p27kip1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

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^kip1mRNA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Jurkat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。

Expression and relationship of p27kip1 and it's nuclear export factor Jab1 in Lymphoma cell Jurkat

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).