CD74基因表达对急性髓系白血病细胞增殖与侵袭的抑制作用

王筝.CD74基因表达对急性髓系白血病细胞增殖与侵袭的抑制作用[J].内科急危重症杂志,2023,29(2):146-150

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DOI：10.11768/nkjwzzzz20230212

中文关键词: 急性髓系白血病白细胞分化抗原74 细胞增殖细胞侵袭巨噬细胞移动抑制因子/细胞外信号调节激酶信号通路

英文关键词:

基金项目:北京市科技计划项目（No: Z171100001017103）

作者	单位	E-mail
王筝	航天中心医院血液科	wangjingbo29@sohu.com

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中文摘要:

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LipofectamineTM2000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、72h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、72h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。

英文摘要:

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 lipofectamineTM2000 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 OD450 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.