江苏大学学报（医学版）2025，Vol.35Issue(6)：474-479,495,7.DOI:10.13312/j.issn.1671-7783.y240182
rL-RVG通过调节小鼠巨噬细胞Raw264.7极化抑制小鼠Lewis肺癌细胞活力
rL-RVG inhibits the viability of Lewis lung cancer cells by regulating the polarization of mouse macrophage Raw264.7
摘要
Abstract
Objective:To explore the effects of recombinant avirulent Newcastle disease virus La Sota strain expressing rabies virus glycoprotein(rL-RVG)on the polarization of murine macrophage RAW264.7 cells and to analyze the effect of polarized RAW264.7 cells on the viability of Lewis lung cancer cells.Methods:Murine macrophage RAW264.7 cells were divided into four groups:blank control group and rL-RVG groups at different multiplicities of infection(0.5,1,5 MOI);and the viability of RAW264.7 cells was assessed by using the CCK-8 assay.RAW264.7 cells were divided into three groups:RAW264.7-M0 type(complete media),RAW264.7-M1 type(induced by 1 μg/mL lipopolysaccharide+50 ng/mL IFN-γ),and RAW264.7-M2 type(induced by 20 ng/mL IL-4);and the morphological characteristics of these phenotypes were observed under phase-contrast microscopy.The protein expression levels of CD86 and Arg-1 were measured by immunofluorescence in four groups:Raw264.7-M1,Raw264.7-M2,rL-RVG+Raw264.7-M0,and rL-RVG+Raw264.7-M2.The mRNA expression levels of cytokines IL-4,IL-6,IL-10 and tumor necrosis factor-α(TNF-α)in the blank control group and rL-RVG groups were also assessed by qRT-PCR.Lewis lung cancer cells were co-cultured with conditioned media from the above groups and divided into four groups:blank control group and rL-RVG groups at different MOIs(0.5,1,5 MOI).The viability of Lewis lung cancer cells was evaluated by using the CCK-8 assay.Results:Compared with the blank control group,the cell proliferation rates in the 1,5 MOI rL-RVG groups were significantly increased(both P＜0.01).The morphological characteristics of Raw264.7-M0,M1 and M2 macrophages were round or oval,flat or irregular,and fibrous,elongated or spindle-shaped,respectively.Compared with RAW264.7-M0 type macrophages,the relative expression level of CD86 mRNA in RAW264.7-M1 type was significantly increased(P＜0.01),and the relative expression level of Arg-1 mRNA in RAW264.7-M2 type was greatly increased(P＜0.01).Compared with Raw264.7-M1 type,the relative fluorescence expression level of CD86 in rL-RVG+Raw264.7-M0 type was significantly increased(P＜0.05);compared with Raw264.7-M2 type,the relative fluorescence expression level of CD86 in rL-RVG+Raw264.7-M2 type was significantly increased(P＜0.05),and the relative fluorescence expression level of Arg-1 was significantly decreased(P＜0.05).rL-RVG treatment led to an increase in the mRNA expressions of TNF-α and IL-6,while the mRNA expressions of IL-4 and IL-10 decreased.Compared with the blank control group,the viability of Lewis lung cancer cells in the 0.5,1,and 5 MOI rL-RVG groups were significantly decreased(all P＜0.01).Conclusion:rL-RVG could induce the polarization of RAW264.7 macrophages towards the M1 phenotype and subsequently inhibit the viability of Lewis lung cancer cells at MOIs ranging from 0.5 to 5.关键词
稳定表达狂犬病病毒糖蛋白的重组新城疫病毒/肺腺癌/巨噬细胞/表型/极化Key words
recombinant avirulent Newcastle disease virus La Sota strain expressing the rabies virus glycoprotein/lung adenocarcinoma/macrophages/phenotypes/polarization分类
医药卫生引用本文复制引用
孙家卿,严玉兰..rL-RVG通过调节小鼠巨噬细胞Raw264.7极化抑制小鼠Lewis肺癌细胞活力[J].江苏大学学报（医学版）,2025,35(6):474-479,495,7.基金项目
深圳医学研究专项资金(C2401017) （C2401017）
