蛋白激酶B抑制剂哌立福辛对卵巢透明细胞癌ES2细胞系的杀伤作用  

Effects of protein kinase B inhibitor Perifosine on ovarian clear cell carcinoma ES2 cell line in vitro

作　　者：吕昌帅[1] 张英兰[1] 周星楠 郎景和[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院妇产科,北京100730

出　　处：《基础医学与临床》2016年第3期375-379,共5页Basic and Clinical Medicine

摘　　要：目的探讨蛋白激酶B抑制剂哌立福辛对卵巢透明细胞癌ES2细胞系的杀伤作用。方法体外培养人卵巢透明细胞癌ES2细胞,以不同药物浓度(0、7、9、11、13和15μmol/L)哌立福辛处理细胞24 h,用细胞计数试剂盒CCK-8检测细胞的存活率;DAPI染色观察细胞核的形态;用annexin V-FITC/PI凋亡检测试剂盒及流式细胞术检测细胞的凋亡;实时荧光定量PCR法检测Ki-67和MCM2 mRNA表达。结果哌立福辛能够降低细胞存活率(P<0.05),在一定程度上呈浓度依懒性;哌立福辛作用于细胞后,荧光显微镜下可见大量的核碎裂及凋亡小体;随着哌立福辛浓度的增加,细胞的凋亡率增加(P<0.01);药物作用48 h后,Ki-67和MCM2 mRNA表达水平明显降低(P<0.01)。结论哌立福辛对人卵巢透明细胞癌ES2细胞有明显的杀伤作用,并且可以负性调节Ki-67和MCM2mRNA的表达。Objective To investigate the effects of protein kinase B inhibitor Perifosine on ovarian clear cell car- cinoma ES2 cell line in vitro. Methods The human ovarian clear cell carcinoma ES2 cells were cultured and in- cubated with different doses of Perifosine（0,7,9,11,13,15 μmol/L） for 24 h. Survival rate of ES2 cells was evaluated with cell counting kit-8 （ CCK-8 ）. Morphology of nuclei were observed with DAPI staining. Cell apop- tosis was analyzed with annexin V-FITC/PI apoptosis kit for flow cytometry. The expression levels of Ki-67 and MCM2 mRNA were detected by RT-PCR. Results The survival rate decreased in ES2 cells treated by Perifos- ine after 24 h（P 〈 O. 05 ） and the rate was related with concentration to a certain extent. A large number of nuclear fragmentation and apoptotic bodies were observed under fluorescence microscope after treatment by Per- ifosine. Cell apoptosis rate increased with the higher concentration of Perifosine（ P 〈 0.01 ）. The expression of Ki-67 and MCM2 mRNA was significantly decreased after 48 h （ P 〈 0.01 ）. Conclusions Perifosine has dis- tinctive inhibitory effects on human ovarian clear cell carcinoma ES2 cells, and is able to negatively regulatethe expression of Ki-67 and MCM2 mRNA.

关 键 词：卵巢透明细胞癌ES2细胞系 哌立福辛 KI-67 MCM2 细胞凋亡 

分 类 号：R737.31[医药卫生—肿瘤]