CAJ | 학술논문

目的研究经bcr-abl诱导形成的白血病细胞中β-catenin缺失对Stat-5α蛋白磷酸化水平的影响.方法利用已经建立的特异性骨髓造血系统β-catenin基因敲除小鼠,分离其骨髓细胞,应用逆病毒感染方法将外源性bcr-abl基因片段导入β-catenin缺失的骨髓细胞内,免疫荧光和Western blotting方法检测Stat-5 α蛋白磷酸化水平,应用real-time PCR及Western blotting分别检测β-catenin缺失细胞及对照细胞bcr-abl转录及蛋白表达.结果与对照组相比,β-catenin缺失的白血病细胞Stat-5α磷酸化水平明显降低,而Stat-5α总蛋白量变化不明显;Western blotting结果显示β-catenin缺失的慢性髓细胞白血病(CML)细胞中酪氨酸磷酸化总水平及bcr-abl蛋白表达明显降低,而β-catenin缺失的急性淋巴细胞白血病(ALL)细胞中酪氨酸磷酸化总水平及bcr-abl蛋白表达无明显改变.结论 bcr-abl诱导形成的白血病转化细胞中,β-catenin缺失抑制bcr-abl蛋白表达及Stat-5 α磷酸化.
목적 연구경bcr-abl유도형성적백혈병세포중β-catenin결실대Stat-5α단백린산화수평적영향.방법 이용이경건립적특이성골수조혈계통β-catenin기인고제소서,분리기골수세포,응용역병독감염방법장외원성bcr-abl기인편단도입β-catenin결실적골수세포내,면역형광화Western blotting방법검측Stat-5 α단백린산화수평,응용real-time PCR급Western blotting분별검측β-catenin결실세포급대조세포bcr-abl전록급단백표체.결과 여대조조상비,β-catenin결실적백혈병세포Stat-5α린산화수평명현강저,이Stat-5α총단백량변화불명현;Western blotting결과현시β-catenin결실적만성수세포백혈병(CML)세포중락안산린산화총수평급bcr-abl단백표체명현강저,이β-catenin결실적급성림파세포백혈병(ALL)세포중락안산린산화총수평급bcr-abl단백표체무명현개변.결론 bcr-abl유도형성적백혈병전화세포중,β-catenin결실억제bcr-abl단백표체급Stat-5 α린산화.
Objective To investigate the influence of β-catenin gene deletion on Stat-5α phosphorylation in bcr-abl induced leukemia cells. Methods The established conditonal hematopoitic β-catenin knockout mice were used to isolate bone marrow cells. Exogenous bcr-abl fusion gene was transduced to these bone marrow cells by retroviral infection with intent to transfom them to leukemia cells.Immunofluorescence was performed to detect the phosphorylation status of Stat-5α in both β-catenin deletion cells and control cells. bcr-abl transcription and protein levels were evaluated with real-time PCR and western blotting. Results Phosphorylation of Stat-5α was reduced significantly in β-catenin deletion leukemia cells on comparison with control cells despite that total Stat-5α protein showed no obvious changes. Total tyrosine phosphorylation and bcr-abl protein expression were reduced in bcr-abl induced β-catenin deletion CML cells,on the contrary, both of the reduction were not seen in bcr-abl induced β-catenin deletion ALL cells.Conclusion Loss of β-catenin inhibits both Stat-5α phosphorylationin and bcr-abl expression in bcr-abl induced leukemia cells.