PI双染流式细胞仪检测凋亡细胞比例,PC-SPESⅡ质量浓度为480 mg·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LNCa P细胞分泌PSA的水平,以25 mg·L-1Bic作为阳性对照药,发现480 mg·L-1PC-SPESⅡ能显著降低细胞分泌PSA;采用qRT-PCR和Western blot方法检测AR,PSA mRNA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LNCa P细胞后,240-480 mg·L-1PC-SPESⅡ能显著下调AR,PSA mRNA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LNCa P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。To investigate the effect of compound Chinese traditional medicine PC-SPES lI I in inhibiting proliferation of human prostate cancer cell LNCaP based on the androgen receptor （AR） signaling pathway. The effect of PC-SPES Ⅱ on LNCaP cell prolifer- ation was detected by MTT assay. According to the findings, at the mass concentration of 180-1 440 mg·L-1 , PC-SPES Ⅱ signifi- cantly inhibited the proliferation of LNCaP cells ; the ICs0 of PC-SPES Ⅱ at 24 h and 48 h were 311.48, 199.01 mg·L^-1, respective- ly. The flow Cytometry detection showed 240 mg·L^-1 PC-SPES Ⅱ arrested cells in GJM phase, and an obvious apoptotic peak ap- peared before G0/G1 peak and rose over time. Meanwhile, Hoechst 33258 staining revealed apoptotic cellular morphology. Annexin V-FITC/PI staining manifested an increase in apoptotic cell ratio at the PC-SPES Ⅱ concentration of 480 mg·L^-1 in a dose dependent manner. The prostate specific antigen （PSA） secretion of LNCaP cells was tested by PSA ELISA kit. Besides, compared with 25 mg ·L^-1 Bic, 480 mg·L^-1 PC-SPES Ⅱ significantly reduced the cell secretion of PSA. The AR and PSA mRNA and protein expressionswere detected by qRT-PCR and Western blot. According to the results, after the induction of LNCaP cells with synthetic androgen 25 μg·L^-1 R1881, 240-480 mg·L^-1 PC-SPES Ⅱ notably down-regulated the AR and PSA mRNA and protein expressions and inhibited the translocation of AR from cytoplasm to nucleus. In summary, PC-SPES Ⅱ significantly can inhibit the in vitro proliferation of LNCaP cells and arrest cell cycle arrest in G2/M phase. Its mechanism may be associated with the down-regulation of the AR and PSA expres- sions and the inhibition of AR nuclear translocation.

关 键 词：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 

分 类 号：R285[医药卫生—中药学]