Evn-50增强人卵巢癌顺铂耐药株COC1/DDP对顺铂敏感性的研究  

Evn-50 increase the sensitivity of human ovarian cancer cell line COC1/DDP to cisplatin in vitro

作　　者：蒋静[1] 谢宛玉[1] 曹建国[2] 白军[3] 刘登辉[4] 

机构地区：[1]南华大学第一附属医院妇产科,湖南衡阳421001 [2]湖南师范大学药物工程实验室 [3]杭州市红十字会医院妇产科 [4]南华大学第一附属医院肿瘤外科,湖南衡阳421001

出　　处：《中华临床医师杂志（电子版）》2013年第2期126-129,共4页Chinese Journal of Clinicians(Electronic Edition)

摘　　要：目的观察黄荆子乙酸乙酯提取物(Evn-50)和顺铂(DDP)在体外对人卵巢癌耐顺铂细胞株COC1/DDP凋亡的影响,探讨Evn-50提高人卵巢癌耐顺铂细胞株COC1/DDP对DDP敏感性的机制。方法运用MTT、FCM、Hoechst33258染色法、Westernblot等方法,观察Evn-50增强DDP抑制COC1/DDP细胞增殖作用和此过程中COC1/DDP细胞凋亡,caspase-3蛋白、增殖细胞核抗原(PCNA)及其泛素化修饰蛋白(Ubi-PCNA)表达的改变。结果 Evn-50不仅增强DDP对COC1/DDP细胞的抑制增殖作用,也增强DDP对COC1/DDP细胞的凋亡作用,同时caspase-3蛋白表达增加(P<0.05);而Evn-50(20μg/ml)组与空白对照组相比PCNA蛋白表达下调(P<0.05),Evn-50与DDP合用组与DDP单药组均出现了Ubi-PCNA蛋白表达,Evn-50与DDP合用组与DDP组比较,Ubi-PCNA蛋白表达下调(P<0.05)。结论 Evn-50可能通过减少PC-NA表达,使DDP诱导的PCNA泛素化水平下降,从而逆转DDP耐药,最终通过激活caspase-3蛋白高表达,启动caspase级联反应,增强DDP对人卵巢癌DDP耐药细胞株COC1/DDP的凋亡作用。Objective To investigate the effects of the acetoacetate extract of Vitex negundo seed（Evn-50） and cisplatin（DDP） on the inducing apoptosis of ovarian carcinoma cell line COC1/DDP in vitro and the potential mechanism of increased response rate of DDP induced by Evn-50. Methods To evaluate Evn-50 increased the inhibitory effect of DDP on the proliferation of COC1/DDP by the MTr assay and the apoptotic effect by Hoechst33258 staining and flow cytometric analysis. The expressions of caspase-3, proliferating cell nuclear antigen （PCNA） ,Ubi-PCNA were analyzed by Western blot. Results Evn-50 increase the inhibitory effect of DDP on the proliferation of COC1/DDP and the effect of DDP on inducing apoptosis and the expression of caspase-3 protein（P 〈 0. 05）. Yet,the expression of PCNA protein decreased with Evn-50 used only and increased obviously with control （P 〈 0. 05）. Ubi-PCNA expressed when DDP used only and Evn-50 and DDP, the expression of Ubi-PCNA protein decreased with Evn-50 and DDP and increased obviously with DDP used only （ P 〈 0. 05 ）. Conclusions Evn-50 reduce Ubi-PCNA expression guided by DDP through downregulation of PCNA, thus reverse resistance of DDP. Finally,the same of apoptosis of COC1/DDP is induced by low concentration DDP and high con-centration DDP, through active caspase cascade.

关 键 词：卵巢肿瘤 顺铂 细胞凋亡 Evn-50 逆转耐药 

分 类 号：R737.31[医药卫生—肿瘤]