MEF2A激活p38 MAPK信号通路促进骨髓增殖性肿瘤细胞增殖及血管形成  

MEF2A promotes the proliferation and angiogenesis of myeloproliferative neoplasm by activating p38 MAPK signaling pathway

作　　者：唐荣芳[1] 张颖杰 徐淑娟 孔令珍 聂宇薇[1] TANG Rong-fang;ZHANG Ying-jie;XU Shu-juan;KONG Ling-zhen;NIE Yu-wei(Department of Hematology,Affiliated Hospital of Guilin Medical University,Guilin 541001,China)

机构地区：[1]桂林医学院附属医院血液科,桂林541001

出　　处：《现代免疫学》2024年第3期243-250,共8页Current Immunology

基　　金：广西区卫生和计划生育委员会自筹经费科研课题(Z201708)。

摘　　要：为探讨肌细胞增强因子2A(myocyte enhancer factor 2A,MEF2A)基因在骨髓增殖性肿瘤(myeloproliferative neoplasm,MPN)中的表达及其与细胞增殖、血管生成的关系,收集2020年1月至2021年12月桂林医学院附属医院30例同期体检健康志愿者和60例MPN患者的PBMC;另选MPN细胞系HEL92.1.7(简称HEL)、UKE-1和SET-2进行细胞培养。采用qRT-PCR法、Western blotting检测各细胞中MEF2A的mRNA和蛋白表达水平。选取HEL和UKE-1细胞,分别转染MEF2A敲减/过表达质粒。采用CCK-8法及结晶紫染色测定细胞活力及单克隆形成能力;采用AnnexinⅤ-FITC/PI双染流式细胞术测定细胞凋亡率;导管形成实验测定细胞体外促血管形成能力;Western blotting测定肿瘤细胞凋亡、血管形成以及MAPK信号通路相关蛋白的表达。结果显示,MPN患者PBMC中MEF2A的mRNA和蛋白表达量均显著高于健康志愿者(均P<0.05);MEF2A的mRNA和蛋白在各MPN细胞系中的表达量均显著高于其在健康志愿者PBMC中的表达量(均P<0.05)。与Control组比较,MEF2A过表达可提高HEL和UKE-1细胞的增殖率、单克隆形成数目、血管成管数量及ANGPT2、FGF1、PDGFA、VEGF、Bcl-2、p-P38和p-ERK蛋白表达量(均P<0.05),降低HEL和UKE-1细胞凋亡率及BAX、剪切caspase 3(cleaved caspase 3,cl-caspase-3)蛋白表达量(均P<0.05)。MEF2A敲减可抑制HEL和UKE-1细胞的增殖率、单克隆形成数目、血管成管数量及ANGPT2、FGF1、PDGFA,VEGF、Bcl-2、p-P38和p-ERK的蛋白表达量(均P<0.05),提高HEL和UKE-1细胞凋亡率及BAX、cl-caspase-3蛋白表达量(均P<0.05)。由此,MEF2A在MPN中高表达,敲减MEF2A可抑制MPN细胞增殖及血管形成,促进细胞凋亡,其机制可能与调节MAPK信号通路有关。The aim of this paper was to investigate the expression of myocyte enhancer factor 2A(MEF2A)gene in myeloproliferative neoplasm(MPN)and its relation with tumor cell proliferation and angiogenesis.From January 2020 to December 2021,PBMC specimens were collected from 30 healthy volunteers and 60 MPN patients in the Affiliated Hospital of Guilin Medical University.Additionally,MPN tumor cell lines HEL92.1.7(abbreviated HEL),UKE-1,and SET-2 were cultured and used for cellular study.qRT-PCR and western blotting were used to detect the expression levels of MEF2A mRNA and protein.HEL and UKE-1 cells were transfected with MEF2A knock-down or over-expressing plasmid,respectively.Cell viability and monoclonal formation capacity were evaluated by CCK-8 assay and crystal violet staining.Cell apoptosis rate was measured by FACS with AnnexinⅤ-FITC/PI double staining.The in vitro angiogenic ability was determined by catheter formation assay.The expressions of proteins related to apoptosis,angiogenesis and p38 MAPK signaling pathway were quantified by western blotting.The results showed that the mRNA and protein expressions of MEF2A in PBMCs of MPN patients were significantly higher than those of healthy volunteers(both P<0.05).The expression levels of MEF2A mRNA and protein in MPN cell lines were also significantly higher than those of PBMCs from healthy controls(all with P<0.05).In HEL and UKE-1 cell lines,MEF2A over-expression caused increased cell proliferation,clone formation,angiogenesis,and the protein expressions of ANGPT2,FGF1,PDGFA,VEGF,Bcl-2,p-P38,and p-ERK(all with P<0.05).In contrast,it decreased the cell apoptosis rate and the protein expressions of BAX and cleaved caspase 3(cl-caspase-3)(all with P<0.05).On the other hand,MEF2A knock-down resulted in the complete opposite effects(all with P<0.05).In conclusion,MEF2A is broadly distributed in MPN cells at a relatively high level,and knock-down of MEF2A can inhibit the proliferation and angiogenesis while promoting the apoptosis of MPN tumor cells.MEF2A may functi

关 键 词：骨髓增殖性肿瘤 肌细胞增强因子2A 丝裂原激活的蛋白激酶信号通路 细胞增殖 血管形成 

分 类 号：R733.3[医药卫生—肿瘤]