茶多酚联合顺铂对人卵巢癌细胞SKOV3生长的影响及机制  

Effect of tea ployphenols combined with cisplatin on SKOV3 ovarian cancer cell line and the mechanism

作　　者：王倩[1] 张蓓[2] 陆晓媛[3] 

机构地区：[1]徐州医学院研究生院 [2]东南大学医学院附属徐州市中心医院妇产科 [3]徐州医学院附属医院妇产科

出　　处：《中华临床医师杂志（电子版）》2012年第14期3937-3941,共5页Chinese Journal of Clinicians(Electronic Edition)

摘　　要：目的观察茶多酚(TP)、顺铂(DDP)及二者联合对人卵巢癌细胞SKOV3增殖及凋亡的影响,初步探讨二者联合对SKOV3细胞生长影响的机制。方法人卵巢癌细胞SKOV3经低毒剂量TP、DDP单独或联合作用后,MTT法检测细胞增殖情况,DAPI核染色法荧光显微镜观察细胞凋亡形态变化,AnnexinV-FITC双染流式细胞术分析细胞凋亡,Westernblot方法检测细胞中Akt及p-Akt的表达。结果低毒剂量TP单药组、DDP单药组与联合用药组对细胞的增殖抑制率分别为(11.47±2.07)%、(32.26±4.85)%、(52.62±3.23)%,联合用药组的细胞增殖抑制率明显高于DDP单药组,差异有统计学意义(P=0.000);荧光显微镜下可见低毒剂量TP单药组的细胞核形态及染色与阴性对照组无明显差异,联合用药组细胞核比DDP单药组着色更重,核浓缩、核碎裂现象更加明显,呈现典型的细胞凋亡征象;低毒剂量的TP对细胞的凋亡无明显影响,联合用药组的凋亡率明显高于DDP单药组,差异有统计学意义(P=0.000);各用药组细胞中总的Akt蛋白表达无明显变化,p-Akt蛋白表达降低,其中联合用药组细胞中p-Akt蛋白表达降低最明显,与对照组及单药组比较,差异均有统计学意义(P<0.001)。结论 TP与DDP联合后可显著增强DDP的抑制细胞增殖、促细胞凋亡效应,可能是通过抑制Akt蛋白的磷酸化来实现的。Objective To study the effect and mechanism of tea ployphenols(TP),cisplatin(DDP)alone and in combination upon the proliferation and cell apoptosis of human ovarian cancer cells SKOV3 in vitro.Methods SKOV3 cells were treated with 6.25 μg/ml TP,2.5 μg/ml DDP,and 6.25 μg/ml TP plus 2.5 μg/ml DDP.Cell proliferation was determined by MTT assay,the morphology of apoptotic cells was observed by DAPI staining,and Annexin V-FITC/PI double staining flow cytometry was used to analyze apoptosis.The expression of Akt and Akt phosphorylation were detected by Western Blot.Results The inhibiting rates of cells treated with the single TP,DDP,and TP plus DDP for 48 h were(11.47±2.07)%,(32.26±4.85)% and(52.62±3.23)%,respectively;the antiproliferation effect of TP combined with DDP group was superior to single DDP group(P=0.000).The results of DAPI staining showed that apoptotic changes including karyorrhexis and karyopyknosis appeared after treatment of single DDP and in combination with TP,and the apoptotic changes in TP combined with DDP group was more obvious than those in single DDP group.Low doses of TP had no significant apoptosis-promoting effect,but the combined treatment group apoptosis rate was significantly higher than the DDP single group.Western blot analysis showed that single groups and TP combined with DDP both unchanged the expression of total Akt protein in human ovarian cancer SKOV3 cells,reduced the expression of Akt phosphorylation,and the effect was more obvious in TP combined with DDP group.Conclusions TP combined with DDP can synergistically inhibit the proliferation of SKOV3 ovarian cancer cell line,induce cell apoptosis,which may be associated with the reduction of AKT phosphorylation.

关 键 词：顺铂 卵巢肿瘤 茶多酚 AKT 

分 类 号：R737.31[医药卫生—肿瘤]