摘要
目的:观察M1型巨噬细胞对人乳腺癌细胞系-7(michigan cancer foundation-7,MCF-7)迁移和侵袭的影响,并对其作用机制进行探讨。方法:以CD44+乳腺癌MCF-7细胞为研究模型,转化人单核细胞系-1(human monocyte cell line,THP-1)为M1型巨噬细胞,培养并收集Ⅰ型巨噬细胞条件培养基(macrophage1-conditioned medium,M1CM),M1CM刺激模型细胞设为M1CM组,同时设立对照组;划痕实验检测细胞的迁移能力;侵袭小室实验考察细胞的侵袭能力;Western blot实验检测各蛋白表达变化;siRNA转染干扰模型细胞Smad1或CD44蛋白表达。结果:THP1经药物诱导后呈圆形、椭圆形、三角形贴壁生长,部分细胞可见伪足和凸起,细胞可吞噬墨汁粒呈蓝黑色,与M0巨噬细胞相比,细胞中M1巨噬细胞标志物CD80和人血清趋化因子配体9(human CXC-chemokine lig-9,CXCL-9)mRNA水平显著升高,差异具有统计学意义(t值分别为14.01和12.22,P值均<0.05)。与对照组相比,M1CM处理24 h显著增强模型细胞迁移和侵袭,差异具有统计学意义(t值分别为23.12和6.73,P值均<0.05)。与CD44小干扰对照(siCON1)相比,M1CM处理24 h明显诱导模型细胞Smad1和CD44蛋白表达增加,差异具有统计学意义(t值分别为6.66和6.25,P值均<0.05)。与Smad1小干扰对照(siCON2)相比,干扰CD44(siCD44)明显抑制M1CM诱导的模型细胞迁移和侵袭,差异具有统计学意义(t值分别为8.17和16.25,P值均<0.05);与siCON1相比,干扰Smad1明显抑制M1CM诱导的模型细胞迁移和侵袭(t值分别为7.38和5.62,P值均<0.05),siCD44明显抑制M1CM诱导的模型细胞Smad1蛋白表达上调(t值分别为14.65和10.02,P值均<0.05),差异均具有统计学意义;与siCON2相比,siSmad1明显抑制M1CM诱导的模型细胞CD44蛋白表达上调,差异具有统计学意义(t值分别为6.65和11.53,P值均<0.05)。结论:M1CM能诱导乳腺癌MCF-7细胞迁移和侵袭能力增强,而该作用可能与CD44-Smad1环路信号增强有关。
Objective To observe the effects of M1 macrophages on the migration and invasion of michigan cancer foundation-7(MCF-7)breast cancer cells,and to explore its signaling mechanism.Methods Human monocyte line-1(THP-1)was transformed into M1 macrophage by using CD44+breast cancer MCF-7 cell as the research model.Macrophage1-conditioned medium(M1CM)was cultured and collected.The model cells stimulated by M1CM were divided into M1CM group,and at the same time the control group was set up.Wound healing experiments test was used to detect the migration ability of cells.Cell invasiveness was tested by invasion chamber experiments.Western blot experiment was used to detect the expression of each protein.Results THP1 was circular,oval,and triangular adherent growth after drug induction.Pseudopodia and bulge can be seen in some cells,and the cells could swallow ink granules in blue and black.Compared with M0 macrophages,the mRNA levels of M1 macrophage marker CD80 and human CXC-chemokine lig-9(CXCL-9)in cells were significantly increased in THP1(t values were 14.01 and 12.22,respectively,both P values<0.05).Compared with the control group,the cell migration and invasion of the model cell were significantly enhanced after 24 hours of M1CM treatment(t values were 23.12 and 6.73,respectively,both P values<0.05).Compared with the CD44 small interference control(siCON1),the expression of Smad1 and CD44 in the model cells was significantly increased after 24 hours of M1CM treatment(t values were 6.66 and 6.25,respectively,both P values<0.05).Compared with Smad1 small interference control(siCON2),the ability of cell migration and invasion induced by M1CM were significantly inhibited by interference with CD44(siCD44)(t values were 8.17 and 16.25,respectively,both P values<0.05).Compared with siCON1,at the same time,interference with Smad1 could also significantly inhibit the cell migration and invasion induced by M1CM(t values were 7.38 and 5.62,respectively,both P values<0.05),and siCD44 significantly inhibited M1CM-induced upregul
作者
边小博
邱芳
韩彦槊
于宏
Bian Xiaobo;Qiu Fang;Han Yanshou;Yu Hong(Department of General Surgery,Shengjing Hospital of China Medical University,Shenyang 110021,China)
出处
《国际免疫学杂志》
CAS
2020年第3期239-244,共6页
International Journal of Immunology
基金
国家自然科学基金青年基金(81600370)。
