期刊文献+

Role of Na^+/H^+ exchanger isoform-1 in doxorubicin-induced multidrug-resistance HL-60 cell line

Role of Na^+/H^+ exchanger isoform-1 in doxorubicin-induced multidrug-resistance HL-60 cell line
下载PDF
导出
摘要 Objective: To explore the roles of intracellular pH value (pHi) and sodium-hydrogen exchanger isoform-1 (NHE-1) in the mechanism of multidrug resistance of leukemia cells. Methods: Multidrug resistant cell line HL-60 induced by doxorubicin(DOX) (called as HL-60/DOX cells) and their parent cell line HL-60 were employed as experiment group and control group. The proliferation and chemosensitivity of the cells were studied by MTT assay, and the expression of multidrug resistance protein (MRP) was detected by immol/Lunocytochemistry. Meanwhile, pHi was measured by spectrofluorometery with a fluorescence dye BCECF-AM. Based on the pHi recovery curve after intracellular acid loading, the activity of NHE-1 was analyzed. The expression of NHE-1 mRNA and MRP mRNA were determined by semi-quantitative RT-PCR. Cell apoptosis was observed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), and apoptotic DNA was extracted and electrophoresed. Results: ① The IC 50 values for DOX, MTZ, VCR and homoharringtonine(HT), in HL-60/DOX cells were significantly higher than those in HL-60 cells (P<0.01). HL-60/DOX cells expressed abundant MRP, but HL-60 cells did not. ② pHi of HL-60/DOX cells were significantly higher than that of HL-60 cells(P<0.001). The expression and activity of NHE-1 in HL-60/DOX cells were significantly stronger than those of HL-60 cells. ③After administration of the specific NHE-1 inhibitor dimethyl amiloride (DMA) at a certain range of concentrations, compared with HL-60 cells, the rate of growth inhibition of HL-60/DOX cells increased significantly (P<0.05), the drug-sensitivity of HL-60/DOX cells was significantly sensitive (P<0.01), the expression of MRP and MRP mRNA decreased significantly (P<0.01), the apoptosis rate increased significantly (P<0.01). Conclusion: NHE-1 is involved in the drug-resistant mechanisms of multidrug-resistant HL-60 cells induced by DOX. The specific NHE-1 inhibitor DMA can partly reverse the multidrug resistance of HL-60 cells induced by DOX. Objective: To explore the roles of intracellular pH value (pHi) and sodium-hydrogen exchanger isoform-1 (NHE-1) in the mechanism of multidrug resistance of leukemia cells. Methods: Multidrug resistant cell line HL-60 induced by doxorubicin(DOX) (called as HL-60/DOX cells) and their parent cell line HL-60 were employed as experiment group and control group. The proliferation and chemosensitivity of the cells were studied by MTT assay, and the expression of multidrug resistance protein (MRP) was detected by immol/Lunocytochemistry. Meanwhile, pHi was measured by spectrofluorometery with a fluorescence dye BCECF-AM. Based on the pHi recovery curve after intracellular acid loading, the activity of NHE-1 was analyzed. The expression of NHE-1 mRNA and MRP mRNA were determined by semi-quantitative RT-PCR. Cell apoptosis was observed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), and apoptotic DNA was extracted and electrophoresed. Results: ① The IC 50 values for DOX, MTZ, VCR and homoharringtonine(HT), in HL-60/DOX cells were significantly higher than those in HL-60 cells (P<0.01). HL-60/DOX cells expressed abundant MRP, but HL-60 cells did not. ② pHi of HL-60/DOX cells were significantly higher than that of HL-60 cells(P<0.001). The expression and activity of NHE-1 in HL-60/DOX cells were significantly stronger than those of HL-60 cells. ③After administration of the specific NHE-1 inhibitor dimethyl amiloride (DMA) at a certain range of concentrations, compared with HL-60 cells, the rate of growth inhibition of HL-60/DOX cells increased significantly (P<0.05), the drug-sensitivity of HL-60/DOX cells was significantly sensitive (P<0.01), the expression of MRP and MRP mRNA decreased significantly (P<0.01), the apoptosis rate increased significantly (P<0.01). Conclusion: NHE-1 is involved in the drug-resistant mechanisms of multidrug-resistant HL-60 cells induced by DOX. The specific NHE-1 inhibitor DMA can partly reverse the multidrug resistance of HL-60 cells induced by DOX.
出处 《Journal of Medical Colleges of PLA(China)》 CAS 2004年第2期86-93,共8页 中国人民解放军军医大学学报(英文版)
基金 theResearchProjectSponsoredbyXinqiaoHospitalofThirdMilitaryMedicalUniversity (2 0 0 2 )
关键词 MULTIDRUG-RESISTANCE leukemia cells sodium-hydrogen exchanger isoform-1 intracellular pH value REVERSION 对碘氧基苯甲醚-1 阿霉素 多药耐药 HL-60细胞 离子交换 白血病
  • 相关文献

参考文献10

  • 1Come MG,Skladanowski AK,Laureng G et al.Dual mechanism of doxorubicin-induced cell death in both sensitive and MDR-resistant HL-60 cells[].British Journal of Cancer.1999
  • 2Weisburg JH,Roepe PD,Dzekunov S et al.Intracellular pH and multidrug resistance regulate complement-mediated cytotoxicity of nucleated human cells[].Journal of Biological Chemistry.1999
  • 3Garnovskaya MN,Mukhin YV,Turner JH et al.Mitogen-induced activation of Na+ H+ exchange in vascular smooth muscle cells involves janus kinase 2 and Ca2+ calmodulin[].Biochemistry.2003
  • 4Sergeant JM,Jailor CG.Appraisail of the MTT assay as a rapid test of chemo-sensitivity in acute myeloid leukemia[].British Journal of Cancer.1989
  • 5Ross DD,Novel.Mechanisms of drug resistance in leukemia[].Leukemia.2000
  • 6Come MG,Skladanowski AK,Laureng G et al.Dual mechanism of daunorubicin-induced cell death in both sensitive and MDR-resistant HL-60 cells[].British Journal of Cancer.1999
  • 7Batrakova EV,Li S,Elmquist WF et al.Mechanism of sensitization of MDR cancer cells by Pluronic block copolymers: Selective energy depletion[].British Journal of Cancer.2001
  • 8Wada H,Saikawa Y,Niida Y et al.Selectively induced high MRP gene expression in multidrug-resistant human HL60 leukemia cells[].Experimental Hematology.1999
  • 9Wahl ML,Owen JA,Burd R et al.Regulation of intracellular pH in human melanoma: potential therapeutic implications[].Molecular Cancer Therapeutics.2002
  • 10Quinn DA,Dahlberg CGW,Bonventre JP et al.The role of Na+ H+exchange and growth factors in pulmonary artery smooth muscle cell proliferation[].Am J Respir Cell Mol Biol.1996

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部