二甲双胍逆转人卵巢癌细胞株对顺铂耐药的实验研究  

Experimental study of metformin inverses cisplatin-resistance of human ovarian cancer cell line

作　　者：王小霞[1] 尹书琴[1] 康佳丽[1] 袁金[1] WANG Xiaoxia YIN Shuqin KANG Jiali YUAN Jin.(Department of Obstetrics and Gynecology, Guangzhou First Pepole＇s Hospital, Guangzhou 510180, China)

机构地区：[1]广州市第一人民医院妇产科,广州510180

出　　处：《临床肿瘤学杂志》2017年第1期12-16,共5页Chinese Clinical Oncology

摘　　要：目的探讨二甲双胍对人卵巢癌耐顺铂细胞株SKOV3/DDP的耐药逆转作用及机制。方法将人卵巢癌细胞株SKOV3细胞设为亲本组,耐顺铂细胞株SKOV3/DDP细胞分为空白对照组、顺铂组、二甲双胍组和联合用药组(顺铂+二甲双胍组);采用CCK-8法检测不同浓度二甲双胍对SKOV3/DDP细胞的作用,RT-PCR和Western blotting检测各组细胞中内质网应激相关蛋白的表达。结果不同浓度二甲双胍作用24h后,对SKOV3细胞和SKOV3/DDP细胞均有抑制作用,且呈浓度依赖性(P<0.05);当二甲双胍>5 mmol/L时,其对SKOV3/DDP细胞的增殖抑制率高于SKOV3细胞(P<0.05)。顺铂对SKOV3细胞和SKOV3/DDP细胞的半数抑制浓度(IC_(50))分别为15.25μg/ml和118.68μg/ml,SKOV3/DDP的耐药倍数为7.78。联合用药组SKOV3/DDP细胞IC_(50)为73.45μg/ml,耐药倍数为4.81。二甲双胍对SKOV3/DDP细胞对顺铂耐药性的逆转倍数为1.62。亲本组中GRP78 mRNA和蛋白的相对表达水平分别为0.38±0.02和0.24±0.04;顺铂组、二甲双胍组和联合用药组的GRP78 mRNA的相对表达水平分别为0.93±0.02、0.68±0.02、0.49±0.07,与空白对照组的0.83±0.04比较,联合用药组明显降低,其次为二甲双胍组,顺铂组升高,差异均有统计学意义(P<0.05);GRP78蛋白在各组中表达与其mRNA表达趋势一致,差异均有统计学意义(P<0.05)。顺铂组、二甲双胍组、联合用药组CHOP蛋白的表达量分别为0.42±0.03、0.69±0.03、0.84±0.01,均高于空白对照组的0.39±0.05,差异均有统计学意义(P<0.05)。结论二甲双胍对人卵巢癌顺铂耐药细胞的耐药性有调节作用,其可能作用机制为通过降低GRP78表达,提高CHOP蛋白表达的内质网应激途径促进细胞凋亡来降低耐药细胞对顺铂的耐药性。Objective To study the reversal effect and mechanism of metformin on cisplatin resistant human ovarian cancer cell line SKOV3/DDP. Methods The growth inhibitory effects of cisplatin on parental group SKOV3 cell, cisplatin group SKOV3/ DDP cell and combination group SKOV3/DDP cell were detected with CCK-8 assay, and the half maximal inhibitory concentration （IC30）, resistance index and reversal index were calculated. Endoplasmic retieulum stress-related apoptosis related protein expressions were detected by reverse transeription-polymerase chain reaction（RT-PCR） and Western blotting method. Results SKOV3 cells and SKOV3/DDP cells were inhibited by different concentrations of metformin for 24 h, in concentration dependent manner（ P〈0. 05 ）. When mefformin〉5 mmol/L, the proliferation inhibition rate of SKOV3/DDP cells was higher than that of SKOV3 cells （P〈0. 05 ）. The IC50 of cisplatin on parental group SKOV3 cell and cisplatin group SKOV3/DDP cell and combination group SKOV3/DDP cell were 15.25 μg/ml, 118.68 μg/ml and 73.45 μg/ml, respectively.The resistance index of cisplatin group SKOV3/DDP cell and combination group SKOV3/DDP cell were 7.78 and 4. 81 ,respectively.The reversal index of metformin on the resistance of eisplatin was 1.62. The relative expression level of GRP78 mRNA of SKOV3/DDP in cisplatin group, metformin group and combination group were 0. 93 ± 0. 02, 0. 68±0. 02, 0. 49±0. 07. Compared with control group （0. 83±0. 04）, combination group significantly decreased expression, followed by metformin group, cisplatin group was increased （ P〈0.05 ） ; the expression of GRP78 protein level was consistent with mR- NA expression in each group （P〈0. 05）. The expression of CHOP protein in cisplatin group, metformin group and combined medica- tion group were 0. 42±0. 03, 0. 69±0. 03, 0. 84±0. 01, respectively, which were higher than that of the control group （0. 39±0. 05）.The difference was statistically significant （P〈0. 05）. Conclusion Metformin may

关 键 词：卵巢癌 二甲双胍 化疗耐药 逆转 内质网应激 

分 类 号：R737.31[医药卫生—肿瘤]