【摘要】 目的:探讨反义寡脱氧核苷酸(ASODN)抑制多药耐药基因(mdr1)对耐阿霉素(ADM)肝癌细胞HepG2/ADM化疗敏感性的影响。方法:以多药耐药基因mdr1人工合成寡脱氧核苷酸(ODN),其中ODN分为ASODN和正义寡脱氧核苷酸(SODN),并采用脂质体转染技术将ODN与HepG2/ADM共培养,试验分为反义寡脱氧核苷酸(mdr1-ASODN)处理组、正义寡脱氧核苷酸(mdr1-SODN)对照组和空白对照组,通过逆转录聚合酶链式反应和蛋白免疫印迹法观察HepG2/ADM细胞中mdr1mRNA及其蛋白表达情况;MTT法观察HepG2/ADM细胞转染mdr1-ASODN前、后对化疗药物(ADM、顺铂、5-氟尿嘧啶)的敏感性。结果:与mdr1-SODN对照组和空白对照组比较,mdr1-ASODN处理组HepG2/AMD细胞中mdr1mRNA及其蛋白表达均明显降低(P<0.05);与转染前比较,HepG2/ADM细胞的增殖抑制率明显增加(P<0.05)。结论:ASODN能有效抑制mdr1基因表达,并恢复肝癌HepG2/ADM细胞对化疗药物的敏感性。更多还原

【Abstract】 OBJECTIVE:To investigate the effects on chemosensitivity of adriamycin(ADM)-resistant hepatoma carcinoma cells(HepG2/ADM) by inhibition of multidrug resistance 1(mdr1) expression with antisense oligodeoxyribonucleotide(ASODN).METHODS:Oligodeoxyribonucleotide(ODN) of mdr1 gene was synthesized and included ASODN and SODN,then transfected into HepG2/ADM hepatoma carcinoma cells by liposome transfection reagent.HepG2/ADM hepatoma carcinoma cells were divided into mdr1-ASODN treatment group,mdr1-SODN control group and blank control group.RT-PCR and Western blotting were used to detect their mRNA and protein expressions of mdr1.The sensitivity of HepG2/ADM cells before and after transfected by mdr1-ASODN to chemotherapeutic drugs(ADM,cisplatin,5-fluorouracil) was evaluated by MTT assay.RESULTS:Compared with mdr1-SODN control group and blank control group,the mRNA and protein expressions of mdr1 were significantly decreased in mdr1-ASODN treatment group(P<0.05);compared with before transfection,the inhibitory effects of chemotherapeutic agents on the proliferation of HepG2/ADM hepatoma carcinoma cells were increased in vitro(P<0.05).CONCLUSION:ASODN inhibits the expressions of mdr1 and enhances the sensitivity of HepG2/ADM cells to chemotherapeutic agents in vitro.更多还原