摘要
目的探讨RABEX-5下调表达后,乳腺癌细胞对蒽环类药物和紫杉类药物的敏感性变化。方法构建RABEX-5RNAi慢病毒表达载体,并感染人乳腺癌细胞株MCF-7,使细胞内RABEX-5基因表达沉默,Realtime PCR、Western blot方法检测沉默效应。应用CCK-8方法检测RABEX-5沉默后(MCF-7/RNAi),与其阴性对照人乳腺癌细胞株(MCF-7/vector)相比对化疗药物敏感性的变化情况。结果 RABEX-5RNAi慢病毒表达载体感染MCF-7后,与未干扰组(MCF-7)及阴性对照组(MCF-7/vector)相比,RABEX-5的mRNA和蛋白水平表达下调。与阴性对照组相比,RABEX-5下调表达后,对蒽环类药物表柔比星的化疗敏感性降低,半数致死浓度(IC50)分别为(3.590 0±0.228 69)μg/mL、(1.193 3±0.187 71)μg/mL,差异有统计学意义(P<0.05);而对紫杉类药物多西他赛的敏感性不变,半数致死浓度(IC50)分别为(11.162 7±0.210 26)μg/mL、(10.536 7±0.430 97)μg/mL,差异无统计学意义(P>0.05)。结论 RABEX-5下调表达使人乳腺癌细胞MCF-7对蒽环类药物表柔比星产生耐药,而对紫杉类药物多西他赛敏感,为进一步研究RABEX-5在乳腺癌个体化治疗中的作用提供理论基础。
Objective To investigate the changes in sensitivity to anthracycline and taxanes of human breast cancer ceils after RABEX-5 downregulated. Methods By constructing a lentiviral vector for RNA interference （RNAi） of RABEX-5 gene and transfected into human breast cancer cell line MCF-7 to silence the express of RABEX 5. Real-time PCR and Western blot were used to detect the silencing effect. The changes in sensitivity to chemotherapeutic drugs of MCF-7/RNAi and its negative control cell lines MCF-7/vector were detected by CCK-8 reagent. Results Compared with MCF-7 cell and MCF-7/vector cell, the expression of RABEX-5 mRNA and protein was downregulated in MCF-7/RNAi cells. The sensitivity to epirubicin was reduced after downregulating the expression of RABEX-5, the 50% inhibition concentration （IC^0） of MCF-7/RNAi [（3. 590 04-0. 228 69） μ/mL] was higher than that of MCF-7/Vvector [（1. 193 3±_0. 187 71） μg/mL, P〈0.05] ; while the same effect was not found indocetaxel group, the IC90 were （11. 162 7±0. 210 26） μg/mI, and （10. 536 7±0. 430 97）μg/ml, respectively （P〉 0.05）. Conclusion Downregulation of RABEvX5 can induce chemoresistance to epirubicin in human breast cancer cell MCF-7 ; while its effect on sensitivity to docetaxel is not significant. This study can provide a theoretical basis for future research RABEX-5 in the individualized treatment of breast cancer.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2013年第6期882-885,共4页
Journal of Sichuan University(Medical Sciences)
基金
重庆市教委基金资助项目(No.KJ100317)资助
