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.docVIP

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 胡艳妮，徐酉华，郑改焕，罗 庆，沈亚莉 (400014 重庆，重庆医科大学附属儿童医院血液科) [摘要] 目的? 应用重组腺病毒介导的Wnt-3a激活急性T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-3a mRNA的表达；Western blot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Cyclin D1 mRNA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。Western blot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，与对照组相比差异有统计学意义（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Cyclin D1 mRNA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Cyclin D1的表达实现。 [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 [中图法分类号] [文献标志码] A Exogenous Wnt-3a promotes proliferation and cell cycle progression of T-lineage acute lymphoblastic leukemia cells Hu Yanni, Xu Youhua, Zheng Gaihuan, Luo Qing，Shen Yali (Department of Hematology, Affiliated Children’s Hospital, Chongqing Medical University, Chongqing 400014, China) [Abstract] 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 blot. Proliferation of Jurkat cells was detected by MTT assay. Progression of Jurkat ce