外源性Wnt-3a促进急性T淋巴细胞白血病细胞增殖与细胞周期进展  

Exogenous Wnt-3a promotes proliferation and cell cycle progression of T-lineage acute lymphoblastic leukemia cells

作　　者：胡艳妮[1] 徐酉华[1] 郑改焕[1] 罗庆[1] 沈亚莉[1] 

机构地区：[1]重庆医科大学附属儿童医院血液科,重庆400014

出　　处：《第三军医大学学报》2010年第23期2511-2514,共4页Journal of Third Military Medical University

摘　　要：目的应用重组腺病毒介导的Wnt-3a激活急性T淋巴细胞白血病细胞株Jurkat中的经典Wnt/β-catenin信号途径,观察该途径激活后对Jurkat细胞增殖及细胞周期的影响,初步探讨其在急性T淋巴细胞白血病分子发病机制中的作用。方法将携带Wnt-3a基因的重组腺病毒(Ad5-Wnt-3a)感染Jurkat细胞,实验分为3组:实验组(Jurkat/Ad5-Wnt-3a)、空载体组(Jurkat/Ad5-GFP)和空白对照组(Jurkat)。转染后,采用RT-PCR方法检测Jurkat细胞内Wnt-3amRNA的表达;Westernblot检测细胞内β-catenin蛋白的表达;MTT法检测细胞增殖,FCM法检测细胞周期,RT-PCR检测Wnt信号通路下游靶基因c-myc和cyclinD1mRNA表达。结果重组腺病毒Ad5-Wnt-3a转染Jurkat细胞后48h,实验组有效表达Wnt-3a的基因。Westernblot结果显示,与对照组比较,实验组细胞内β-catenin蛋白表达明显增加(P<0.05)。MTT结果显示,Wnt-3a转染后48h和72h可明显促进Jurkat细胞的增殖,与对照组相比差异有统计学意义(P<0.05)。FCM检测发现,Wnt-3a转染后可引起细胞G0/G1期下降,S期升高(P<0.05)。RT-PCR检测结果显示细胞内c-myc和cyclinD1mRNA表达升高,与对照组相比差异有统计学意义(P<0.05)。结论腺病毒介导的Wnt-3a可以激活Jurkat细胞中的经典Wnt/β-catenin信号途径,使Jurkat细胞增殖,促进细胞周期进展,其可能的机制是通过调控其下游靶基因c-myc和cyclinD1的表达实现。Objective To study the canonical Wnt/β-catenin signaling of activated Wnt-3a T-lineage acute lymphocytic leukemia cell line Jurkat with recombinant adenovirus vector, observe its effect on proliferation and cell cycle progression of Jurkat cells, and investigate its cellular mechanism underlying acute lymphocytic leukemia. Methods Jurkat cells, infected with recombinant adenovirus carrying the Wnt-3a gene were divided into experimental group （Jurkat/Ad5-Wnt-3a）, empty vector group （Jurkat/Ad5-GFP）, and blank control group （Jurkat）. Wnt-3a mRNA expression in Jurkat cells was detected by RT-PCR. β-catenin protein level was measured by Western blotting. Proliferation of Jurkat cells was detected by MTT assay. Progression of Jurkat cell cycle was detected by flow cytometry （FCM）. c-myc and cyclin D1 mRNA expression was detected by RT-PCR. Results Wnt-3a was effectively expressed in Jurkat cells 48 h after transfection with recombinant adenovirus vector. Western blotting showed that the β-catenin protein level was significantly higher in experimental group than in blank control group（P0.05）. MTT assay showed that the proliferation level of Jurkat cells was markedly higher in experimental group than in blank control group 48 and 72 h after transfection with Ad5-Wnt-3a（P0.05）. FCM showed that the number of Jurkat cells at G0/ G1 phase was decreased and those at S phase was increased. RT-PCR showed that c-myc and cyclin D1 mRNA expressions were significantly higher in experimental group than in blank control group（P0.05）. Conclusion Recombinant adenovirus vector-mediated Wnt-3a activates the canonical Wnt/β-catenin signaling in Jurkat cells and promotes their proliferation and cell cycle progression by down-regulating the expressions of c-myc and cyclin D1.

关 键 词：经典Wnt/β-catenin信号途径 急性T淋巴细胞白血病 细胞增殖 细胞周期 

分 类 号：R394.3[医药卫生—医学遗传学] R730.23[医药卫生—基础医学]