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作 者:郑清水[1] 许宁[1] 李晓东[1] 陈少豪 薛学义[1] 魏勇[1] 林云知
机构地区:[1]福建医科大学附属第一医院泌尿外科,福州350005
出 处:《中华实验外科杂志》2015年第6期1294-1296,共3页Chinese Journal of Experimental Surgery
基 金:福建省卫生和计划生育委员会中医药科技资助项目(wzzy201315);福建省教育厅面上资助科技项目(JA12142);福建医科大学第一临床学院学科带头人培养对象及青年骨干教师培养专项基金资助项目(JGG201305)
摘 要:目的 观察吴茱萸碱联合顺铂对前列腺癌PC-3细胞增殖和细胞间缝隙连接通讯(GJIC)的影响.方法 体外培养PC-3细胞,分为顺铂组(5、10、15、20 μmol/L)和顺铂(5、10、15、20 μmol/L)+吴茱萸碱组(0.4μmol/L)作用PC-3细胞.细胞计数试剂盒(CCK-8)法检测细胞增殖率变化;流式细胞术检测细胞凋亡变化;Western blot分别检测间隙连接蛋白43(Cx43)蛋白表达;荧光光漂白恢复技术检测细胞缝隙连接(GJ)功能改变.结果 顺铂组荧光恢复率分别为(41.93±1.30)%、(36.32±2.33)%、(30.81±1.43)%、(22.54±1.70)%,顺铂+吴茱萸碱组分别为(69.86±2.16)%、(50.03±5.00)%、(45.38 ±2.59)%、(34.01±4.01)%.与顺铂组比较,顺铂联合吴茱萸碱组荧光恢复率、细胞凋亡水平、Cx43蛋白表达水平均明显升高(P<0.05).结论 吴茱萸碱可改善前列腺癌PC-3细胞株对顺铂耐药性,增强顺铂对前列腺癌PC-3细胞杀伤作用.Objective To evaluate the effect of the combination of cisplatin and evodiamine on the growth of PC-3 cells and gap junction intercellular communication (GJIC).Methods Human prostate cancer PC-3 cells in logarithmic growth phase were cultured in vitro and then treated with various concentrations of cisplatin (5,10,15,20 μmol/L) or evodiamine (0.4 μmol/L) combined with various concentrations of cisplatin (5,10,15,20 μmol/L).The effects of the proliferation was investigated by cell counting kit-8 (CCK-8) assay.Apoptosis rate,and cell cycle distribution were examined by Annexin V-fluoresceine isothiocyanate (FITC)/propidiun iodide (PI) double staining flow cytometry.The expression of connexin43 (Cx43) protein was detected by Western blotting.The changes in the GJIC function were observed by FRAP.Results Fluorescence recovery rate was (41.93 ± 1.30) %,(36.32 ± 2.33) %,(30.81 ± 1.43)%,and (22.54 ±1.70)% in cisplatin groups,and (69.86 ±2.16)%,(50.03 ± 5.00)%,(45.38 ±2.59)%,and (34.01 ±4.01)% in Csplatin + evodiamine groups with statistically significant differences within groups respectively (P < 0.05 for all).Fluorescence recovery rate,apoptosis rate,and the expression of Cx43 protein in cisplatin + evodiamine groups were significantly increased as compared with cisplatin groups (P < 0.05 for all).Conclusion Evodiamine could recover the gap junction intercellular communication through increasing the expression of Cx43 protein and enhancing the cytotoxicity of cisplatin to prostate cancer PC-3 cells.
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