摘要
目的探讨RNA干扰核糖核苷酸还原酶M2(RRM2)对耐药卵巢癌细胞凋亡及侵袭性的影响。方法将RRM2基因的特异性小干扰(siRNA)转染SKOV3/DDP设为干扰组,SKOV3/DDP细胞、SKOV3/DDP-RRM2非特异性阴性细胞设为空白组、阴性组。检测细胞增殖抑制率,计算RI、耐药转染率,荧光PCR技术检测RRM2基因mRNA表达,并分析siRNA转染对SKOV3/DDP耐药指数、RRM2蛋白的影响,采用Transwell观察SKOV3/DDP细胞侵袭能力。结果空白组、阴性组及干扰组转染率均>90%。DDP对SKOV3/DDP细胞属低度耐药,吉西他滨对SKOV3/DDP细胞仍较为敏感。DDP、吉西他滨对干扰组、阴性组、空白组的DDP对细胞的半数抑制浓度(IC50)值比较,差异有统计学意义(P<0.05),DDP、吉西他滨对干扰组细胞的IC50值显著低于阴性组、空白组(P<0.05)。SKOV3/DDP细胞、SKOV3细胞RRM2蛋白的相对表达量比较,差异有统计学意义(P<0.05),且转染Ⅰ组、Ⅱ组、Ⅲ组相对表达量均显著低于阴性组、空白组,其中以转染Ⅰ组细胞的RRM2蛋白相对表达量下降最显著(P<0.05)。干扰组细胞凋亡率显著高于空白组、阴性组,穿膜细胞数显著低于空白组、阴性组(P<0.05)。结论 siRNA可有效抑制RRM2基因在卵巢癌中的增殖与侵袭性,增加耐药细胞的药物敏感性,尤其是促进DDP诱导的耐药细胞凋亡。
Objective To investigate the influence of RNA-interfered ribonucleotide reductase M2（ RRM2） on apoptosis and invasion of ovarian cancer cells with drug resistant. Methods The specific small interfering（ siRNA） transfected SKOV3/DDP of RRM2 gene were designed as the interference group. SKOV3/DDP cells,SKOV3/DDP-RRM2 non-specific negative cells were designed as the blank group and negative group. The cell proliferation inhibition rate was detected,RI and drug resistant transfection rate were calculated,the expression of RRM2 mRNA was detected by fluorescence PCR. The effect of siRNA transfection on SKOV3/DDP resistance index and RRM2 protein was analyzed,and the invasion ability of SKOV3/DDP cells was observed with Transwell. Results The transfection rate of the blank group,negative group and interference group was 90%. The drug resistance of DDPto SKOV3/DDP cells was low and gemcitabine was sensitive to SKOV3/DDP cells. DDP and gemcitabine of the DDP on cell half inhibitory concentration（ IC50） values showed significant differences among three groups（ P〈0. 05）. IC50 values of DDP and gemcitabine on cells in the interference group were significantly lower than the negative group and blank group（ P〈0. 05）.The relative expression of SKOV 3/DDP cells and SKOV 3 cells RRM 2 protein showed significant differences（ P〈0. 05）,and the relative expression was significantly lower in transfection group I,transfection group II and transfection group III than the negative group and blank group（ P〈0. 05）.The decrease of relative expression of RRM2 protein was the greatest in the transfection group I（ P〈0. 05）. The apoptosis rate was significantly higher in the interference group than the blank group and the negative group,transmembrane cells were fewer than the blank group and the negative group（ P〈0. 05）. Conclusion siRNA can effectively inhibit the proliferation and invasion of RRM2 gene in ovarian cancer. It can increase the drug sensitivity of drug resistant cells,especially cells promoting the apoptosis of drug resistant cells induced by DDP.
出处
《实用临床医药杂志》
CAS
2017年第13期67-71,共5页
Journal of Clinical Medicine in Practice
基金
江苏省徐州中心医院创团队科技项目(XZS201626)
