人卵巢癌荷瘤裸鼠肿瘤微环境局部F4/80阳性巨噬细胞表达Foxp3

Foxp3 was expressed in F4/80+macrophages in tumor microenvironment of nude mice bearing human ovarian cancer

目的:探讨人卵巢癌裸鼠腹腔荷瘤模型体内F4/80阳性巨噬细胞是否表达Foxp3,初步探索卵巢癌改造免疫微环境促进自身发展的新机制。方法:应用人卵巢癌细胞系SKOV3构建人卵巢癌裸鼠腹腔荷瘤模型为实验组,以未处理裸鼠为对照组,流式细胞术检测两组外周血和腹水中F4/80阳性巨噬细胞Foxp3的表达情况。应用流式细胞分选两组腹水中F4/80阳性巨噬细胞,分别应用Western blot和PCR检测该细胞中Foxp3在蛋白和mRNA水平的表达情况。免疫荧光双染法鉴定腹腔内卵巢癌组织局部浸润F4/80阳性巨噬细胞Foxp3的表达情况。结果:流式细胞术检测两组外周血中F4/80阳性细胞均不表达Foxp3,差异无统计学意义(P>0.05)。流式细胞术和Western blot显示实验组腹水F4/80阳性巨噬细胞在蛋白水平明显表达Foxp3,实时荧光定量PCR显示实验组在mRNA水平明显表达Foxp3,与对照组比较差异均有统计学意义(P<0.05)。免疫荧光双染法显示卵巢癌组织可见F4/80和Foxp3双阳性细胞。结论:卵巢癌肿瘤微环境中F4/80阳性巨噬细胞表达Foxp3,为寻找卵巢癌免疫疗法新靶点奠定基础。

Objective:To investigate the expression of Foxp3 in F4/80+macrophages of nude mice bearing human ovarian cancer,and to explore the new mechanism of remoulding the immune microenvironment and promoting its own development.Methods:Establishing the human ovarian cancer xenograft model in nude mice by human ovarian cancer cell line SKOV3 as experimental group,the untreated nude mice was control group.Flow cytometry was used to detect the expression of Foxp3 in F4/80+macrophages in peripheral blood and ascites.Flow cytometry was used to isolate F4/80 positive macrophages from ascites,then Western blot and PCR were used to detect the expression of Foxp3 at protein and mRNA levels.The expression of Foxp3 in F4/80+macrophages infiltrated into ovarian cancer tissue in abdominal cavity was identified by double immunofluorescence staining.Results:Foxp3 was not detected in F4/80+cells in peripheral blood of the two groups by flow cytometry(P>0.05).The results of Flow cytometry and Western blot showed that F4/80+macrophages in ascites of the experimental group expressed Foxp3 at protein level and real-time fluorescence quantitative PCR showed that Foxp3 was expressed at mRNA level in the experimental group,which was significantly different from the control group(P<0.05).Immunofluorescence staining showed that F4/80 and Foxp3 double positive cells could be seen in ovarian cancer tissues.Conclusion:Foxp3 has been expressed in F4/80+macrophages in ovarian cancer microenvironment,which laid a foundation for finding new targets of immunotherapy for ovarian cancer.