卵巢癌细胞系SKOV3培养上清对巨噬细胞共刺激分子的影响

Effects of culture supernatant of ovarian cancer cell line-SKOV3 on costimulatory molecules of macrophages

目的研究卵巢癌细胞系SKOV3培养上清处理巨噬细胞后,巨噬细胞亚型的改变,以及CD80、CD86、CD40在不同巨噬细胞亚型上表达的差异。方法 Ficoll密度梯度法分离健康成人外周血单个核细胞,GM-CSF诱导为巨噬细胞(M0)后,用对数生长期的SKOV3细胞培养上清处理14 d后,采用流式细胞术检测巨噬细胞亚型的改变及不同亚型细胞CD80、CD86和CD40的表达情况。结果流式细胞术检测显示,用SKOV3细胞培养上清培养巨噬细胞14 d后,巨噬细胞主要向CD163-M2型巨噬细胞分化[M1/M2=(12.37±1.76)%/(22.23±2.21)%]。CD163-M2型巨噬细胞CD40的表达较CD64-M1型显著降低,2组比较差异均有统计学意义(P<0.05);而CD80、CD86的表达无明显改变,2组比较差异无统计学意义(P>0.05)。结论 SKOV3细胞培养上清刺激后的巨噬细胞向不同巨噬细胞亚型分化;M2型巨噬细胞较M1型巨噬细胞共刺激分子表达下调,提示这些共刺激分子在巨噬细胞的活化及免疫应答过程中很重要,可能与肿瘤细胞的免疫逃逸有关。

Objective To investigate the changes of subtypes of macrophage treated by culture supernatant of ovarian cancer cell line-SK0V3 in vitro,and to explore the differences of expressions of CD80, CD86, CD40 in different subtypes of macrophage. Methods The peripheral blood mononuclear cells were separated from healthy adults by Ficoll density gradient method,which were induced into macrophages by GM-CSF. Then the macrophages were cultured in supernatant of SK0V3 ovarian cancer cells at exponential growth phase for 14 days. The changes of subtypes of macrophage and the expression levels of CD80, CD86, CD40 in different subtypes of macrophages were detected by flow cytometry. Results The examination results by flow cytometry showed that after the macrophages were treated in supernatant of SK0V3 ovarian cancer cells for 14 days, the macrophages were mainly differentiated into CD163-M2 subtype macrophages [ M1/M2 = （ 12. 37 ± 1. 76） %/（22. 23 土 2.21）%]. The expression levels of CD40 in CD163-M2 subtype macrophages were significantly decreased,as compared with those in CD64-M1 subtype macrophage （ P 〈0.05 ） . However there were no significant differences in the expression levels of CD80 and CD86 between two groups （ P 〉 0.05 ） . Conclusion After the macrophages are treated in culture supernatant of SK0V3 ovarian cancer cells, which are differentiated into different subtypes. Moreover the expression levels of costimulatory molecules are down - regulated in M2 subtype macrophages, as compared with those in Ml subtype macrophages,which suggests that these costimulatory molecules play an important role in activation of macrophages and immune response, which may be correlated to immune escape of tumor cells.