摘要
目的探讨RNA甲基化酶NOP2/Sun结构域家族成员2(NSUN2)调控巨噬细胞M2极化的作用机制。方法用小干扰RNA(siRNA)干扰技术敲低鼠源巨噬细胞RAW264.7中NSUN2后,分别用RT-PCR和流式细胞术检测细胞向M1和M2极化情况,CCK-8试剂检测膀胱癌细胞生长情况,Western blot实验检测细胞因子信号抑制因子(SOCS)3/JAK2/STAT3信号通路蛋白表达水平的变化情况。同时敲低SOCS3,Western blot实验检测JAK2/STAT3通路蛋白表达水平的变化情况。结果敲低NSUN2后,鼠源巨噬细胞RAW264.7内M1极化相关基因肿瘤坏死因子-α(TNF-α)、IL-6、一氧化氮合酶2(NOS2)mRNA表达水平明显增加,细胞向M1极化的比例增高,且其促进膀胱癌MB49细胞生长的能力降低。同时,敲低NSUN2后,IL-4诱导鼠源巨噬细胞RAW264.7向M2极化的能力下降,SOCS3蛋白表达水平升高,JAK2、STAT3蛋白水平及磷酸化水平下降;若敲低SOCS3表达,鼠源巨噬细胞RAW264.7内JAK2和STAT3蛋白磷酸化水平升高。结论NSUN2调控SOCS3/JAK2/STAT3信号通路促进巨噬细胞向M2极化。
Objective To investigate the mechanism by which the RNA methyltransferase NOP2/Sun domain family member 2(NSUN2)regulates M2 polarization of macrophages.Methods Following knockdown of NSUN2 in murine macrophage RAW264.7 cells using siRNA interference technology,M1 and M2polarization was assessed by RT-PCR and flow cytometry,respectively.The growth of bladder cancer cells was measured with the CCK-8 assay.Changes in the expression levels of proteins in the cytokine signaling suppressor(SOCS)3/JAK2/STAT3 signaling pathway were detected by Western blot.Additionally,SOCS3 was knocked down,and alterations in JAK2/STAT3 pathway protein expression were examined by Western blot.Results After knockdown of NSUN2,the expression levels of M1 polarization-related genes,including tumor necrosis factor-α(TNF-α),IL-6,and NOS2 mRNA,in mouse-derived macrophages RAW264.7 were significantly increased,and the proportion of cells undergoing M1 polarization was elevated.Additionally,the ability of mouse-derived macrophages RAW264.7 to promote the growth of bladder cancer MB49 cells was reduced.Simultaneously,after knockdown of NSUN2,the ability of IL-4 to induce mouse-derived macrophages RAW264.7 to undergo M2 polarization decreased,the expression level of SOCS3 protein increased,and the levels and phosphorylation of JAK2 and STAT3 proteins decreased;if SOCS3 expression was knocked down,the phosphorylation levels of JAK2 and STAT3 proteins in mouse-derived macrophages RAW264.7 increased.Conclusion NSUN2 promotes M2 polarization of macrophages by regulating the SOCS3/JAK2/STAT3 signaling pathway.
作者
韩娜
唐懿
邓鑫霈
袁刚军
HAN Na;TANG Yi;DENG Xinpei;YUAN Gangjun(Health Examination and Oncology Screening Center,Chongqing University Cancer Hospital,Chongqing 400030,China;Department of Urology,Sun Yat-sen University Cancer Center,Guangzhou,Guangdong 510060,China;Department of Urology,Chongqing University Cancer Hospital,Chongqing 400030,China)
出处
《重庆医学》
2026年第3期515-520,共6页
Chongqing Medical Journal
基金
重庆市科研机构绩效激励引导专项(cstc2022jxjl120029)。
