Evn-50增强人卵巢癌顺铂耐药株COC1/DDP对顺铂敏感性的研究

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、Western blot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(20 μg/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PCNA表达,使DDP诱导的PCNA泛素化水平下降,从而逆转DDP耐药,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:The aim of our study was to investigate the ef ects of the anti-tumor composition of the acetoacetate extract of Vitex Negundo Seed (EVn-50) on the growth of human cervical carcinoma HeLa cells xenografts in nude mice and its possible molecular mechanism. Methods:Models of human cervical cancer HeLa cells xenografts transplanted subcuta-neously in nude mice were established and randomly divided into 7 groups (each group including 5 nude mice):saline group, Taxol group, EVn-50 group, comp-6 group, comp-7 group, comp-8 group and comp-10 group. The volume and weight of Xe-nograts were observed and compared. The alteration of the weight of nude mice, and the change of serum levels of LDH, ALT, Cr and WBC counts were examined and compared. The apoptotic rate of human cervical carcinoma HeLa cells xenografts was analyzed by FCM. The expressions of P53 and Bcl-2 proteins of HeLa cells xenografts were determined by Western blot-ting. Results:EVn-50 and its fractionated extracts could significantly suppress the increasing volume and weight of human cervical carcinoma HeLa cells xenografts in nude mice models in time-dependent manner, yet had no significant ef ect on the weight of nude mice, the serum levels of LDH, ALT, Cr and WBC were counted. When the xenografts were treated with EVn-50 and its fractionated extracts for 16 days, the apoptotic rate of xenografts cells were significantly increased, and the expression of P53 protein was up-regulated and protein level of Bcl-2 was decreased. Conclusion:EVn-50 and its fractionated extracts could suppress the growth of human cervical carcinoma HeLa cells xenografts in nude mice, which may be related to its pro-motion on xenografts cells apoptosis through down-regulation of Bcl-2 expression and activation of P53 expression.

EVn-50抑制人宫颈癌Hela细胞增殖及诱导凋亡的研究

目的 探讨EVn-50对体外培养的人宫颈癌Hela细胞增殖及凋亡的影响.方法 体外培养人官颈癌Hela细胞,台盼蓝拒染法检测Evn-50对人宫颈癌Hda细胞增殖的影响;平皿克隆形成法、软琼脂克隆形成法测定EV-n50对人官颈癌Hela细胞锚定依赖性生长和锚定非依赖性生长作用的影响;AO/EB染色荧光显微镜观察EVn-50诱导Hela细胞凋亡的形态学改变;DNA凝胶电泳确证Evn-50诱导Hela细胞凋亡作用.结果 EVn-50对体外培养人宫颈癌Hela细胞的增殖具有显著抑制作用,呈浓度依赖性.Evn-50可诱导Hela细胞凋亡,AO/EB染色可见典型凋亡小体;DNA凝胶电泳在Evn-50浓度100μg/nL作用48 h出现典型"梯形"DNA条带.结论 EVn-50具有抑制人宫颈癌Hda细胞增殖并诱导细胞凋亡作用.

EVn-50对人宫颈癌Hela细胞增殖和凋亡的影响

目的:研究EVn-50体外对人宫颈癌Hela细胞增殖及凋亡的影响.方法:体外培养Hela细胞,台盼蓝拒染法检测细胞活力;AO/EB染色荧光显微镜观察Evn-50诱导Hela细胞凋亡形态学改变:DNA凝胶电泳确证EVn-50诱导Hela细胞凋亡作用;PI染色流式细胞仪检测Evn-50诱导Hela细胞凋亡率.结果:Evn-50体外对人宫颈癌Hela细胞的增殖具有显著抑制作用,呈浓度依赖性.Evn-50可诱导Hela细胞凋亡,AO/EB染色可见典型凋亡小体;DNA凝胶电泳在EVn-50浓度100μg/mL作用48 h出现典型"梯形"DNA条带;PI染色流式法显示EVn-50在10、100μg/mL作用Hela细胞48 h后,凋亡率分别为(8.80±0.16)%及(20.93±0.62)%,呈浓度依赖性.结论:EVn-50具有抑制人宫颈癌Hela细胞增殖并诱导细胞凋亡作用.