茶多酚诱导卵巢癌SKOV3细胞凋亡及其机制的研究  

Tea polyphenols induce the apoptosis of ovarian cancer SKOV3 cells and its mechanism

作　　者：付春红[1] 张秦[1] 刘锦霞[1] 李云莉[1] 吴元赭[1] 

机构地区：[1]南京大学医学院临床学院(南京军区南京总医院)妇产科

出　　处：《医学研究生学报》2012年第11期1146-1150,共5页Journal of Medical Postgraduates

摘　　要：目的卵巢癌手术、传统化疗和放疗技术虽然有所改进,但患者5年生存率仍不超过20%。已有研究证明茶多酚具有抗肿瘤和抗氧化作用。文中主要探讨茶多酚对卵巢癌SKOV3细胞的增殖抑制作用和诱导SKOV3细胞凋亡的作用及其作用机制。方法采用MTT比色法检测不同浓度的茶多酚对卵巢癌SKOV3细胞增殖抑制作用;应用荧光素标记的膜联蛋白Ⅴ(FITC-AnnexinⅤ)联合流式细胞凋亡定量检测技术(PⅠ双染色)检测茶多酚对SKOV3细胞凋亡发生率的影响,同时应用PI染色技术分析茶多酚对SKOV3细胞分裂周期的影响;利用荧光探针DCFH-DA对细胞内活性氧自由基(reactive oxygenspecies,ROS)水平进行检测。结果茶多酚对SKOV3细胞的生长有明显的抑制作用,不同剂量茶多酚组的抑制率与空白对照组比较有显著差异(P<0.01)。AnnexinⅤ/PⅠ双染色法观察到药物浓度与细胞凋亡发生呈正相关,即随着药物浓度的增加细胞凋亡率增加,在200μmol/L的时候几乎所有的细胞都发生了凋亡。随着药物浓度的增加处于G0/G1期细胞减少,多数细胞阻滞在细胞周期的S期,从而阻断细胞周期向G2/M期进行,当药物浓度达到200μmol/L时,出现明显凋亡峰。利用荧光探针DCFH-DA对细胞内ROS水平进行检测发现随着茶多酚浓度增高细胞内ROS水平显著增高,不同剂量茶多酚组细胞内ROS水平与空白对照组比较有显著差异(P<0.01)。结论茶多酚在体外可剂量依赖性地抑制卵巢癌SKOV3细胞增殖,诱导细胞凋亡,推测ROS水平的增高可能是导致卵巢癌SKOV3细胞凋亡的原因,为筛选治疗卵巢癌的化疗药物提供新的参考依据。Objective Despite advances in surgical,chemotherapeutic and radiotherapeutic techniques for ovarian cancer,its 5-year survival rate is less than 20%.Tea polyphenols have been proved to play an active role in anti-tumor and anti-oxidant processes.This study was to investigate the effects of tea polyphenols in inhibiting the proliferation and inducting the apoptosis of ovarian cancer SKVO3 cells.Methods We determined the effects of tea polyphenols on inhibiting the proliferation and inducing the apoptosis of ovarian cancer SKVO3 cells using MTT and AnnexinⅤ/PI double staining,respectively.We also detected their effect on the cycles of cell division and measured the level of reactive oxygen species（ROS） inside the cells with fluorescent probes.Results Compared with the blank control group,tea polyphenols significantly inhibited the proliferation of the ovarian cancer SKVO3 cells（P0.01）.Annexin/PI double staining revealed a positive correlation between drug concentration and cell apoptosis,which occurred in almost all the ovarian cancer cells at 200 μmol/L.With the increasing of the drug concentration,the cells in the G0/G1 stage were reduced,mostly blocked in the S stage,holding the process toward the G2/M stage,with a distinct apoptosis peak at 200 μmol/L.The level of ROS inside the cells was significantly elevated with the increasing concentration of tea polyphenols as compared with the control group（P0.01）.Conclusion Tea polyphenols in vitro can inhibit the proliferation of ovarian cancer SKVO3 cells and induce their apoptosis in a dose-dependent manner,which suggests that ROS elevation may contribute to the apoptosis of ovarian cancer SKVO3 cells.Our findings have provided some new reference for the screening of chemotherapy drugs for ovarian cancer.

关 键 词：茶多酚 卵巢癌 SKOV3细胞 活性氧自由基 细胞凋亡 

分 类 号：R737.31[医药卫生—肿瘤]