摘要
目的探究DIS3对人骨髓瘤细胞周期及肿瘤相关蛋白表达的影响。方法选取人骨髓瘤细胞株NCI-H929、RPMI 8226和U266为研究对象,分别设计并构建DIS3基因过表达载体和DIS3-siRNA。PI单染检测细胞周期变化,qRT-PCR及Western blot检测周期相关蛋白Cyclin B1、P21、CDK2以及肿瘤相关蛋白MYC、RAS、TP53及BRAF的表达。同时,采用Western blot检测ERK1/2信号通路激活水平。结果与空载体组比较,DIS3过表达组细胞存在明显的G0/G1期阻滞,周期相关蛋白Cyclin B1、CDK2以及肿瘤相关蛋白MYC、RAS、TP53、BRAF和p-ERK1/2的表达水平下降(P<0.05,P<0.01),P21表达水平升高(P<0.01),siRNA组各指标变化趋势与DIS3过表达组相反。结论DIS3过表达能使人骨髓瘤细胞发生明显的G0/G1期阻滞,降低肿瘤相关蛋白的表达,这可能与抑制ERK1/2信号通路的激活密切相关。
Objective To observe the effects of DIS3 overexpression or interference on expression of cell cycle and tumor-associated proteins in three types of human myeloma cells.Methods Human myeloma cell lines NCI-H929,RPMI 8226,and U266 were selected as study objects.A DIS3 gene overexpression vector and DIS3-siRNA were designed and constructed.Cell cycle was evaluated by propidium iodide single staining.qRT-PCR and western blotting were employed to detect expression of cyclin B1,P21,CDK2,and tumor-associated proteins MYC,RAS,TP53,and BRAF at mRNA and protein levels,respectively.In addition,activation of the ERK1/2 signaling pathway was detected by Western blot.Results Compared with the empty vector group,cells in the DIS3 overexpression group exhibited obvious G0/G1 phase arrest and decreased expression levels of cycle-related proteins cyclin B1 and CDK2,as well as tumor-related proteins MYC,RAS,TP53,BRAF,and p-ERK1/2(P<0.05,P<0.01).The expression of P21 was increased(P<0.01)in DIS3 overexpression group.In the siRNA group,trends for each index of mRNA expression or protein expression were opposite to those observed in the DIS3 overexpression group.Conclusions DIS3 overexpression can significantly block G0/G1 phase in human myeloma cells and reduce expression of tumor-associated proteins,which may be closely related to the inhibition of ERK1/2 signaling pathway activation.
作者
邹亮
程辉
张婷
周英
关军
程平
王兰兰
ZOU Liang;CHENG Hui;ZHANG Ting;ZHOU Ying;GUAN Jun;CHENG Ping;WANG Lanlan(Department of Hematology,Wuhan First Hospital,Wuhan 430022,China)
出处
《中国比较医学杂志》
CAS
北大核心
2020年第11期40-46,共7页
Chinese Journal of Comparative Medicine
基金
武汉市卫计委基金(WX17C05)。
