解剖学报
解剖學報
해부학보
ACTA ANATOMICA SINICA
2009年
6期
897-901
,共5页
宋姝丹%华自森%王建伟%王亚平
宋姝丹%華自森%王建偉%王亞平
송주단%화자삼%왕건위%왕아평
当归多糖%JAK_2%转录激活因子5%信号转导%免疫印迹法%K562细胞
噹歸多糖%JAK_2%轉錄激活因子5%信號轉導%免疫印跡法%K562細胞
당귀다당%JAK_2%전록격활인자5%신호전도%면역인적법%K562세포
Angelica polysaccharide%JAK_2%Activator of transcription 5%Signal transduction%Western blotting%K562 cell
目的 探讨当归多糖(APS)诱导K562细胞向红系样细胞分化相关的信号转导通路. 方法 实验分组:对照组采用常规方法培养;APS组在常规培养基础上加入APS(终浓度100～500mg/L),分别培养12h、1d、2d、3d.联苯胺染色与分光光度法检测经APS诱导后K562细胞向红系样细胞分化的血红蛋白表达;激光扫描共焦显微镜观察JAK_2、信号转导和转录激活因子5(STAT_5)在各组细胞内的分布;Western blotting检测细胞核、质蛋白中JAK_2、STAT_5的表达变化;免疫沉淀法检测质蛋白中JAK_2的磷酸化改变. 结果 经APS诱导后K562细胞的联苯胺染色阳性率增高,随着APS诱导浓度增加K562细胞合成血红蛋白也增加;APS诱导K562细胞12h、24h和48h,核蛋白中STAT_5表达较对照组增加,质蛋白中STAT_5表达减少;APS组与对照组K562细胞的JAK_2表达未见显著差异,但APS组的JAK_2磷酸化强于对照组. 结论 APS可诱导K562细胞向红系样细胞分化,其机制可能与APS启动JAK_2的酪氨酸磷酸化,继而引起STAT_5核转位,通过信号转导通路的调控影响细胞的增殖分化.
目的 探討噹歸多糖(APS)誘導K562細胞嚮紅繫樣細胞分化相關的信號轉導通路. 方法 實驗分組:對照組採用常規方法培養;APS組在常規培養基礎上加入APS(終濃度100～500mg/L),分彆培養12h、1d、2d、3d.聯苯胺染色與分光光度法檢測經APS誘導後K562細胞嚮紅繫樣細胞分化的血紅蛋白錶達;激光掃描共焦顯微鏡觀察JAK_2、信號轉導和轉錄激活因子5(STAT_5)在各組細胞內的分佈;Western blotting檢測細胞覈、質蛋白中JAK_2、STAT_5的錶達變化;免疫沉澱法檢測質蛋白中JAK_2的燐痠化改變. 結果 經APS誘導後K562細胞的聯苯胺染色暘性率增高,隨著APS誘導濃度增加K562細胞閤成血紅蛋白也增加;APS誘導K562細胞12h、24h和48h,覈蛋白中STAT_5錶達較對照組增加,質蛋白中STAT_5錶達減少;APS組與對照組K562細胞的JAK_2錶達未見顯著差異,但APS組的JAK_2燐痠化彊于對照組. 結論 APS可誘導K562細胞嚮紅繫樣細胞分化,其機製可能與APS啟動JAK_2的酪氨痠燐痠化,繼而引起STAT_5覈轉位,通過信號轉導通路的調控影響細胞的增殖分化.
목적 탐토당귀다당(APS)유도K562세포향홍계양세포분화상관적신호전도통로. 방법 실험분조:대조조채용상규방법배양;APS조재상규배양기출상가입APS(종농도100～500mg/L),분별배양12h、1d、2d、3d.련분알염색여분광광도법검측경APS유도후K562세포향홍계양세포분화적혈홍단백표체;격광소묘공초현미경관찰JAK_2、신호전도화전록격활인자5(STAT_5)재각조세포내적분포;Western blotting검측세포핵、질단백중JAK_2、STAT_5적표체변화;면역침정법검측질단백중JAK_2적린산화개변. 결과 경APS유도후K562세포적련분알염색양성솔증고,수착APS유도농도증가K562세포합성혈홍단백야증가;APS유도K562세포12h、24h화48h,핵단백중STAT_5표체교대조조증가,질단백중STAT_5표체감소;APS조여대조조K562세포적JAK_2표체미견현저차이,단APS조적JAK_2린산화강우대조조. 결론 APS가유도K562세포향홍계양세포분화,기궤제가능여APS계동JAK_2적락안산린산화,계이인기STAT_5핵전위,통과신호전도통로적조공영향세포적증식분화.
Objective To explore the relative signal transduction pathway of angelica polysaccharide(APS) inducing K562 cells toward erythroid differentiation. Methods K562 cells were divided into control group and ASP group, the control group cells were routinely cultured and APS group cells were treated with APS(final concentration is 100-500mg/L). By using of benzidine staining and spectrophotometry, the characteristics of erythroid differentiation of K562 cell induced by APS were detected;By using of laser confocal microscopy, the distribution of JAK_2 and STAT_5 in K562 cells was observed;By using of Western blotting, the expression of JAK_2 and STAT_5 in nucleus and cytoplasm of K562 cells was detected, By using of imm～uno～pre～cipi～ta～tion, the phosphorylation change of JAK_2 in cytoplasm was tested. Results After being induced by APS, the benzidine staining positive rate of K562 was increased.With increasing the concentration of APS, hemoglobin synthesis in K562 cells was promoted accordingly. After being cultured for 12 hours, 24 hours and 48 hour, the expression of STAT_5 in nucleus of K562 cells induced by APS was significantly higher than that of control group, however, expression of STAT_5 in cytoplasm of K562 cell induced by APS was obviously lower. The expression of JAK_2 in K562 cells was not different between APS group and control group, but the JAK_2 phosphorylation level of APS group was much higher than that of control group.Conclusion APS can induce erythroid differentiation of K562 cells, and the mechanisms may be that APS can promote the phosphorylationthe of JAK_2, then stimulating nuclear translocation of STAT_5.
