姜黄素干预乳腺癌细胞诱导的巨噬细胞极化效应

Role of Curcumin in Macraphage Polarization Induced by Breast Cancer Cells

目的探讨姜黄素对乳腺癌细胞诱导的巨噬细胞极化的干预效应。方法采用Transwell共培养体系,将小鼠乳腺癌细胞4T1和小鼠巨噬细胞RAM264.7体外共培养5d。实验组加入不同浓度姜黄素处理。流式细胞术检测共培养后巨噬细胞CD163和F4/80的表达;将共培养后的巨噬细胞无血清培养过夜,留取上清,明胶酶谱(Zymography)法检测细胞分泌金属基质蛋白酶-9(metal matrix protease-9,MMP-9)水平;收集共培养后的巨噬细胞,Transwell检测共培养后巨噬细胞的迁移能力。建立小鼠乳腺癌原位肿瘤模型,姜黄素灌胃给药,监测肿瘤生长,记录小鼠荷瘤生存期;免疫荧光检测移植瘤中CD163^+/F4/80^+巨噬细胞的比例。结果体外实验显示,姜黄素能有效干预乳腺癌诱导的巨噬细胞CD163和F4/80表达以及MMP-9分泌,且明显减弱共培养巨噬细胞的体外迁移能力。姜黄素体内给药,虽不能明显抑制肿瘤原位生长,但是可以明显延长小鼠荷瘤生存期,并降低移植瘤中CD163^+/F4/80^+巨噬细胞的比例。结论姜黄素能干预乳腺癌细胞诱导的巨噬细胞极化,发挥抗瘤效应。

Objective To investigate the role of curcumin in macraphage polarization induced by breast cancer cells.Methods By using Transwell co-culture technique,murine breast cancer 4T1 cells were incubated with RAW264.7 macrophages for 5 days in vitro.Cells in the experiment group were pretreated with curcumin.Flow cytometry was used to detect the CD163 and F4/80 expression on macraphages and zymography to analyze the activity of MMP-9 secreted by co-cultured macrophages after treatment in the absence of serum for a night.Migration of macrophages towards breast cancer cell media was investigated by Transwell assay.Additionally,curcumin was orally administrated to mice bearing orthotopic transplantation tumors.Tumor growth was observed as well as overall survival.Furthermore,the CD163+/F4/80+macrophages in orthotopic transplantation tumor was measured by immunofluorescence.Results In vitro experiments showed that curcumin could dramatically suppress the expression of CD163 and F4/80 on macrophages,the secretion of MMP-9 and the migration of macrophages.Furthermore,while the tumor growth was not significantly inhibited by curcumin,in vivo delivery of curcumin could prolong the overall survival of tumor-bearing mice and downregulate the proportion of CD163+/F4/80+macrophages in orthotopic transplantation tumors.Conclusion Curcumin could inhibit the macraphage polarization induced by breast cancer cells.