LiCl对脐带血有核细胞增殖作用和信号传导研究  

Effect of lithium chloride on umbilical cord blood nucleated cells in cell proliferation and signal pathway

作　　者：冯乐平[1] 陈再明 孙情[1] 于冉冉[1] 乔伟[1] 

机构地区：[1]桂林医学院生物技术学院,广西桂林541004 [2]广西灵川县医院,广西灵川541213

出　　处：《中国现代医学杂志》2011年第35期4371-4374,共4页China Journal of Modern Medicine

基　　金：国家自然科学基金(No:30860116);广西科技厅资助项目(No:0249020);广西科技厅科研基金(No:0728230);广西教育厅科研基金(院科字[2008]17号)

摘　　要：目的探讨不同浓度LiCl对脐带血有核细胞增殖、集落形成以及细胞凋亡的影响,观察在此过程中LiCl对脐带血有核细胞β-catenin表达的作用。方法应用红细胞裂解方法获得脐带血有核细胞,以5 000个/孔细胞接种到96孔板和1×105个/孔细胞接种到24孔板,应用不同浓度（10、20和40 mM）LiCl处理细胞,用MTT方法测定脐带血有核细胞增殖,以1×105个/孔细胞接种24孔细胞培养板,观察LiCl对细胞集落形成的影响;应用Hoechst 33342试剂盒荧光显微镜下检测细胞凋亡和应用Western blotting方法检测脐带血有核细胞β-catenin基因表达情况。结果随着LiCl浓度的不断增加（0～40 mM）和培养时间的不断延长（0~72h）,脐带血有核细胞增殖能力与相应对照组相比不断增加,分别呈现出不同程度的显著性差异（分别P〈0.05和P〈0.01）;应用20 mM浓度的LiCl培养10 d后,发现脐带血有核细胞集落形成能力与对照组相比差异有统计学意义（P〈0.05）,并呈现出明显的抑制细胞凋亡的作用;Western blotting结果发现,LiCl具有明显地诱导β-catenin基因表达的作用。结论 LiCl在一定条件下能够通过激活脐带血有核细胞内Wnt/β-catenin信号传导途径刺激细胞增殖,达到扩增脐带血有核细胞的目的。[Objective] To study the mechanism of umbilical cord blood nucleated cells proliferation, colony formation, cell apoptosis and cell signaling pathway treated with different concentrations of LiCl. [Methods] Collecting umbilical cord blood nucleated ceils applied by red blood cell lysis buffer, the cells were dividied into 96 well with 5 000 cells per well and 24 well plates with 1×10^5 cells per well, the cell treated with LiCl in 10 mM, 20 mM and 40 mM concentration respectively, then cell proliferation were measured with conventional MTT methods, the apoptotic cells were checked with Hoechst 33342 kit under flures- cence microscope and the cell colony-forming were observed. 13-catenin gene expression were deteehed with Western blot assay. [Results] With increasing LiC1 concentration and extending cell culture time, the cord blood nucleated cells proliferation increased （both of 20 mM and 40 mM LiCl concentration groups showed varying significantly differences respectively （P〈0.05 and P〈0.01）. After 10 days treated with the same LiCl concentration as above, cord blood nucleated cell colony-forming were far more than those of control group that were showed significantly difference （P 〈0.05）, but the cell apoptosis naturely inhibition was obviously showed. Western blot results showed that LiC1 significantly induced [3-catenin gene expression when the LiC1 concentration was in 20 raM. [ Conclusion] Under certain concentration, LiC1 can stimulate the cord blood nu- cleated cells proliferation by activating Wnt/β-catenin signal transduction pathway.

关 键 词：LICL 脐带血 有核细胞 细胞增殖 细胞凋亡 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]