RNA干扰PNAS-2后U937细胞周期及相关蛋白表达的变化

Changes of cell cycle and related proteins after suppression of PNAS-2 gene expression by RNA interference in cell line U937

目的研究RNA干扰抑制细胞凋亡相关蛋白(PNAS-2)基因表达后,急性单核细胞白血病细胞株U937细胞周期及其相关蛋白表达的变化。方法将已构建的靶向抑制PNAS-2的干扰组质粒和对照组质粒分别转染U937细胞。流式细胞仪分析两组细胞周期的变化;蛋白质芯片技术比较两组细胞周期相关蛋白表达水平的差异。结果流式细胞仪检测结果显示,干扰组细胞出现G2/M期阻滞,与对照组比较差异有统计学意义;蛋白质芯片结果分析发现,有29条细胞周期相关蛋白表达变化,其中上调25条(cyclinA、cyclinE及相关调节因子p53蛋白等),下调4条。CyclinB表达水平无显著改变。结论RNA干扰抑制PNAS-2表达后,U937细胞周期出现G2/M期阻滞,可能与细胞周期素A、p53蛋白表达上调,而细胞周期素B表达不变有关。

Objective To study the changes of cell cycle and related proteins after suppression of apoptosis-related protein- 2 （PNAS-2） gene expression by RNA interference in acute monocytic leukemia cell line U937. Methods Cell lines U937 were transfecteds with plamids in control group and constructed plasmids for PNAS-2 suppression in interference group, respectively. The changes of cell cycle were analysed by flow cytometry, and the expressions of cell cycle-related proteins were compared between these two groups by protein microchip technique. Results It was indicated by flow cytometry that cell arrest appeared in G2/M phase in interference group, and there was significant difference between control group and interference group. The changes of expressions of 29 cell cycle-related proteins were detected by protein microchip, and it was revealed that the expressions of 25 proteins such as cyclin A, cyclin E and p53 protein were up-regulated, while the other 4 were down-regulated. There was no significant change in the expression of cyclin B. Conclusion Suppression of PNAS-2 expression by RNA interference may lead to G2/M phase arrest in cell line U937, which may be related to the upregulation of expressions of cyclin A and p53 protein and unchanged expression of cyclin B.