CAJ | 학술논문

目的探讨姜黄素(Cur)对具有多药耐药特性的人结直肠癌耐奥沙利铂(Oxa)细胞株耐药的逆转作用及其是否与下调耐药蛋白P-糖蛋白(P-gp)有关.方法采用WST-1试剂检测Cur对两种人结直肠癌耐Oxa细胞株(SW480/OxR及SW620/OxR)的半数抑制浓度(IC50),选择比IC50低一浓度梯度的Cur+2μmol/L Oxa作用于两种耐药细胞48h(称为实验组),与仅加入2μmol/L的Oxa对照组比较,采用流式细胞术检测细胞凋亡;采用实时定量聚合酶链反应(Real-time PCR)检测编码P-gp的多药耐药基因-1(MDR-1) mRNA表达;采用流式间接法检测P-gp阳性细胞百分率.结果实验组与对照组比较:(1) SW480/OxR细胞凋亡率由(4.23±1.65)％上升到(32.15±3.98)％,SW620/OxR细胞凋亡率由(5.08±1.82)％上升到(30.69±2.94)％,两种耐药细胞实验组细胞凋亡率均明显高于其对照组(P＜0.05);(2)两种耐药细胞MDR-1 mRNA表达抑制率均超过50％(P＜0.05);(3)两种耐药细胞P-gp蛋白阳性细胞百分率明显下降(P＜0.05).结论 Cur具有逆转结直肠癌耐药细胞株耐药的作用,可能与下调耐药蛋白P-gp表达有关.
목적 탐토강황소(Cur)대구유다약내약특성적인결직장암내오사리박(Oxa)세포주내약적역전작용급기시부여하조내약단백P-당단백(P-gp)유관.방법 채용WST-1시제검측Cur대량충인결직장암내Oxa세포주(SW480/OxR급SW620/OxR)적반수억제농도(IC50),선택비IC50저일농도제도적Cur+2μmol/L Oxa작용우량충내약세포48h(칭위실험조),여부가입2μmol/L적Oxa대조조비교,채용류식세포술검측세포조망;채용실시정량취합매련반응(Real-time PCR)검측편마P-gp적다약내약기인-1(MDR-1) mRNA표체;채용류식간접법검측P-gp양성세포백분솔.결과 실험조여대조조비교:(1) SW480/OxR세포조망솔유(4.23±1.65)％상승도(32.15±3.98)％,SW620/OxR세포조망솔유(5.08±1.82)％상승도(30.69±2.94)％,량충내약세포실험조세포조망솔균명현고우기대조조(P＜0.05);(2)량충내약세포MDR-1 mRNA표체억제솔균초과50％(P＜0.05);(3)량충내약세포P-gp단백양성세포백분솔명현하강(P＜0.05).결론 Cur구유역전결직장암내약세포주내약적작용,가능여하조내약단백P-gp표체유관.
Objective To investigate the reversal multidrug resistance effects of curcumin on human colorectal cancer cell lines resistant to oxaliplatin and whether its mechanism is involved in downregulation of the expression of P-glycoprotein (P-gp).Methods The 50％ inhibitory concentration (IC50) values of curcumin to the two colorectal cancer cell lines resistant to oxaliplatin (SW480/OxR and SW620/ OxR) were obtained by WST-1 reagent,which is one kind of cell proliferation assay.Later experiments included in the experimental group and the control group.In the experimental group,the two kinds of drug-resistant cells were exposed to the previous experiment IC50 concentrations of curcumin plus 2 μmol/L oxaliplatin for 48 h,while in the control group,the two kinds of drug-resistant cells were cultured in medium containing in 2 μmol/L oxaliplatin.In the two groups of the two kinds of drug-resistant cells:apoptosis was detected by flow cytometrythe mRNA expression levels of multidrug resistance (MDR)-1 were detected by real time-polymerase chain reaction (Real-time PCR) and the percentage of P-gp positive cells was disclosed by indirect method of flow cytometry.Results In the control group,the apoptosis rate of SW480/OxR and SW620/OxR cells were respectively (4.23 ± 1.65) ％ and (5.08 ± 1.82) ％,while the apoptosis rate of the two drug-resistant cells in their experimental group respectively rose to (32.15 ± 3.98) ％ and (30.69 ± 2.94) ％.The apoptosis rate of two drug-resistant cells in the experimental group were significantly higher than that of their control group (P ＜ 0.05).In the experimental groups,the mRNA expression of MDR-1 of the two drug-resistant cells decreased more than 50％ (P ＜ 0.05),and the percentage of positive cells of P-gp protein were significantly decreased (P ＜ 0.05),when compared with that in their control group.Conclusion Curcumin can reverse drug resistance in colorectal cancer cell lines resistant to oxaliplatin,its effect may be associated with lowered P-gp expression.