再生障碍性贫血患者血清对脐血CD34^+细胞cyclin D关键亚型表达的影响  

Effects of aplastic anemia patient serum on the expression of the crucial cyclin D isoform in cord blood CD34^+ cells

作　　者：孟凡凯[1] 谭细友[1] 孙汉英[1] 刘文励[1] 周剑锋[1] 刘丹[1] 付丽[1] 罗琳[1] 孙岚[2] 

机构地区：[1]华中科技大学同济医学院附属同济医院血液内科,湖北武汉430030 [2]温州医学院附属第一医院血液科,浙江温州325027

出　　处：《中国病理生理杂志》2006年第7期1362-1365,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30070326)

摘　　要：目的:确定控制脐血造血干/祖细胞增殖的细胞周期蛋白D(cyclinD)关键亚型,体外观察再生障碍性贫血(再障)患者血清对脐血造血干/祖细胞cyclinD关键亚型表达的影响,以探讨再障的发病机制。方法:用免疫磁珠阳性分选的方法分离脐带血CD34+细胞,用RT-PCR和Westernblotting方法检测其cyclinD各亚型的表达以筛选其关键亚型;用甲基纤维素半固体培养方法,观察再障血清对脐血CD34+细胞粒单系-集落形成单位(CFU-GM)形成的影响;再用半定量RT-PCR和Westernblotting方法检测再障血清对脐血CD34+细胞cyclinD关键亚型表达的影响。结果:RT-PCR和Westernblotting结果显示:控制脐血CD34+细胞增殖的cyclinD关键亚型为cyclinD3;再障血清可以显著抑制脐血CFU-GM的形成,并分别可以从mRNA和蛋白质水平抑制cyclinD3亚型的表达。结论:CyclinD3是控制脐血CD34+细胞增殖的关键cyclinD亚型,再障血清可以抑制其表达,这可能是再障造血抑制的一种机制。AIM ： To explore the pathogenesis of aplastic anemia （ AA）, we identified the crucial isoform of cyclin D that determine the proliferation of the cord blood CD34^＋ cells and observed effects of AA serum on the expression of crucial cyclin D isoform in umbilical cord blood CD34^＋ cells. METHODS： The CD34^＋ cells were isolated with MIDI - MACS system. The isoforms of cyclin D were detected by RT - PCR and Western blotting. Methylcellulose culture system was used to measure the formation of CFU - GM. The expression level of crucial cyclin D isoform was assayed by RT - PCR and Western blotting after the CD34^＋ cells were incubated in AA serum. RESULTS： The crucial cyclin D isoform in CD34^＋ cells was cyclin D3. The AA serum inhibited the formation of CFU - GM and down - regulated expression level of the cyclin D3 from the mRNA to protein level, respectively. CONCLUSION： The AA serum inhibits the proliferation of CD34^＋ cells and down- regulates level of cyclin D3, this may be one of hematopoiesis inhibition mechanisms in AA.

关 键 词：贫血 再生障碍性 干细胞 细胞周期蛋白D 

分 类 号：R331.2[医药卫生—人体生理学]