经典Wnt信号通路在非霍奇金淋巴瘤细胞株中作用的初步研究  

Preliminary study of the role of canonical Wnt signalling pathway in non-Hodgkin’s lymphoma cell lines

作　　者：曲琦[1] 黄洪晖[1] 陈芳源[1] 钟华[1] 王婷[1] 钟济华[1] 

机构地区：[1]上海交通大学医学院附属仁济医院血液科,上海200127

出　　处：《肿瘤》2013年第2期144-149,共6页Tumor

基　　金：国家自然科学基金面上项目资助课题(编号:81172253);上海交通大学医学院附属仁济医院重点学科资助项目(编号:RJ4101306)

摘　　要：目的:通过检测经典Wnt信号通路在非霍奇金淋巴瘤(non-Hodgkin’s lymphoma,NHL)细胞株中是否存在异常激活,以探讨其与NHL发病的相关性。方法:选取人NHL细胞株SUDHL-4、Raji和Namalwa(以正常人淋巴细胞作为对照),采用蛋白质印迹法检测各细胞株中β-链蛋白(β-catenin)、糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)及其失活形式Ser9被磷酸化的GSK-3β[phospho-glycogen synthase kinase-3β(Ser9),p-GSK-3β(Ser9)]蛋白的表达情况;应用实时荧光定量-PCR技术检测β-catenin编码基因CTNNB1、经典Wnt通路上游相关调控因子低密度脂蛋白受体相关蛋白5(low density lipoprotein receptor related protein5,LRP5)基因及靶基因c-Myc的转录表达水平。结果:与正常对照组淋巴细胞相比较,3株NHL细胞中总蛋白及细胞核内β-catenin蛋白的表达水平明显上调;p-GSK-3β(Ser9)蛋白较正常对照也存在不同水平表达上调的现象,而GSK-3β蛋白的表达在各组间差异无统计学意义。NHL细胞中CTNNB1、LRP5及c-MycmRNA表达水平与正常对照相比,均有明显提高。结论:β-catenin作为经典Wnt信号通路重要的激活标志,其在NHL中的高表达及经典Wnt通路其他相关分子异常,说明经典Wnt信号通路可能是导致NHL的分子作用机制之一。Objective： To determine whether the canonical Wnt signalling pathway is abnormally activated in NHL （non-Hodgkin＇s lymphoma）, and to explore its association with pathogenesis of NHL. Methods： The expression levels of β-catenin, GSK-3β （glycogen synthase kinase-3β） and its inactive form p-GSK-3β （Ser9） （phosphorylation of GSK-3β at Ser9 residue） in NHL cell lines including SUDHL-4, Raji and Namalwa were detected by Western blotting, and the normal human lymphocytes were served as the controls. The mRNA levels of CTNNB1 gene （encoding β-catenin）, canonical Wnt signaling pathway- relevant gene LRP5 （low-density lipoprotein receptor-related protein 5） and target gene c-Myc were detected by real-time fluorogenic quantitative PCR. Results： The expressions of total cellular proteins and β-catenin protein in cell nuclei were significantly up-regulated in three NHL cell lines as compared with in those normal human lymphocytes. The elevated expression levels of p-GSK-3β （Ser9） were observed in lymphoma cell lines, but the expression level of GSK-3β was not significantly different between the lymphoma cell lines and the normal human lymphocytes. The mRNA expression levels of CTNNB1, LRP5 and c-Myc were significantly higher in lymphoma cell lines than those in the normal human lymphocytes. Conclusion： Given the important role of β-catenin as an activation marker in canonical Wnt signaling pathway, the evidence of up-regulated expression of β-catenin and the changes of other relevant molecules in canonical Wnt signaling pathway suggests that the constitutional activation of canonical Wnt signaling pathway may contribute to the pathogenesis of NHL.

关 键 词：淋巴瘤 非霍奇金 WNT信号通路 Β-CATENIN 糖原合成酶激酶3 

分 类 号：R733.1[医药卫生—肿瘤]