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《Acta Academiae Medicinae Militaris Tertiae》 2007-04
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Reversal of multi-drug resistance in HL-60/ADM cells by dimethyl amiloride in vitro

CHANG Cheng,KONG Pei-yan,CHEN Xing-hua,PENG Xian-gui,LIU Lin,LIU Hong,ZENG Dong-feng,LIANG Xue,WANG Qing-yu(Department of Hematology,Xinqiao Hospital,Third Military Medical University,Chongqing 400037,China)  
Objective To investigate the reversal and the possible mechanism of multidrug-resistance(MDR)by dimethyl amiloride(DMA),a specific inhibitor of Na +/H +exchanger-1(NHE-1),in HL-60/ADM cell line in vitro.Methods After DMA treatment,the chemosensitivity of HL-60/ADM cells was studied by MTT assay.Intracellular concentration of daunorubicin(DNR)was determined by flow cytometry.Expression of mrp mRNA,bcl-2 mRNA,bax mRNA was assayed by semi-quantitive RT-PCR.MRP protein expression was detected immunohistochemically.Cell apoptosis was observed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL).Results After treated with 50 μmol/L DMA for 24 h,IC 50 of HL-60/ADM cells to ADM,MIT,VCR and HAR decreased from(839.5±45.2),(321.2±28.9),(70.5±15.5),(65.2±20.7)ng/ml to(180.5±38.9),(76.4±12.0),(30.2±7.5),(31.7±7.2)ng/ml.After treated with 50 μmol/L or 100 μmol/L DMA for 24 h,the intracellular concentration of DNR in HL-60/ADM cells increased from 33.56 to 48.74 or 70.10,and expression of mrp mRNA,MRP protein and bcl-2 mRNA decreased significantly,but bax mRNA increased significantly.After treated with 100 μmol/L DMA for 24 h,the rate of apoptotic cells in HL-60/ADM cells treated with 5 μg/ml ADM increased significantly.Conclusion DMA could reverse MDR of HL-60/ADM cells in vitro,and the mechanisms were associated with decrease of MRP expression and induction of cell apoptosis through bcl-2/bax pathway.
【Fund】: 第三军医大学新桥医院创新性基金(2001D068)~~
【CateGory Index】: R733.7
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