电压门控性钾通道在急性髓系白血病细胞增殖及凋亡中的作用  

Effects of voltage-gated potassium channel on cell cycle and apoptosis of AML

作　　者：张秋[1] 王巍[1] 王连杰[1] 何婉婷[1] 任雨悦 

机构地区：[1]哈尔滨医科大学附属第二医院血液科,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2015年第2期99-102,共4页Journal of Harbin Medical University

基　　金：国家自然科学基金资助项目(81001051);黑龙江省青年基金(QC2010019);黑龙江省教育厅新世纪人才项目(1252-NCET-014)

摘　　要：目的研究电压门控性钾通道在急性髓系白血病(acute myelocytic leukemia,AML)细胞增殖及凋亡中的作用。方法通过台盼蓝拒染法检测AML细胞株THP-1生存活力;应用Annexin V/PI双标法和DAPI荧光染色法检测细胞凋亡;应用流式细胞术检测[Ca2+]i变化。结果电压门控性钾通道抑制剂4-氨基吡啶(4-aminopyrine,4-AP)作用72 h后,THP-1细胞生存活力明显下降,IC50为3.68 mmol/L。药物作用72 h后,DAPI荧光染色示4-AP 2 mmol/L组、4 mmol/L组细胞凋亡率分别为(14.38±1.82)%、(32.91±3.47)%,与对照组(6.35±0.29)%比较差异显著(P<0.05);Annexin V/PI双标记法示4-AP 2 mmol/L组、4 mmol/L组细胞凋亡率分别为(16.89±2.49)%、(38.26±4.03)%,与对照组(6.06±0.32)%比较差异均显著(P<0.05)。2及4 mmol/L 4-AP作用1 min后,THP-1细胞内Fluo-3/AM荧光强度分别为(378.36±10.84)、(391.31±24.78),与对照组(57.24±7.18)相比具有显著性差异(P<0.05)。结论电压门控性钾通道阻滞剂4-AP抑制AML细胞生存活力并可诱导其凋亡,提示电压门控性钾通道可能是AML治疗的潜在靶点。Objective To investigate the effect of voltage-gated potassium channel on the cell cycle and apoptosis of acute myeloid leukemia （ AML）.Methods The effects of AML cell lines THP-1 on cells survival were determined by Trypan blue exclusion .The apoptosis rate was analyzed by using Annexin V-FITC double labeling and DAPI assay .The level of intracellular free Ca2＋was observed by flow cytometry .Resul ts THP1-cell viability was significantly de-creased with 4-aminopyridine treatment for 72 h, IC50 was 3.68 mmol/L.After 72 h, DAPI staining showed that the apoptosis rate of THP-1 in 4-AP 2 mmol/L group and 4 mmol/L group were （14.38 ±1.82）%and （32.91 ±3.47）%respectively, and were all significantly larger＆nbsp;than that of the control group （6.35 ±0.29 ） , P〈0.05 .Annexin V-FITC/PI double staining showed that the apoptosis rate of THP-1 were （16.89 ±2.49）%and （38.26 ±4.03）%respec-tively, and were all significantly larger than that of the control group （6.06 ±0.32）, P〈0.05. After 1 min, the Fluo-3/AM fluorescence intensity of THP-1 cell lines in 4-AP 2 mmol/L group and 4 mmol/L group were （378.36 ±10.84 ） and （391.31 ±24.78 ） respectively,and were all significantly larger than that of the control group （57.24 ±7.18 ） , P〈0.05 .Conclusion 4-AP could decrease the ability of cell growth and apoptosis of AML cells by blocking the voltage-gated potassium channel , which provides new ideas in the targeted therapy of AML .

关 键 词：急性髓系白血病 电压门控性钾通道 4-氨基吡啶 增殖 凋亡 钙离子 

分 类 号：R733.71[医药卫生—肿瘤]