卵巢癌SKOV3细胞系培养上清液处理巨噬细胞不同时间后对巨噬细胞分化的影响  

Effects of macrophage treated by culture supernatant with ovarian cancer SKOV3 cell line at different time points on the differentiation of macrophages

作　　者：宋玉霞[1] 赵涛[1] 张玮璨 郭清[1] 王宏卫[1] 孟桐羽[1] 

机构地区：[1]河北省石家庄市第一医院妇产科,050011 [2]河北医科大学

出　　处：《河北医药》2016年第10期1498-1501,1504,共5页Hebei Medical Journal

摘　　要：目的探讨卵巢癌SKOV3细胞系培养上清液处理巨噬细胞不同时间后,巨噬细胞分化亚型的改变。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细胞培养上清液处理为实验组,用1640培养基处理为对照组,分别于培养第7天和第14天光学显微镜下观察巨噬细胞形态变化;流式细胞术检测实验组和对照组在不同时间后巨噬细胞表面细胞因子(CD64、CD163)表达情况。结果培养第7天时,实验组巨噬细胞形态改变明显,细胞突起增加,细胞变细长,对照组细胞形态无明显变化,实验组CD64-M1型巨噬细胞所占比例高于对照组,CD163-M2型巨噬细胞所占比例低于对照组(P<0.05);培养第14天时,实验组巨噬细胞形态逐渐恢复,对照组细胞形态仍无明显变化,实验组CD64-M1型巨噬细胞所占比例低于对照组,CD163-M2型巨噬细胞所占比例高于对照组(P<0.05)。实验组CD64-M1型巨噬细胞在第7天时M1/M0水平高于第14天,CD163-M2型巨噬细胞在第7天时M2/M0低于在第14天,差异均有统计学意义(P<0.05)。结论 SKOV3细胞培养上清液可改变巨噬细胞形态,激活巨噬细胞免疫反应,随着时间的推移,细胞形态恢复,发生免疫抑制;同时,也可促进巨噬细胞分化,随着时间的推移,亚型也有不同,主要由CD64-M1型巨噬细胞向CD163-M2型巨噬细胞转变。Objective To investigate the changes of differentiation subtypes of macrophages treated by culture supernatant with ovarian cancer SKOV3 cell line at different time points. Methods The peripheral blood mononuclear cells were separated from healthy adults by Ficoll density gradient method,which were induced into macrophages（ M0） by GM-CSF. The culture supernatant in which the macrophages were cultured in medium with SKOV3 ovarian cancer cells at exponential growth phase was served as experimental group,however,the culture supernatant with macrophages in 1640 medium was served as control group. The morphological changes of macrophages were observed under microscope on the 7th day and 14 th day,respectively,and the expression levels of the cytokines on surface of macrophages including CD64-M1 and CD163-M2 were detected by flow cytometry in different time points. Results The morphous of macrophages was obviously changed on the 7th day after cultivation in experimental group,with cellular apophysis being increased and cells becoming long and thin,however,the morphous of macrophages was not obviously changed in control group. The proportion of CD64-M1 macrophages in experimental group was significantly lower than that in control group（ P〈0. 05）,however,the proportion of CD64-M2 macrophages in experimental group was significantly higher than that in control group（ P〈0. 05）. On the 14 th day after cultivation the morphous of macrophages was gradually recovered in experimental group,however,the morphous of macrophages was still not obviously changed in control group. Moreover the proportion of CD64-M1 macrophages in experimental group was significantly lower than that in control group（ P〈0. 05）,however,the proportion of CD64-M2 macrophages in experimental group was significantly higher than that in control group（ P〈0. 05）. The ratio of M1 / M0 of CD64-M1 macrophages in experimental group on the 7th day was significantly higher than that on the 14 th day（ P〈0. 05）,however,which in CD163-M2

关 键 词：卵巢肿瘤 SKOV3 巨噬细胞 细胞分化 

分 类 号：R737.31[医药卫生—肿瘤]