广西医科大学学报2023，Vol.40Issue(12)：1933-1940,8.DOI:10.16190/j.cnki.45-1211/r.2023.12.001
LCMT1过表达对M2型巨噬细胞极化的影响
Effect of leucine carboxyl methyltransferase 1 overexpression on polarization of M2 macro-phages
摘要
Abstract
Objective:To establish the THP-1 cell lines with leucine carboxyl methyltransferase 1(LCMT1)overexpression by lentiviral vector and study the regulating effect of LCMT1 on THP-1 cells and M2 polarization of THP-1-derived macrophages.Methods:Lentiviral technology was used for the construction of LCMT1 over-expression vectors.Blank group,OE-control group,and OE-LCMT1 group were set up.The green fluorescence of each group of cells was observed by fluorescence microscope.The proliferation levels of each group of cells were detected by the cell counting kit-8(CCK-8)assay.Real-time fluorescence quantitative PCR(RT-qPCR)and western blotting were carried out to evaluate LCMT1 mRNA and protein expression level,and the expression lev-el of PP2Ac methylation-associated protein.THP-1 stable transformation cells were treated by 100 nmol/L phor-bol myristate acetate(PMA)for 48 hours and induced to polarize into M0 macrophages,and M0 macrophages were induced to polarize into M2 macrophages by 20 ng/mL interferon-4(IL-4)for 48 hours.The mRNA expression of mannose receptor C type 1(MRC-1),C-C motif chemokine ligand 22(CCL22)and dendritic cell-specific ICAM-3 grabbing non-integrin(CD209)in the polarization markers of M2 macrophages were detected by RT-qPCR.Results:Compared with the blank group,the expression of green fluorescence was expressed in the OE-control group and the OE-LCMT1 group,the proliferation curve was decreased in the OE-control group(P＜0.001),and the expression levels of LCMT1 and PP2Ac methylation-associated protein differences were not statistically significant in the OE-control group(P＞0.05).Compared with the OE-control group,the proliferation curve of the OE-LCMT1 group was decreased(P＜0.05),the expression of LCMT1 and PP2Ac methylated proteins was increased(P＜0.05),the expression of PPME-1 and PP2Ac demethylated proteins was decreased(P＜0.01)and the expression of total PP2Ac protein differences was not statistically significant in the OE-LCMT1 group(P＞0.05).Compared with the blank M0 group,the mRNA expression levels of MRC-1,CD209,and CCL22of M2 polarization markers were increased in the blank M2 group(P＜0.05).Compared with the OE-control M2 group,the mRNA expression levels of MRC-1,CD209,and CCL22of M2 polarization markers were reduced in the OE-LCMT1 M2 group(P＜0.05).Conclu-sion:The THP-1 cell lines with LCMT1 overexpression are successfully constructed.LCMT1 overexpression can inhibit the proliferation of THP-1 cells,maintain the total PP2Ac protein levels,enhance the PP2Ac methyla-tion with concomitant reduction in PPME-1 and PP2Ac demethylation,down-regulate the polarization markers of M2 macrophages,and inhibit the polarization of M2 macrophages.关键词
亮氨酸羟基甲基转移酶1/人髓系白血病单核细胞/慢病毒载体/M2型巨噬细胞/巨噬细胞极化Key words
leucine carboxyl methyltransferase 1/human myeloid leukemia mononuclear cells/lentiviral vector/M2 macrophages/macrophage polarization分类
医药卫生引用本文复制引用
李菡,唐深,蓝利,王新航,刘彧冰,王名鸿,袁江浪,孔星星,陆彩玲,李习艺..LCMT1过表达对M2型巨噬细胞极化的影响[J].广西医科大学学报,2023,40(12):1933-1940,8.基金项目
国家自然科学基金项目(No.82260320 （）
No.82160612) （）
广西自然科学基金项目(No.2021GXNSFAA196019) （No.2021GXNSFAA196019）
