蛋白激酶C亚型在小鼠胚胎干细胞中的表达研究  

Study on the Expressions of Protein Kinase C Isoenzyme in EmbryonicStem Cells of Mouse

作　　者：高前应[1] 葛坚[1] 王智崇[1] 杨智宽[1] 黄丹平[1] 袁钊辉[1] 陈慧怡[1] 陶靖[1] 

机构地区：[1]中山大学中山眼科中心,广东广州510060

出　　处：《中山大学学报（医学科学版）》2003年第4期312-315,F002,共5页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家重点基础研究发展规划基金(973)子项目(G1999054301);国家自然科学基金(30171001);广东省重点科技攻关项目(A302020101);广东省自然科学基金(990093)

摘　　要：[目的]研究5种蛋白激酶C(PKC)亚型PKC_α、PKC_β、PKC_γ、PKC_δ和PKC_ε在小鼠胚胎干细胞(ES细胞)株ES-BALB/c的表达情况。[方法]ES-BALB/c细胞株用普通ES细胞培养液培养,分别种植于预置有盖玻片的6孔板和直径100mm的培养皿,培养2d和4d后分别进行PKC亚型免疫组织化学和Western印迹检测。[结果]免疫组化发现PKC_α在ES-BALB/c细胞有广泛的棕黄色的阳性表达,以细胞浆和细胞核最甚,而PKC_β、PKC_γ、PKC_δ和PKC_ε4种亚型无明显表达;Western印迹发现PKC_α出现一条蛋白印迹条带,分子质量约为84ku,而PKC_β、PKC_γ、PKC_δ和PKC_ε4种亚型无蛋白印迹条带。[结论]PKC_α在ES细胞阶段即有表达,在将来的细胞分化阶段可能有非常重要的作用,而PKC_β、PKC_γ、PKC_δ和PKC_ε4种亚型在ES细胞阶段没有表达。[Objective] To study the expressions of the protein kinase C(PKC) isoenzyme PKCα, PKCβ, PKCγ, PKCδ and PKCε in embryonic stem cells(ES cells) of mouse. [Methods] ES-BALB/c cells were incubated in the ES conditioned medium, and cultured in 6-well culture dishes with coated glass coverslip and in 100-mm dishes for 2 and 4 days, respectively. Immunohistochemistry and Western blot assay were performed to identify the PKC isoform expression. [Results] Staining of PKCα but PKCβ, PKCγ, PKCδ and PKCε was strong and distributed throughout the cells, especially in the cytoplasm and nucleus. Western blot analysis exhibited a band of approximately 84 ku with the antibody specific to PKCα. No reactions were shown for other four PKC isoenzyme, PKCβ, PKCγ, PKCδ and PKCε. [Conclusion] PKCα is expressed at the stage of ES cells and probably plays an very important role in differentiation , while PKCβ,PKCγ, PKCδ and PKCε are not expressed at the same time.

关 键 词：胚胎干细胞 蛋白激酶C 亚型 蛋白表达 免疫组织化学 WESTERN印迹法 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] Q51[医药卫生—基础医学]