CAJ | 학술논문

目的:研究分泌型卷曲相关蛋白5(secreted frizzled related protein 5,SFRP5)对P-糖蛋白(P-glycoprotein,P-gp)介导的白血病多药耐药性的作用.方法:采用转基因方法构建过表达SFRP5的KG1a/SFRP5细胞,real-time PCR检测MDR1 mRNA表达,Western blot检测细胞P-gp表达.免疫荧光显微镜观察细胞膜表面P-gp表达.流式细胞仪检测细胞内药物浓度.MTT方法检测细胞耐药性.结果:与KG1a细胞及表达绿色荧光蛋白的KG1a/eGFP细胞相比,KG1a/SFRP5细胞中MDR1 mRNA水平显著下降(P<0.01),总P-gp表达水平亦被下调,细胞膜表面P-gp荧光强度减弱,细胞内的罗丹明浓度显著升高(P<0.01),对ADR的IC50显著降低(P<0.01),细胞耐药性下降.结论:SFRP5蛋白表达可以下调MDR1转录及P-gp表达,增加细胞内药物浓度,逆转白血病多药耐药.
목적:연구분비형권곡상관단백5(secreted frizzled related protein 5,SFRP5)대P-당단백(P-glycoprotein,P-gp)개도적백혈병다약내약성적작용.방법:채용전기인방법구건과표체SFRP5적KG1a/SFRP5세포,real-time PCR검측MDR1 mRNA표체,Western blot검측세포P-gp표체.면역형광현미경관찰세포막표면P-gp표체.류식세포의검측세포내약물농도.MTT방법검측세포내약성.결과:여KG1a세포급표체록색형광단백적KG1a/eGFP세포상비,KG1a/SFRP5세포중MDR1 mRNA수평현저하강(P<0.01),총P-gp표체수평역피하조,세포막표면P-gp형광강도감약,세포내적라단명농도현저승고(P<0.01),대ADR적IC50현저강저(P<0.01),세포내약성하강.결론:SFRP5단백표체가이하조MDR1전록급P-gp표체,증가세포내약물농도,역전백혈병다약내약.
Objective:To investigate the effect of secreted frizzled-related protein 5 (SFRP5) on leukemic multidrug resistance. Methods:Transgenic methods were used to culture KG1a cells expressing SFRP5 and enhanced green fluorescent protein (control). mdr1 mRNA expression in KG1a, KG1a/eGFP, and KG1a/SFRP5 groups were detected by real-time polymerase chain reaction. P-glycoprotein (P-gp) expression was detected by Western blot analysis. P-gp expression on the cell surface was observed by immunofluorescence. In-tracellular drug concentration was detected by flow cytometry. Drug resistance was detected by MTT assay. Results:The KG1a/SFRP5 cell, a KG1a cell expressing SFRP5, and the KG1a/eGFP cell, a KG1a cell expressing eGFP, were successfully constructed. The multidrug resistance protein 1 (mdr1) mRNA level in KG1a/SFRP5 cells was significantly decreased (P<0.01). P-gp expression in KG1a/SFRP5 cells was also downregulated. Fluorescence intensity of P-gp on the KG1a/SFRP5 cell surface was reduced. Rhodamine concentration in KG1a/SFRP5 cell was significantly increased (P<0.01). IC50 of KG1a/SFRP5 cell to Adriamycin was decreased as determined by the MTT method. Conclusion:SFRP5 expression in KG1a cells can downregulate MDR1 transcription and P-gp expression, increase intracellular drug concentration, and reverse multidrug resistance.