骨髓间充质干细胞靶向p16^(INK4a)蛋白诱导慢性髓细胞性白血病细胞K562衰老的分子机制  

Moleecular Mechanism of K562 Senescence in Chronic Myeloigenous Leukemia Cells Induced by Targeting of P16^(INK4a) Protein by Bone Marrow Mesenchymal Stem Cells

作　　者：周雯 唐紫云 何思悦 刘小虎 周玥 Zhou Wen;Tang Ziyun;He Siyue;Liu Xiaohu;Zhou Yue(Pre-clinical College,Dali University,Dali,Yunnan 671000,China)

机构地区：[1]大理大学基础医学院,云南大理671000

出　　处：《大理大学学报》2022年第10期39-46,共8页Journal of Dali University

基　　金：国家自然科学基金项目(81860038,81660731)。

摘　　要：目的:探究小鼠骨髓间充质干细胞通过p16^(INK4a)蛋白诱导K562细胞衰老的作用机制。方法:从小鼠骨髓中分离、纯化间充质干细胞,使用流式细胞术鉴定表面标志物、检测K562细胞周期变化;油红O和茜素红染色鉴定其成脂、成骨分化能力;CCK-8法检测K562细胞增殖能力;SA-β-Gal染色检测K562细胞衰老情况;蛋白印迹法检测p16^(INK4a)蛋白的表达。结果:骨髓间充质干细胞呈成纤维样贴壁生长,低表达造血干细胞标志物CD45,高表达干细胞表面标志物CD29、CD44;具有成骨、成脂分化能力。骨髓间充质干细胞能够抑制K562细胞的增殖;阻滞K562细胞周期于G0/G1期;提高SA-β-Gal的活性和p16^(INK4a)蛋白的表达。结论:骨髓间充质干细胞能够通过上调p16^(INK4a)蛋白的水平诱导K562细胞衰老。Objective:To explore the mechanism of mouse bone marrow mesenchymal stem cell(BMMSC)inducing K562 cells aging through p16^(INK4a) protein.Methods:Mesenchymal stem cells were isolated and purified from mouse bone marrow.The changes of the K562 cell cycle were detected and the surface markers were identified by flow cytometry.The ability of adipogenic and osteogenic differentiation was identified by oil red O and alizarin red staining.The proliferation of K562 cells was detected by CCK-8 method.The senescence of K562 cells was detected by SA-β-Gal staining.The expression of p16^(INK4a) protein was detected by Western blot.Results:BMMSC showed fibroblast-like adherent growth,with low expression of hematopoietic stem cell marker CD45 and high expression of stem cell surface markers CD29 and CD44,which had osteogenic and adipogenic differentiation ability.BMMSC could inhibit the proliferation of K562 cells,block K562 cell cycle in G0/G1 phase,and improve SA-β-Gal activity and p16^(INK4a) protein expression.Conclusion:BMMSC can induce the senescence of K562 cells by up-regulating the level of p16^(INK4a) protein.

关 键 词：骨髓间充质干细胞 衰老 白血病 K562细胞 p16^(INK4a)蛋白 

分 类 号：R551.3[医药卫生—血液循环系统疾病]