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

发布时间：2018-04-17 18:55

  本文选题：有丝分裂驱动蛋白 + 多西紫杉醇耐药　； 参考：《山东大学学报(医学版)》2017年09期

【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(P0.05)。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(P0.05)。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。
[Abstract]:Objective to investigate the therapeutic effect of methoxy-triphenylmethyl-L-cysteine [S(MeO)TLC] targeting mitogen inhibitor KSPin on docetaxel resistant prostate cancer in vitro.Methods the expression levels of PC3DU145 and PC3R)KSP protein were detected by Western blotting in different cell lines of docetaxel resistant prostate cancer cell line (PC3RX).The effect of S(MeO)TLC on the proliferation of drug-resistant cells was observed by MTT assay and Trypan blue staining assay in the blank control group and PC3 group, and the drug resistant cell line PC3R was selected as the study object, and divided into S(MeO)TLC group and control group by Heochst staining.Flow cytometry and RT-PCR techniques were used to detect the apoptosis of drug resistant cells induced by S(MeO)TLC.Results there was no significant difference in the expression of KSP protein between PC3R cells and PC3 cells (P0.05U .PC3R cells IC50). Compared with PC3 cells (IC50: 106nmol / L), there was no significant difference in the expression of KSP protein in PC3R cells treated with S(MeO)TLC for 24 h. 87.9% of the cells stagnated in mitotic phase after being treated with S(MeO)TLC for 24 hours.72 hours after administration, the mitotic arrest cells were significantly apoptotic.Conclusion docetaxel resistance is not related to the expression of KSP protein. S(MeO)TLC can effectively inhibit the proliferation and induce apoptosis of docetaxel resistant prostate cancer cells.In this process, endogenous and exogenous Caspase dependent apoptotic pathways play an important role, and autophagy may play a synergistic role.
【作者单位】： 山东大学附属省立医院泌尿外科;山东大学齐鲁医院泌尿外科;
【基金】：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)
【分类号】：R737.25
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