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目的研究亚致死量陡脉冲对人卵巢癌多药耐药细胞亚株的杀伤及对多药耐药性的调节,并探讨其可能的作用机制。方法以人卵巢癌细胞亲本株(SKOV3)及多药耐药亚株(SKOV3/ADM)为研究对象,采用60Hz陡脉冲电击处理,甲基噻唑基四唑(MTT)法检测两株细胞的电敏感性及化疗敏感性;逆转录多聚酶链反应(RTPCR)法检测耐药亚株中的多药耐药基因(MDR1)mRNA水平,免疫细胞化学法检测其P糖蛋白(Pgp)表达,用透射电镜观察超微结构的改变。结果SKOV3亲本及耐药亚株对陡脉冲电场的电敏感性相近(P=0.642);亚致死量脉冲处理后的耐药亚株出现:化疗敏感性增强,半效抑制浓度(IC50)值及耐药指数(RI)均减低,MDR1mRNA水平无显著改变(P=0.947),Pgp蛋白表达强度显著减弱(P=0.001),细胞出现凋亡改变。结论陡脉冲对SKOV3多药耐药亚株亦可有效杀伤;亚致死量陡脉冲可逆转多药耐药,其可能的机制为增加膜渗透性的同时亦可能诱导并持久改变了耐药相关蛋白的正常构象与功能。
Objective To study the killing effect of sub-lethal steep pulses on multidrug-resistant human ovarian cancer cell subline and the regulation of multidrug resistance, and to explore its possible mechanism. Methods Human ovarian cancer cell line SKOV3 and SKOV3 / ADM were used as the research objects. Electroporation was performed by 60 Hz steep pulse, and the cell viability was measured by methyl thiazolyl tetrazolium (MTT) assay Sensitivity and chemosensitivity. The multidrug resistance (MDR1) mRNA levels in drug-resistant subtypes were detected by reverse transcription polymerase chain reaction (RT-PCR), the expression of Pgp protein was detected by immunocytochemistry, Electron microscopy changes in ultrastructure. Results The susceptibility of SKOV3 parental and drug resistant sub-strains to steep pulsed electric field was similar (P = 0.642). The sub-lethal dose-resolved drug-resistant subtypes showed chemosensitivity, IC50 value and The drug resistance index (RI) decreased, the level of MDR1 mRNA had no significant change (P = 0.947), the expression of Pgp protein decreased significantly (P = 0.001), and the cell apoptosis changed. Conclusions Steep pulses can kill SKOV3 multi-drug resistant sub-strains effectively. Sub-lethal dose of steep pulse can reverse multidrug resistance. Its possible mechanism is to increase membrane permeability and induce and persist the change of resistance-related protein The normal conformation and function.