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山东大学学报（医学版） ›› 2017, Vol. 55 ›› Issue (9): 23-30.doi: 10.6040/j.issn.1671-7554.0.2017.145

• 基础医学 • 上一篇    下一篇

靶向抑制有丝分裂驱动蛋白治疗多西紫杉醇耐药前列腺癌的体外疗效

董伟1,邢乃栋2,吕家驹1,刘帅1,孙亮1,曹庆伟1,董宇昊1,刘钊1,丁森泰1   

  1. 1.山东大学附属省立医院泌尿外科, 山东 济南 250021;2.山东大学齐鲁医院泌尿外科, 山东 济南 250012
  • 收稿日期:2017-02-17 出版日期:2017-09-10 发布日期:2017-09-10
  • 通讯作者: 丁森泰. E-mail:dingsentai@126.com E-mail:dingsentai@126.com
  • 基金资助:
    国家自然科学基金(81202017);山东省重点研发计划(2016GSF201147);济南市科技发展计划(201121060)

The therapeutic efficacy of a potent KSP inhibitor S(MeO)TLC against docetaxel-resistant prostate cancer

DONG Wei1, XING Naidong2, LÜ Jiaju1, LIU Shuai1, SUN Liang1, CAO Qingwei1, DONG Yuhao1, LIU Zhao1, DING Sentai1   

  1. 1. Department of Urology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong, China;
    2. Department of Urology, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China
  • Received:2017-02-17 Online:2017-09-10 Published:2017-09-10

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摘要： 目的 探讨甲氧基-(S)-三苯甲基-L-半胱氨酸［S(MeO)TLC］靶向抑制有丝分裂驱动蛋白(KSP)在体外试验中对多西紫杉醇耐药前列腺癌的治疗效果。 方法 培养建立多西紫杉醇耐药前列腺癌细胞株(PC3R),Western blotting检测不同细胞株(PC3、DU145及PC3R)KSP蛋白表达水平;实验分空白对照组、PC3R组及PC3组,MTT及台盼兰染色细胞活性实验观察S(MeO)TLC抑制耐药细胞增殖效果;以耐药细胞株PC3R为研究对象,实验分为S(MeO)TLC组及对照组,Heochst染色、流式细胞仪及RT-PCR技术检测S(MeO)TLC诱导耐药细胞凋亡的效果。 结果 PC3、DU145、PC3R细胞KSP蛋白表达量差异无统计学意义(P>0.05)。PC3R细胞S(MeO)TLC半数抑制浓度(IC50)为120 nmol/L,与PC3细胞(IC50:106 nmol/L)比较,差异无统计学意义(P>0.05)。PC3R细胞给予S(MeO)TLC作用24 h后,87.9%细胞停滞在有丝分裂期;给药72 h后,有丝分裂期停滞细胞显著凋亡。与对照组相比,S(MeO)TLC组PC3R细胞Caspase-3(t=13.445, P=0.000 2)、Caspase-8(t=9.494, P=0.000 7)、Caspase-9(t=5.198, P=0.007)、PARP(t=19.097, P=0.000 04)及自噬标志指标LC3(t=22.609, P=0.000 02)和Beclin1(t=61.266, P=0.000 000 4)的mRNA量均明显升高。 结论 前列腺癌多西紫杉醇耐药与KSP蛋白表达无关,KSP蛋白靶向抑制剂S(MeO)TLC能够有效抑制多西紫杉醇耐药前列腺癌细胞增殖并诱导其凋亡。在此过程中,内源性和外源性Caspase依赖性的凋亡途径均发挥了重要作用,且自噬可能发挥协同作用。

关键词: 靶向治疗, 有丝分裂驱动蛋白, 多西紫杉醇耐药, 前列腺肿瘤

Abstract: Objective To investigate the therapeutic effect of S(MeO)TLC which targetedly inhibits KSP(Eg5)on docetaxel-resistant prostate cancer in vitro. Methods Docetaxel-resistant PC3 cells(PC3R)were developed by chronic, repeated exposure to docetaxel through gradient culture. IC50 of docetaxel-resistant cells to docetaxel was assessed with MTT assay. KSP expressions of different prostate cancer cell lines(PC3, DU145, PC3R)were examined with Western blotting. The cells were divided into control group, PC3 group and PC3R group, and the anti-proliferative activity of S(MeO)TLC was analyzed with MTT and trypan blue staining. After that, the PC3R cells were divided into 山 东 大 学 学 报 (医 学 版)55卷9期 -董伟,等.靶向抑制有丝分裂驱动蛋白治疗多西紫杉醇耐药前列腺癌的体外疗效 \=-control group and S(MeO)TLC group, and the apoptosis was investigated with Hoechst nuclear staining, flow cytometry and RT-PCR in vitro. Results The expressions of KSP in PC3, DU145, and PC3R were similar, with no statistical difference(P>0.05). There was no significant difference in IC50 between PC3R(120 nmol/L)and PC3(106 nmol/L)(P>0.05). After S(MeO)TLC treatment for 24 hours, 87.9% PC3R cells were in the mitosis phase, most of which went apoptosis after 72 hours of S(MeO)TLC treatment. Compared with the control group, PC3R group had significantly increased expressions of Caspase-3(t=13.445, P=0.000 2), Caspase-8(t=9.494, P=0.000 7), Caspase-9(t=5.198, P=0.007), PARP(t=19.097, P=0.000 04), LC3(t=22.609, P=0.000 02)and Beclin1(t=61.266, P=0.000 000 4). Conclusion The resistance of prostate cancer to docetaxel is not related to KSP expression. S(MeO)TLC, as a new KSP/Eg5 inhibitor, has shown obvious anticancer efficacy in docetaxel-resistant prostate cancer cell lines in vitro by inducing cell apoptosis, in which both endogenous and exogenous caspase-dependent apoptotic pathways play an important role, while autophagy plays a synergistic role.

Key words: Kinesin spindle protein, Docetaxel-resistant, Prostate cancer, Targeted therapy

中图分类号: 

  • R737.25
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