mdr1反义寡核苷酸增强多药耐药肝癌细胞HepG2/ADM化疗敏感性的实验研究

Experimental research of chemotherapeutic sensitization for multidrug resistance hepatocellular carcinoma cell HepG2/ADM enhanced by mdr1antisense oligonucleotides

目的探讨在体外实验条件下mdr1反义寡核苷酸对多药耐药肝癌细胞株化疗敏感性的影响。方法以肝癌细胞Hep G2/ADM为研究对象,设立mdr1反义寡核苷酸组和空白试剂组作对照,利用脂质体包载肿瘤耐药基因mdr1的反义寡核苷酸进行细胞转染,通过反转录聚合酶链反应(RT-PCR)、免疫印迹实验(Western blotting)分别检测mdr1基因m RNA和P-gp蛋白表达,通过MTT实验检测细胞转染前后对阿霉素(ADM)、顺铂(DDP)和5-氟尿嘧啶(5-FU)的化疗敏感性。结果 Hep G2/AMD肝癌细胞经反义寡核苷酸处理后,mdr1m RNA、P-gp蛋白表达水平均明显降低,对ADM、DDP和5-FU的化疗敏感性明显增强。结论反义寡核苷酸能在体外有效增加肝癌细胞Hep G2/ADM对化疗药物的敏感性。

Objective To study the effect of mdr1 genes antisense oligonucleotides in vitro on chemotherapeutic sensiti-zation for multidrug resistance hepatocellular carcinoma cell lines. Methods The hepatocellular carcinoma cell HepG2/ADM was taken as research object,and mdr1 genes antisense oligonucleotides group and blank control group were established. The cell transfection was conducted by antisense oligonucleotides of liposome entrapment drug resistance gene mdr1. The protein expres-sions of mRNA and P-gp of mdr1 gene were detected by reverse transcription-polymerase chain reaction（RT-PCR） and Western blotting respectively,while the chemotherapeutic sensitization to ADM,DDP and 5-FU before and after cell transfection by MTT. Results After transfecting by antisense oligonucleotides,the protein expression levels of mdr1,mRNA and P-gp in hepatocellu-lar carcinoma cell HepG2/ADM decreased obviously,and the chemotherapeutic sensitization to ADM,DDP and 5-FU increased prominently. Conclusion The antisense oligonucleotides can improve the sensitization of HepG2/ADM to chemotherapy drugs in vitro.