中药复方PC—SPES Ⅱ抑制人前列腺癌细胞LNCaP增殖及其对AR， PSA表达的影响(1)
[摘要] 基于雄激素受体(AR)信号通路探讨中药复方PC-SPES Ⅱ抑制人前列腺癌细胞LNCaP增殖的作用。采用MTT法检测PC-SPES Ⅱ对LNCaP细胞增殖的影响，显示PC-SPES Ⅱ质量浓度为180～1 440 mg·L-1时能显著抑制LNCaP细胞增殖，PC-SPES Ⅱ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-1 PC-SPES Ⅱ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LNCaP细胞分泌PSA的水平，以25 mg·L-1 Bic作为阳性对照药，发现480 mg·L-1PC-SPES Ⅱ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25 μg·L-1R1881诱导LNCaP细胞后，240～480 mg·L-1 PC-SPES Ⅱ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-SPES Ⅱ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。
, http://www.100md.com
[关键词] 前列腺癌;PC-SPES Ⅱ;LNCaP细胞;抑制增殖;雄激素受体;前列腺特异性抗原
[收稿日期] 2014-07-03
[基金项目] 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)
[通信作者] 陈子珺，副教授，硕士生导师，研究方向为中药复方配伍规律和肿瘤免疫药理，Tel：(021)51322187，E-mail：zjchen21@aliyun.com.cn
[作者简介] 张碧严，硕士研究生，研究方向为肿瘤和免疫药理，E-mail：zhang_bi_yan@hotmail.com
Effect of compound Chinese traditional medicine PC-SPES Ⅱ in inhibiting
, 百拇医药
proliferation of human prostate cancer cell LNCaP and
on expressions of AR and PSA
ZHANG Bi-yan， LI Yu-feng， LAI Yun， LI Yun-sen， CHEN Zi-jun
(1. School of Chinese Materia Medica， Shanghai University of Traditional Chinese Medicine， Shanghai 201203， China;
2. Institutes of Biology and Medical Sciences， Suzhou University， Suzhou 215006， China)
, http://www.100md.com
[Abstract] To investigate the effect of compound Chinese traditional medicine PC-SPES Ⅱ 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 proliferation was detected by MTT assay. According to the findings， at the mass concentration of 180-1 440 mg·L-1， PC-SPES Ⅱ significantly inhibited the proliferation of LNCaP cells; the IC50 of PC-SPES Ⅱ at 24 h and 48 h were 311.48， 199.01 mg·L-1， respectively. The flow Cytometry detection showed 240 mg·L-1 PC-SPES Ⅱ arrested cells in G2/M phase， and an obvious apoptotic peak appeared 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 expressions were 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 expressions and the inhibition of AR nuclear translocation., 百拇医药(张碧严 李玉凤 赖芸 李云森 陈子珺)
, http://www.100md.com
[关键词] 前列腺癌;PC-SPES Ⅱ;LNCaP细胞;抑制增殖;雄激素受体;前列腺特异性抗原
[收稿日期] 2014-07-03
[基金项目] 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)
[通信作者] 陈子珺，副教授，硕士生导师，研究方向为中药复方配伍规律和肿瘤免疫药理，Tel：(021)51322187，E-mail：zjchen21@aliyun.com.cn
[作者简介] 张碧严，硕士研究生，研究方向为肿瘤和免疫药理，E-mail：zhang_bi_yan@hotmail.com
Effect of compound Chinese traditional medicine PC-SPES Ⅱ in inhibiting
, 百拇医药
proliferation of human prostate cancer cell LNCaP and
on expressions of AR and PSA
ZHANG Bi-yan， LI Yu-feng， LAI Yun， LI Yun-sen， CHEN Zi-jun
(1. School of Chinese Materia Medica， Shanghai University of Traditional Chinese Medicine， Shanghai 201203， China;
2. Institutes of Biology and Medical Sciences， Suzhou University， Suzhou 215006， China)
, http://www.100md.com
[Abstract] To investigate the effect of compound Chinese traditional medicine PC-SPES Ⅱ 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 proliferation was detected by MTT assay. According to the findings， at the mass concentration of 180-1 440 mg·L-1， PC-SPES Ⅱ significantly inhibited the proliferation of LNCaP cells; the IC50 of PC-SPES Ⅱ at 24 h and 48 h were 311.48， 199.01 mg·L-1， respectively. The flow Cytometry detection showed 240 mg·L-1 PC-SPES Ⅱ arrested cells in G2/M phase， and an obvious apoptotic peak appeared 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 expressions were 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 expressions and the inhibition of AR nuclear translocation., 百拇医药(张碧严 李玉凤 赖芸 李云森 陈子珺)