SiRNA干扰结肠癌转移相关基因1表达对卵巢癌顺铂化疗耐药性的影响  

Effect of siRNA targeting MACC1 gene on chemoresistance to cisplatin in ovarian cancer

作　　者：李霞[1] 史惠蓉[1] 邓佑兴 张瑞涛[1] 

机构地区：[1]郑州大学第一附属医院妇产科,河南郑州450052

出　　处：《肿瘤》2014年第1期7-12,共6页Tumor

基　　金：河南省重点科技攻关计划项目(编号:122102310545)

摘　　要：目的:利用RNA干扰技术抑制结肠癌转移相关基因1(metastasis-associated in colon cancer 1,MACC1)在顺铂(cisplatin,DDP)耐药的卵巢癌细胞株SKOV-3/DDP中的表达,并探讨其对DDP化疗耐药性的影响。方法:RNA干扰分为3组,空白对照组为未经处理的SKOV-3/DDP细胞,阴性对照组为空质粒转染的SKOV-3/DDP-shVector细胞,实验组为MACC1-shRNA重组质粒转染的SKOV-3/DDP-shMACC1细胞。反转录PCR法和蛋白质印迹法检测稳定转染后SKOV-3/DDP细胞中MACC1 mRNA和蛋白的表达情况,MTT法检测细胞对DDP的耐药性变化,FCM法检测细胞凋亡变化。结果:相比于未转染组SKOV-3/DDP细胞,MACC1-shRNA转染组SKOV-3/DDP-shMACC1细胞中MACC1的mRNA及蛋白水平均明显下调[下调百分率分别为(83.39±1.26)%和(82.25±2.94)%,P<0.05]。DDP对SKOV-3/DDP-shMACC1细胞的半数抑制浓度(half inhibitory concentration,IC_(50))值与未转染组和空质粒转染组细胞的IC_(50)值比较,均明显降低[(26.09±0.91)μmol/L vs(47.50±0.40)μmol/L和(47.08±0.45)μmol/L,P<0.05]。SKOV-3/DDP-shMACC1细胞凋亡率明显提高[(13.77±0.60)%vs(3.63±0.30)%和(4.23±0.30)%,P<0.05]。结论:RNA干扰抑制MACC1基因表达能增强卵巢癌耐药细胞SKOV-3/DDP对DDP的敏感性。Objective： To knock down the expression of metastasis-associated in colon cancer-1 （MACC1） gene in drug-resistant ovarian cancer cell line SKOV3/cisplatin （DDP） by RNA interference technology, and to investigate its effect on chemoresistance to DDP. Methods： Ovarian cancer SKOV3/DDP cells were not transfected （blank group, named as SKOV3/DDP cells）, or transfected with empty plasmid （negative control group, named as SKOV-3/DDP-shVector cells） or combinant plasmid containing MACC1 specific small hairpin RNA （shRNA） （experimental group, named as SKOV-3/DDP-shMACC1 cells）. The mRNA and protein expressions of MACC1 were detected by reverse transcription-PCR （RT-PCR） and Western blotting, respectively. The half inhibition concentration （ICs0） of DDP was determined by MTT assay. The apoptosis was examined by flow cyometry. Results： After transfection of MACC1 shRNA into SKOV-3/DDP cells, the expressions of MACC1 mRNA and protein obviously decreased by （83.39±1.26）% and （82.25±2.94）%, respectively, as compared with those of the blank group （P 〈 0.05）. Compared with the blank group and the negative control group, the value of ICs0 of DDP in SKOV-3/DDP-shMACC1 cells significantly decreased [（26.09±0.91） iJmol/L vs （47.50±0.40） μmol/L and （47.08±0.45） μmol/L, P 〈 0.05]. The apoptotic rate of SKOV-3/DDP-shMACC1 cells was significantly higher than those in the SKOV-3/DDP-shVector cells and the SKOV-3/DDP cells [（13.77±0.60）% vs （4.23±0.30）% and （3.63±0.30）%, P 〈 0.05]. Conclusion： The sensitivity to DDP in ovarian cancer SKOV-3/DDP cells can be enhanced by inhibition of MACC1 gene expression induced by RNA interference.

关 键 词：卵巢肿瘤 RNA干扰 抗药性 肿瘤 顺铂 结肠癌转移相关基因1 

分 类 号：R737.31[医药卫生—肿瘤]