水通道蛋白1基因过表达对K562细胞红系分化和增殖的影响  

Effects of aquaporin-1 gene overexpression on erythroid differentiation and proliferation of chronic myeloid leukemia K562 cells

作　　者：危敏[1] 姜立[2] 王妮莎[1] 马文丽[1] 

机构地区：[1]南方医科大学基因工程研究所,广州510515 [2]广东省人民医院科教处,广州510080

出　　处：《肿瘤》2012年第7期489-494,共6页Tumor

基　　金：国家自然科学基金资助项目(编号:30901721)

摘　　要：目的:探讨水通道蛋白1(aquaporin-1,AQP1)过表达对人慢性髓细胞白血病(chronic myeloid leukemia,CML)K562细胞红系分化和增殖的影响。方法:以人脑cDNA文库为模板,通过PCR扩增出AQP1基因的编码序列,构建pBABE-puro-AQP1真核表达载体;感染K562细胞,筛选建立稳定过表达AQP1基因的K562细胞株(命名为K562-AQP1);实时荧光定量PCR法、细胞免疫荧光染色法及蛋白质印迹法分别检测AQP1转录和蛋白表达水平。通过MTT法检测细胞生长增殖、实时荧光定量PCR法检测γ珠蛋白表达和分光光度法检测血红蛋白含量,研究AQP1过表达对K562细胞红系分化和增殖的影响。结果:与空载体对照组相比,pBABE-puro-AQP1转染入K562细胞后AQP1 mRNA和蛋白表达水平皆有显著升高(P<0.01),K562-AQP1细胞中红系分化指标γ珠蛋白和血红蛋白表达水平明显增加,同时细胞生长速度明显降低(P<0.05)。结论:AQP1过表达可以显著促进K562细胞向红系分化,同时抑制细胞增殖。推测AQP1可能成为临床诱导分化治疗CML的基因靶点之一。Objective： To explore the effects of aquaporin-1 （AQP1） overexpression on erythroid differentiation and cell proliferation of human chronic myeloid leukemia K562 cells in vitro. Methods： The coding sequence ofAQP1 gene was amplified by PCR from Human Brain cDNA Libraries. The eukaryotic expression vector pBABE-puro-AQP1 was constructed and transfected into K562 cells. The K562 cells stably overexpressing AQP1 protein （K562-AQP1 cells） were selected. Real-time PCR and the immunocytochemical staining and Western blotting were used to examine the transcription level of AQP1 mRNA and the expression of AQP1 protein, respectively. The cell proliferation was detected by MI-I- assay, the expression of gamma-globin was detected by real-time PCR, and the haemoglobin content was detected by ultraviolet spectrophotometry. Then the effects ofAQP1 overexpression on erythrod differentiation and proliferation of K562 cells were analyzed. Results： As compared with transfection with empty vector, the expression levels of AQP1 mRNA and protein in K562 cells transfected with pBABE-puro- AQP1 were both significantly increased （P〈0.01）. The contents of gamma-globin and haemoglobin relating to erythroid differentiation were significantly increased in K562-AQP1 cells, while the cell proliferation rate was markedly reduced （P〈0.05）. Conclusion： The overexpression of AQP1 gene can obviously promote the erythroid differentiation of K562 cells and inhibit the cell proliferation. It is suggested that AQP1 may become a gene target of differentiation induction therapy for chronic myeloid leukemia.

关 键 词：白血病 髓样 水通道蛋白1 基因表达 细胞分化 细胞增殖 K562细胞 

分 类 号：R733.72[医药卫生—肿瘤]