哈尔滨医科大学学报
哈爾濱醫科大學學報
합이빈의과대학학보
JOURNAL OF HARBIN MEDICAL UNIVERSITY
2015年
2期
99-102
,共4页
张秋%王巍%王连杰%何婉婷%任雨悦
張鞦%王巍%王連傑%何婉婷%任雨悅
장추%왕외%왕련걸%하완정%임우열
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
急性髓繫白血病%電壓門控性鉀通道%4-氨基吡啶%增殖%凋亡%鈣離子
급성수계백혈병%전압문공성갑통도%4-안기필정%증식%조망%개리자
acute myeloid leukemia%voltage-gated potassium channel%4-aminopyridine%pro-liferation%apoptosis%calcium
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。
目的：研究電壓門控性鉀通道在急性髓繫白血病（ acute myelocytic leukemia ，AML）細胞增殖及凋亡中的作用。方法通過檯盼藍拒染法檢測AML細胞株THP-1生存活力；應用Annexin V／PI雙標法和DAPI熒光染色法檢測細胞凋亡；應用流式細胞術檢測[ Ca2＋] i變化。結果電壓門控性鉀通道抑製劑4-氨基吡啶（4-aminopyrine，4-AP）作用72 h後，THP-1細胞生存活力明顯下降，IC50為3.68 mmol／L。藥物作用72 h 後， DAPI 熒光染色示4-AP 2 mmol／L 組、4 mmol／L 組細胞凋亡率分彆為（14.38±1.82）％、（32.91±3.47）％，與對照組（6.35±0.29）％比較差異顯著（P＜0.05）；Annexin V／PI雙標記法示4-AP 2 mmol／L組、4 mmol／L組細胞凋亡率分彆為（16.89±2.49）％、（38.26±4.03）％，與對照組（6.06±0.32）％比較差異均顯著（P＜0.05）。2及4 mmol／L 4-AP作用1 min後，THP-1細胞內Fluo-3／AM熒光彊度分彆為（378.36±10.84）、（391.31±24.78），與對照組（57.24±7.18）相比具有顯著性差異（ P＜0.05）。結論電壓門控性鉀通道阻滯劑4-AP抑製AML細胞生存活力併可誘導其凋亡，提示電壓門控性鉀通道可能是AML治療的潛在靶點。
목적：연구전압문공성갑통도재급성수계백혈병（ acute myelocytic leukemia ，AML）세포증식급조망중적작용。방법통과태반람거염법검측AML세포주THP-1생존활력；응용Annexin V／PI쌍표법화DAPI형광염색법검측세포조망；응용류식세포술검측[ Ca2＋] i변화。결과전압문공성갑통도억제제4-안기필정（4-aminopyrine，4-AP）작용72 h후，THP-1세포생존활력명현하강，IC50위3.68 mmol／L。약물작용72 h 후， DAPI 형광염색시4-AP 2 mmol／L 조、4 mmol／L 조세포조망솔분별위（14.38±1.82）％、（32.91±3.47）％，여대조조（6.35±0.29）％비교차이현저（P＜0.05）；Annexin V／PI쌍표기법시4-AP 2 mmol／L조、4 mmol／L조세포조망솔분별위（16.89±2.49）％、（38.26±4.03）％，여대조조（6.06±0.32）％비교차이균현저（P＜0.05）。2급4 mmol／L 4-AP작용1 min후，THP-1세포내Fluo-3／AM형광강도분별위（378.36±10.84）、（391.31±24.78），여대조조（57.24±7.18）상비구유현저성차이（ P＜0.05）。결론전압문공성갑통도조체제4-AP억제AML세포생존활력병가유도기조망，제시전압문공성갑통도가능시AML치료적잠재파점。
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 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 .
