摘要
目的:探讨白细胞分化抗原74(CD74)基因表达对急性髓系白血病(AML)细胞增殖、侵袭及巨噬细胞移动抑制因子(MIF)/细胞外信号调节激酶(ERK)信号通路的影响。方法:收集83例AML患者的骨髓样本(AML组)、25例正常供者的骨髓样本(对照组)及人AML细胞系HL60、MOLM-13、THP-1、U-937、人原代正常骨髓单个核细胞(BMMC)为研究对象,Western blot检测AML骨髓样本及各细胞系中CD74蛋白表达;利用Lipofectamine ^(TM) 2000转染试剂盒对MOLM-13细胞进行转染,并分为空白组(细胞不进行转染)、si-NC组(将si-NC转染于细胞)、si-CD74组(将si-CD74转染于细胞);Western blot检测各组细胞中CD74、基质金属蛋白酶9(MMP-9)及MIF/ERK信号通路相关蛋白MIF、磷酸化细胞外信号调控蛋白激酶(p-ERK)、ERK表达水平;CCK-8法检测各组MOLM-13细胞在0、24、48、72 h的增殖情况;Transwell实验检测各组MOLM-13细胞侵袭情况。结果:与对照组比较,AML组骨髓细胞中CD74蛋白表达水平显著升高(P<0.05),CD74蛋白在HL60、MOLM-13、THP-1、U-937细胞系中均高表达,且在MOLM-13细胞系中表达量最高。与空白组和si-NC组比较,si-CD74组MOLM-13细胞的OD450值(在24、48、72 h时)、细胞侵袭数目、CD74、MMP-9、MIF和p-ERK蛋白表达显著降低(P均<0.05),但空白组与si-NC组比较,差异无统计学意义(P均>0.05)。结论:干扰AML细胞中CD74表达,可抑制细胞增殖与侵袭,其作用机制可能与抑制MIF/ERK信号通路有关。
Objective:To investigate the effects of leukocyte differentiation antigen 74(CD74)gene expression on the proliferation,invasion and macrophage migration inhibitory factor(MIF)/extracellular signal-regulated kinase(ERK)signaling pathway in acute myeloid leukemia(AML)cells.Methods:Bone marrow samples from 83 AML patients(AML group),bone marrow samples from 25 normal donors(control group)and human AML cell lines HL60,MOLM-13,THP-1,U-937,and bone marrow mononnuclear cell(BMMC)were collected as the research objects.Western blotting was used to detect the CD74 protein expression in AML bone marrow samples and cell lines.MOLM-13 cells were transfected with lipofectamine^(TM)2000 transfection kit and divided into blank group(without transfecting),si-NC group(transfecting with si-NC),si-CD74 group(transfection with si-CD74).Western blotting was used to detect the expression levels of CD74,matrix metalloproteinase 9(MMP-9)and MIF/ERK signaling pathway related proteins MIF,p-ERK and ERK in each group of cells,CCK-8 method was used to detect the proliferation of MOLM-13 cells in each group at 0,24,48,72 h,and Transwell experiment was used to detect MOLM-13 cell invasion in each group.Results:Compared with the control group,the expression level of CD74 protein in the bone marrow of the AML group was significantly increased(P<0.05).CD74 protein was highly expressed in HL60,MOLM-13,THP-1,U-937 cell lines,and was highest in MOLM-13 cell line.Compared with the blank group and the si-NC group,the OD 450 value(at 24,48,72h),cell invasion number,expression of CD74,MMP-9,MIF and p-ERK proteins in MOLM-13 cells in the si-CD74 group were significantly reduced(P<0.05).There was no significant difference between the blank group and the si-NC group(P>0.05).Conclusion:Interfering with the expression of CD74 in AML cells can inhibit cell proliferation and invasion.This mechanism may be related to the inhibition of MIF/ERK signaling pathway.
作者
王筝
王静波
李静静
万春超
张岚
WANG Zheng;WANG Jing-bo;LI Jing-jing;WAN Chun-chao;ZHANG Lan(Department of Hematology,Aerospace Center Hospital,Beijing 100049,China)
出处
《内科急危重症杂志》
2023年第2期146-150,共5页
Journal of Critical Care In Internal Medicine
基金
北京市科技计划项目(No:Z171100001017103)。
