miR-199a通过MDR1/P-gp调节肝癌细胞的耐药  

The effect of miR-199a on drug resistance of hepatocellular carcinoma cells by regulating MDR1/P-gp expression

作　　者：戴海峰[1] 梁军[2] 刘红霞[3] 廖国清[4] 

机构地区：[1]江苏省肿瘤医院镇痛科,南京210009 [2]北京军区联勤部药品仪器检验所化学室,北京100071 [3]北京中医药大学护理学院,北京100102 [4]中国人民解放军第309医院肿瘤内科,北京100091

出　　处：《中国医学前沿杂志（电子版）》2013年第11期30-34,共5页Chinese Journal of the Frontiers of Medical Science(Electronic Version)

摘　　要：目的通过检测Hep G2及Hep G2/ADM细胞中miR-199a和MDR1/P-gp表达的差异,探讨miR-199a与肝癌化疗耐药的关系。方法利用Lipofectamine 2000转染寡核苷酸探针,荧光定量PCR方法检测miR-199a和MDR1的表达水平,Western印迹检测P-gp蛋白表达水平,MTT检测细胞存活率。结果与Hep G2细胞相比,Hep G2/ADM细胞中miR-199a表达显著下降MDR1/P-gp表达显著升高。过表达miR-199a能显著增加阿霉素对Hep G2/ADM细胞的抑制率,并抑制MDR1/P-gp的表达。干扰miR-199a能够显著地降低阿霉素对Hep G2细胞的抑制率,并增加MDR1/P-gp的表达。结论 miR-199a通过调节MDR1/P-gp的表达参与肝癌耐药形成,miR-199a可能成为逆转肝癌耐药的作用靶点。Objective To explore the relationship between miR-199a and drug resistance in hepatocellular carcinoma treatment by detecting the expression of miR-199a and MDR1/P-gp in Hep G2 cell line and Hep G2/ADM cell line. Methods Oligonucleotide probes were transfected in Hep G2 and Hep G2/ADM cells using Lipofectamine 2000. Realtime-PCR was used to determine the expression of miR-199a and MDR1. The protein level of P-gp was analyzed by western blot. The viability of Hep G2 and Hep G2/ADM cells was evaluated by MTY assay. Results MiR-199a expression was signiifcantly decreased but MDR1/P-gp expression was significantly increased in Hep G2/ADM cells than that in Hep G2 cells. Overexpression of miR-199a can significantly increase the inhibition rate of adriamycin on Hep G2/ADM cells and inhibit the expression of MDR1/P-gp. While, knock-down of miR-199a can significantly decrease the inhibition rate of adriamycin on Hep G2 cells and stimulate the expression of MDR1/P-gp. Conclusion miR-199a was involved in the mechanism of drug resistance in leukemia by regulating MDR1/P-gp expression, which may provide a potential novel target for overcoming drug-resistance.

关 键 词：肝癌 抗药性 MiR-199a MDR1/P-GP HEP G2/ADM细胞 

分 类 号：R735.7[医药卫生—肿瘤]