胰腺癌细胞Sonic Hedgehog的分泌及对巨噬细胞极化的影响

Secretion of Sonic Hedgehog in pancreatic cancer cells and its effect on macrophage polarization

目的探讨胰腺肿瘤细胞中Shh(Sonic Hedgehog)的分泌情况及对巨噬细胞极化的影响。方法应用GEPIA 2数据库分析Shh在胰腺癌组织与癌旁正常组织中的表达差异;体外培养正常导管上皮细胞株HPDE6-C7和胰腺癌细胞株PANC-1和CFPAC-1,通过酶联免疫吸附技术(ELISA)检测其上清液中Shh的含量,应用胰腺癌细胞上清液作用于巨噬细胞株RAW264.7,以及单纯性应用重组Shh蛋白作用于巨噬细胞,通过细胞免疫荧光染色检测M2型巨噬细胞标志物精氨酸酶-1(Arg-1)、干扰素调节因子4(IRF4)的表达水平。结果GEPIA 2数据库显示,与癌旁正常组织相比,Shh在胰腺癌组织中的表达明显升高(P<0.05);同时ELISA分析显示,与正常导管上皮细胞HPDE6-C7相比,胰腺癌细胞PANC-1和CFPAC-1中Shh的表达水平明显增高(P<0.05)。应用胰腺癌细胞的上清液作用于巨噬细胞RAW264.7后,其Arg1的表达明显增加(P<0.05)。同样,应用重组Shh作用后可增加巨噬细胞中Arg-1的表达(P<0.05);胰腺癌细胞上清液或Shh处理巨噬细胞后,与M2型极化相关的转录因子IRF4的表达明显增加(P<0.05)。结论胰腺癌细胞通过释放Shh提高巨噬细胞中IRF4的表达,进而诱导巨噬细胞向M2型极化。

Objective To investigate the release of Sonic Hedgehog(Shh)in pancreatic cancer cells and its effect on the polarization of macrophages.Methods The GEPIA 2 database was used to analyze the expression difference of Shh in pancreatic cancer tissues and adjacent normal tissues.ELISA was used to detect the release of Shh in the supernatant of normal ductal epithelial cells(HPDE6-C7)and pancreatic cancer cells(PANC-1 and CFPAC-1 cells)in vitro.In addition,in macrophages(RAW264.7)treated with the supernatant of pancreatic cancer cells orrecombinant Shh protein,the expression levels of M2 macrophage marker arginase-1(Arg-1)and interferon regulatory factor 4(IRF4)were detected by cell immunofluorescence staining.Results:The GEPIA 2 database showed that Shh expression in pancreatic cancer tissues was significantly higher than that in normal tissues(P<0.05).ELISA analysis showed that compared with HPDE-C7 cells,the expression levels of Shh in PANC-1 and CFPAC-1 cells were significantly higher(P<0.05).After treatment with the supernatant of pancreatic cancer cells in cultured macrophages,the expression of M2 macrophage marker Arg-1 was significantly increased(P<0.05).Besides,treatment with recombinant Shh increasedArg-1 expression in RAW264.7.Further analysis showed that the supernatant of pancreatic cancer cancer cells or Shh increased the expression of IRF4(P<0.05).Conclusion Shh released from pancreatic cancer cells induces M2 macrophage polarization by enhancing IRF4 expression.