【摘要】 目的:探讨针对肺耐药基因(LRPgene)设计的反义寡核苷酸(AsODN)对人卵巢癌多药耐药细胞系OVCAR-3顺铂(DDP)耐药性的逆转作用。方法:采用罗丹明B(SRB)显色法检测LRPAsODN和DDP细胞毒性作用;采用脂质体(lipofectamine2000)介导LRPAsODN转染人卵巢癌多药耐药细胞系OVCAR-3;采用逆转录酶链式反应(RT-PCR)和Westernblot法检测LRPAsODN转染前后OVCAR-3细胞中LRP的表达;采用流式细胞仪测定OVCAR-3细胞在顺铂作用后的凋亡率。结果:LRPAsODN浓度低于800nmol/L时,对OVCAR-3细胞无明显细胞毒作用;LRPAsODN能明显减少OVCAR-3细胞中LRPmRNA和蛋白的表达,增加其顺铂敏感性,且与剂量呈正相关;转染LRPAsODN前后的OVCAR-3细胞在DDP作用后,凋亡率分别为(15.43±1.14)%和(27.43±2.05)%,两者间差异有显著性意义(P=0.001)。结论:LRPAsODN能有效阻断OVCAR-3细胞内LRP的表达,恢复细胞对DDP的敏感性。更多还原

【Abstract】 Objective: To evaluate the effect of antisense oligonucleotide(AsODN), which was targeted at LRP gene, on reverting cisplatin-resistance of multidrug-resistant human ovarian cancer cell line OVCAR-3. Methods: The cytotoxicity of LRP AsODN and cisplatin, or comined both of them were assayed by sulforhodanmine B(SRB) method. LRP AsODN was transfected into OVCAR-3 cell by lipofectamine 2000. The expression of LRP was monitored by RT-PCR and Western blot in LRP AsODN transfected and non-transfected OVCAR-3 cells and the apopototic rates of both cells were detected by flowcytometry after having been treaded with cisplatin. Results: The cytotoxity of LRP AsODN was very low when its concentration was under 800nmol/L. The expression of LRP mRNA and protein were inhibited and a positive drug resistance to cisplatin was observed after LRP AsODN was transfected into OVCAR-3 cells. The apopototic rates of LRP AsODN transfected and non-transfected cells were (27.43±2.05)% and (15.43±1.14)% respectively, the difference was statistically significant(P=0.001). Conclusion: Transfection of LRP AsODN could interrupt the expression of LRP and restore drug sensitivity to cisplatin in multidrug-resistant human ovarian cancer cell line OVCAR-3.更多还原