CAJ | 학술논문

目的探讨去甲斑蝥酸钠(SNCTD)对耐顺铂人肺腺癌细胞系A549/DDP的逆转作用及可能分子机制.方法 (1)采用CCK法筛选出SNCTD对A549/DDP的无毒浓度(即对细胞抑制率＜10%的药物浓度),并检测出顺铂及与无毒浓度SNCTD联合对耐药细胞株的IC50;(2)采用流式细胞仪检测无毒浓度SNCTD对细胞内罗丹明123的蓄积情况;(3)采用反转录-聚合酶链反应(RT-PCR)检测1、2倍无毒浓度SNCTD处理细胞48h后耐药相关基因mdrlmRNA,MRP1 mRNA的表达.结果 (1)SNCTD对A549/DDP的无毒浓度为5μg/ml,与顺铂联合用药降低A549/DDP的耐药性,逆转倍数为1.97;(2)无毒浓度SNCTD处理耐药细胞48h后细胞内罗丹明123荧光强度明显增强(F=36.99,P＜0.05);(3)1、2倍无毒浓度SNCTD处理耐药细胞后mdr1,MRP1mRNA表达明显减低,并具有浓度依赖性.结论 SNCTD对A549/DDP具有耐药逆转作用,其作用机制可能与下调耐药相关基因mdr1,MRP1的表达,影响膜蛋白外排泵功能有关.
목적 탐토거갑반모산납(SNCTD)대내순박인폐선암세포계A549/DDP적역전작용급가능분자궤제.방법 (1)채용CCK법사선출SNCTD대A549/DDP적무독농도(즉대세포억제솔＜10%적약물농도),병검측출순박급여무독농도SNCTD연합대내약세포주적IC50;(2)채용류식세포의검측무독농도SNCTD대세포내라단명123적축적정황;(3)채용반전록-취합매련반응(RT-PCR)검측1、2배무독농도SNCTD처리세포48h후내약상관기인mdrlmRNA,MRP1 mRNA적표체.결과 (1)SNCTD대A549/DDP적무독농도위5μg/ml,여순박연합용약강저A549/DDP적내약성,역전배수위1.97;(2)무독농도SNCTD처리내약세포48h후세포내라단명123형광강도명현증강(F=36.99,P＜0.05);(3)1、2배무독농도SNCTD처리내약세포후mdr1,MRP1mRNA표체명현감저,병구유농도의뢰성.결론 SNCTD대A549/DDP구유내약역전작용,기작용궤제가능여하조내약상관기인mdr1,MRP1적표체,영향막단백외배빙공능유관.
Objective To investigate the reversal effect and the mechanism of sodium norcantharidate(SNCTD)on human lung adenocarcinoma cell line A549/DDP. Methods CKK assay was used to screen out non-toxic concentration (less than 10 percent of cell inhibition ratio) of SNCTD, and to measure the IC50 of cisplatin and IC50 of innoxious concentration SNCTD plus cisplatin in drug-resistant cell line. The accumulation effect of Rh123 was assayed by flow cytometry after treatment with non-toxic concentration of SNCTD. PT-PCR was used to detect the expression of mdr1, MRP1 gene for the drug-resistant cell line treated with non-toxic concentration of SNCTD for 48h. Results (1)The non-toxic concentration of SNCTD was 5μg/ml. SNCTD could decrease drug resistance to cisplatin. The reversal fold was 1.97. (2)The fluorescence intensity of Rh123 in the cells treated with 5μg/ml SNCTD was obviously increased (F=36.99, P＜0.05). (3)The expressions of mdr1, MRP1 gene decreased significantly in a concentration-dependent manner.Conclusions SNCTD could reverse the resistance to cisplatin in A549/DDP cell line. It possibly downregulates the expression of mdr1, MRP1 gene, and inhibits the function of efflux pump of membrance protein.