豆类凝集素FRIL通过调控细胞周期分子的表达维持造血干／祖细胞的静息  

FRIL maintains quiescence of hematopoietic stem cells through regulation of cell cycle related factors

作　　者：李锦[1] 谢小燕[1] 王冬梅[1] 裴雪涛[1] 

机构地区：[1]军事医学科学院输血研究所干细胞研究室,北京100850

出　　处：《中华血液学杂志》2007年第1期37-40,共4页Chinese Journal of Hematology

基　　金：国家重点基础研究发展规划项目（2001CB509906）;国家高技术“863”计划领域重大专项资助项目（2006AA02A100）：北京市科委资助项目（H020220010190,Z0005190043331）

摘　　要：目的从细胞周期角度探讨豆类凝集素FRIL（Flt3 receptor—interacting lectin）体外维持造血干／祖细胞静息的分子机制。方法以添加FRIL、Flt3配体（FL）和不添加因子的培养基分别培养脐血CD34^＋细胞，采用RT-PCR和Western blot法分别从mRNA和蛋白水平检测细胞周期相关分子的表达。结果新分离的CD34^＋细胞中未检测到G0／G1期相关Cyclin和CDK蛋白的表达，培养3d时FRIL组CyclinD3、CDK6蛋白相对表达量（分别为483±63、553±39）低于两个对照组（FL组：2437±52、3209±98；空白对照组：914±105、1497±55）；培养3d时FRIL组P27蛋白相对表达量（0．312±0．030）低于对照组（FL组：0．787±0．024；空白对照组：0．616±0．029），但表达较高的P53蛋白（FRIL组、FL组、空白对照组分别为4．476±0．159、0．581±0．099、2．167±0．114）。FRIL组细胞周期正向调节因子的mRNA相对表达量低于或相当于FL组和空白对照组。结论FRIL通过抑制参与造血细胞周期调控的CyclinD3和CDK6的表达，延迟了造血干／祖细胞的细胞周期，P27在FRIL抑制造血干／祖细胞分化中发挥了重要作用，P53也参与了FRIL对造血干／祖细胞的维持作用。Objective To explore the mechanism of Fit3 receptor-interacting lectin（FRIL） maintains quiescence of hematopoietic stem cells （HSCs） in vitro. Methods Cord blood CD34^＋ cells were cultured in suspension medium supplemented with or without FRIL and FL. Ceils were collected at different time points and the expression of some cell cycle regulators, especially those involved in G0/G1 phase regulation were detected on mRNA and protein level. Results The expressions of G0/G1 phase related cyclins or CDKs were undetectable in the newly isolated CD34^ ＋ cells, expressions of CyclinD3, CDK6 and P27 were the lowest in FRIL cultured group after 3d＇ s culture （ FRIL group ： 483 ± 63,553 ± 39, 0.312 ± 0.030 ; FL group ： 2437 ± 52, 3209 ＋ 98, 0.787 ± 0.024 ; BLANK ： 914± 105, 1497 ± 55,0.616 ± 0. 029, respectively）, but the expression of 1）53 was the highest in FRIL group （ FRIL group ： 4. 476 ± 0. 159 ; FL group ： 0. 581 ± 0. 099, BLANK ： 2. 167±0.114 ）. The expression of positive regulators of cell cycle in FRIL group were the same as that of FL group and blank group or lower. Conclusion FRIL preserves HSCs effectively in vitro through the mechanisms of down-regulation of cyclinD3 and CDK6 and activation of P53. P27 is mostly involved in the differentiation of HSCs.

关 键 词：植物凝集素类 细胞周期 造血干细胞 

分 类 号：R686[医药卫生—骨科学]