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目的探讨p27kip1及相关分子Jab1、CRM1在淋巴瘤细胞U937增殖过程中的表达及相互关系。方法采用血清饥饿合并释放处理淋巴瘤细胞株U937,通过细胞计数法检测该处理对U937细胞生长情况的影响;采用Western blot、免疫荧光双标法和激光共聚焦技术检测不同生长状态的U937细胞中p27kip1、Jab1的表达及亚细胞定位情况。结果血清饥饿造成U937细胞生长阻滞, p27kip1相关分子Jab1和CRM1表达下降,p27kip1蛋白总量增加,第10位丝氨酸磷酸化的p27kip1表达下降。与此同时,p27kip1也由胞浆高分布转变为胞核高分布。血清释放后上述分子呈现相反的变化,并且p27kip1的分布又主要集中在胞浆中。结论在刺激淋巴瘤细胞U937增殖过程中,Jab1和CRM1可能通过影响p27kip1的亚细胞定位与表达水平,参与调控淋巴瘤细胞的生长状态。
Objective To investigate the expression and correlation of p27kip1 and related molecules Jab1 and CRM1 in the proliferation of lymphoma U937 cells. Methods U937 cells were treated with serum-starvation combined with release. U937 cells were treated with U937 cells by cytometry. Western blot, double immunofluorescence and confocal laser scanning microscopy were used to detect the expression of U937 cells in different growth states p27kip1, Jab1 expression and subcellular localization. Results Serum starvation induced the growth arrest of U937 cells, decreased the expression of p27kip1-related molecules Jab1 and CRM1, increased the total amount of p27kip1 protein and decreased the phosphorylation of p27kip1 at serine 10. At the same time, p27kip1 is also changed from highly cytoplasm to highly nuclear. The above molecules showed the opposite change after serum release, and the distribution of p27kip1 mainly concentrated in the cytoplasm. Conclusions Jab1 and CRM1 may be involved in the regulation of the growth of lymphoma cells by affecting the subcellular localization and expression of p27kip1 in the process of stimulating the proliferation of U937 lymphoma cells.