摘要
目的:研究凋亡抑制基因Livin为靶向的RNA干扰技术(RNAi)对增加人宫颈癌细胞系SiHa化疗敏感性的作用及价值。方法:用已经构建好的针对Livin基因的短发夹RNA真核表达载体(shLivin),转染宫颈癌SiHa细胞,分为空白对照组(SiHa组)、SiHa-shLivin组和转染空载体的SiHa-shVector组。选用顺铂、紫杉醇、多柔比星作为化疗药物,采用四甲基偶氮唑盐(MTT)法检测加药后3组细胞的增殖情况,得出半数抑制浓度(IC50)值,用流式细胞术检测3组细胞在shLivinIC50的凋亡率进行比较。结果:shLivin重组质粒稳定转染SiHa细胞后,SiHa-shLivin组对顺铂、紫杉醇、多柔比星的IC50值明显低于SiHa组和SiHa-shVector组,差异有高度统计学意义(P<0.001),SiHa组与SiHa-shVector组比较,差异无统计学意义(P>0.05)。用shLivinIC50药物浓度处理细胞,SiHa-shLivin组凋亡率明显高于SiHa-shVector组和SiHa组,差异有高度统计学意义(P<0.001),Si-Ha组与SiHa-shVector组比较,差异无统计学意义(P>0.05)。结论:以Livin基因为靶向的RNA干扰技术可增加化疗药物的敏感性,作为宫颈癌细胞化疗增敏的分子靶点,其基因沉默有望成为宫颈癌基因治疗新手段。
Objective： To explore the effect and value of the short-hairpin RNA （shRNA） targeting Livin （shLivin） in enhancing chemo-sensitivity of cervical cancer cell SiHa. Methods：The divided experimental groups was as follows： control group, SiHa-shLivin group （SiHa cells were transfected with shLivin）and SiHa-shVector group（ SiHa cells were transfected with shVector）. Cisplatin, Taxol and doxorubicin were used as chemotherapy drugs. MTT was performed to assess cell proliferation and half inhibitory concentrations of chemotherapy drugs were calculated. Cell apoptosis was analyzed by flow cytometry. Results：After silencing of Livin gene by shLivin, significant decreases in IC50 of cisplatin, Taxol and doxorubicin were observed in SiHa cells （ P 〈 0. 001 ）. There was no difference in IC50 of all the chemotherapy drugs between control group and SiHa-shVector group（ P 〉 0.05）. Apoptosis rates after treatment with IC50 of drugs were significantly increased （ P〈0. 001 ）, There was no difference in Apoptosis rates between control group and SiHa-shVector group（ P 〉 0.05）. Conclusions： Livin is hopeful to be a molecular target for increasing the chemotherapeutic response of cervical cancer cells. Gene silen- cing of Livin mediated by RNA interference which can increasing the chemotherapeutic response may be a new means of gene therapy for cervical cancer.
出处
《实用妇产科杂志》
CAS
CSCD
北大核心
2012年第8期658-660,共3页
Journal of Practical Obstetrics and Gynecology
基金
湖北省自然科学基金课题(编号:2008CDB137)
关键词
LIVIN
凋亡
基因沉默
宫颈癌
化疗
Livin
Apoptosis
Gene silencing
Cervical cancer
Chemotherapy
