外源性FrzB拮抗Wnt信号转导并抑制人骨肉瘤细胞143B增殖  

Exogenous FrzB antagonizes the transduction of Wnt signaling and inhibits the proliferation of human osteosarcoma cell line 143B

作　　者：司维柯[1] 赵辰[2] 赵宸[1] 李军[1] 潘静[1] 李招权[1] 何通川[3] 

机构地区：[1]第三军医大学临床血液学教研室,重庆400038 [2]第三军医大学学员旅12队,重庆400038 [3]美国芝加哥大学医学中心分子肿瘤学研究室,芝加哥60637

出　　处：《免疫学杂志》2010年第3期202-205,共4页Immunological Journal

基　　金：美国癌症协会伊利诺斯州分会Concern-Brinson基金部分资助

摘　　要：目的 Wnt信号异常激活与肿瘤发生有关,观察Wnt信号的天然拮抗剂FrzB对Wnt信号转导的影响和对肿瘤细胞增殖抑制。方法采用AdEasy系统构建重组腺病毒AdFrzB;卡那霉素抗性筛选、酶切鉴定及荧光显微镜和Western blot检测标记基因GFP和His表达等方法鉴定AdFrzB。AdFrzB与AdWnt3A共感染人骨肉瘤细胞株143B,AdGFP与AdWnt3A共感染为对照,β-catenin/Tcf荧光素酶反应系统检测FrzB对Wnt信号转导的影响;MTT实验检测FrzB对细胞增殖的抑制。结果卡那霉素抗性筛选及酶切鉴定获得pAdFrzB-His;检测到报告基因GFP和His表达,确证AdFrzB构建成功。FrzB抑制了Wnt信号转导并抑制了143B细胞增殖。结论 AdEasy系统有效构建了AdFrzB;外源性FrzB拮抗Wnt信号转导并抑制143B细胞增殖,FrzB可能是一种天然抗肿瘤因子。Abnormal activation of Writ signaling pathway is associated with tumorigenesis. This study was to explore the effects of FrzB, a natural inhibitor of Wnt signaling, on the proliferation of tumor cells and the transduction of Wnt signaling. By using AdEasy protocol, recombinant adenovirns AdFrzB was constructed, and then identified by kanamycin selection and restriction enzymatic analysis, while tag genes were assayed with fluorescence microscope and Western blot analysis. MTT assay was performed to determine the FrzB＇s effects on the proliferation of human osteosarcoma cell line 143B which were infected with AdFrzB. The inhibitory effect of FrzB on Wnt signal was assessed with a β-catenin/Tcf reaction system in AdWnt3A and AdFrzB co-infected 143B cell line. AdFrzB was efficiently generated using the improved AdEasy system. We demonstrated that Exogenous FrzB could antagonize the transduction of Wnt signaling and inhibit 143B cell proliferation. Thus FrzB may be a new natural anti-tumor agent.

关 键 词：sFRP3/FrzB sFRPs WNT信号 143B细胞株 重组腺病毒 

分 类 号：R738.1[医药卫生—肿瘤]