p27^(kip1)及其出核相关分子激活蛋白1辅因子在淋巴瘤细胞中的表达及相互关系  

Expression and relationship of p27^(kipl)and it's nuclear export factor Jabl in Lymphoma cell Jurkat

作　　者：王燏婵[1] 赵玥铭[1] 沈爱国[1] 陆建新[1] 张冬梅[1] 何松[2] 程纯[1] 

机构地区：[1]南通大学微生物与免疫学教研室,226001 [2]南通大学附属肿瘤医院病理科

出　　处：《中华血液学杂志》2007年第12期813-817,共5页Chinese Journal of Hematology

基　　金：江苏省高校自然科学研究计划(04KJB320114);江苏省科技指导性计划(BS2004526);江苏省卫生科研项目(H200632)

摘　　要：目的探讨 p27^(kip1)及其出核相关分子激活蛋白1(AP-1)的辅助因子 Jab1在淋巴瘤细胞中的表达变化及相互关系。方法采用血清饥饿合并释放方法同步化处理淋巴瘤细胞系 Jurkat 和Raji 细胞,分别采用 Western blot 及 RT-PCR 技术检测 p27^(kip1)、Jab1在淋巴瘤细胞中的蛋白表达水平和mRNA 表达水平变化;采用来普霉素 B(LMB)刺激增殖过程中的淋巴瘤细胞,检测 p27^(kip1)、Jab1的表达变化;构建人 Jab1基因的 pcDNA3.1-myc-Jab1的真核表达质粒,脂质体转染 Jurkat 细胞,配合免疫荧光技术检测 p27^(kip1)的亚细胞定位情况;免疫沉淀检测 Jurkat 和 Raji 细胞中 p27^(kip1)与 Jab1的结合情况。结果血清饥饿导致 Jurkat 和 Raji 细胞生长周期停滞,p27^(kip1)蛋白总量增加,Jab1蛋白总量减少。血清释放后两者的蛋白水平呈现相反的表达变化,而 p27^(kip1)mRNA 水平无明显改变。LMB 可以抑制由血清释放引起的细胞增殖,在此过程中 p27^(kip1)表达上调,Jab1表达下调。转染 Jab1真核表达质粒的 Jar-kat 细胞 p27^(kip1)定位有明显的改变。免疫沉淀结果显示在 Jurkat 和 Raji 细胞中 p27^(kip1)与 Jab1相互结合。结论 Jab1可能通过与 p27^(kip1)结合来介导 p27^(kip1)的核内外分布并影响其表达,进而影响 p27^(kip1)的功能状态,从而参与调控淋巴瘤细胞的生长。Objective To investigate the expression and relationship of p27^kip1 and its nuclear export factor Jab1 during proliferation process of lymphoma cell. Methods Jurkat and Raji cells were treated with serum starvation and then serum release. The protein and mRNA expression of p27^kip1 ,Jab1 in the cells were detected by Western blot and RT-PCR respectively. LMB were used for stimulating Jurkat cells during their proliferation process, and then the expression changes of p27^kip1 and Jab1 were detected. An eukaryotic expression plasmid（pcDNA3.1-myc） containing Jab1 was contructed. Jurkat cell were transfected in vitro with or without pcDNA3.1-myc-Jab1. Double immunolabelling was used to identify the localization of p27^kip1. Immunoprecipitation was used to detect the combination of p27^kip1 and Jab1. Results The growth of Jurkat and Raji cells were blocked by serum starvation. The total protein amount of p27^kip1 increased while that of Jab1 decreased. The reverse changes were happened after serum release, but the mRNA expression of p27^kip1 has no significant change. LMB could inhibit the cell proliferation caused by serum release. The expression of p27^kip1 was up-regulated and Jab1 down-regulated when Jurkat cells were treated with LMB. After pcDNA3.1- myc-Jab1 infected Jurkat cells for 48 h, the distribution of p27^kip1 was translocated from nucleus into cytoplasma. p27^kip1 and Jab1 could form compound in Jurkat and Raji cells detected by Immunoprecipitation. Conclusion Jab1 may influence the location and expression of p27^kip1 through integrating with p27^kip1 , and then participates in regulating the growth of NHL cell through interfering with the function of p27^kip1.

关 键 词：细胞系 Jurkat 细胞系 Raji p27p27^kip1蛋白 激活蛋白1辅因子 

分 类 号：R733.1[医药卫生—肿瘤]