JNK通路对M2巨噬细胞极化及其肿瘤效应的影响

Effects of JNK Pathway on Polarization and Pro-tumor Function of M2 Macrophage

探究了JNK通路对M2巨噬细胞极化及M2介导的促肿瘤效应的影响。构建单核细胞THP1来源M2巨噬细胞模型（THP1-M2）,将细胞分为3组：用PMA诱导的未活化巨噬细胞组（M0）,用PMA、IL-4处理及阴性干扰（DMSO）的M2型巨噬细胞组（M2）,用特异性抑制剂阻断JNK通路的M2型巨噬细胞组（M2-JNKI）。实时荧光定量PCR检测M2表型marker基因的表达;免疫蛋白印迹法检测M2表型marker蛋白水平;细胞划痕试验检测巨噬细胞迁移能力;流式细胞数检测786O及OSRC2凋亡。结果与THP1-M2组相比,阻断JNK通路的M2组M2表型marker表达明显下降,同时其细胞迁移能力也呈下降趋势。且阻断JNK通路后,M2巨噬细胞抑制肾癌细胞凋亡的能力减弱。结果表明,抑制JNK通路后,M2巨噬细胞极化状态受损,其促肿瘤效应可转变为抗肿瘤效应。

The present study investigates the effect of JNK pathway on the polarization of M2 status as well as pro-tumor function mediated by M2. THP1 derived M2 macrophage（ THP1-M2） model was established. The cells were divided into 3 groups： the PMA pretreated unpolarized macrophage（ M0）,the PMA and IL-4 induced M2 macrophage with DMSO（ negative control） treated（ M2）,the JNK inhibitor treated M2 macrophage（ M2-JNK I）. Furthermore,the M2 associated markers Arginase1（ Arg1）,mannose receptor C-type 1（ Mrc1） were analyzed by Q-PCR,the protein level of Arg1 and Mrc1 were detected by Western blot,the migration ability of macrophages was tested by Wound Healing,the apoptosis of 786 O and OSRC2 were analyzed by flow cytometry.Compared with the THP1-M2,THP1-M2-JNK I group showed decreased expression of Arg1 and Mrc1,and migration ability was impaired. What＇s more,block of JNK pathway inhibited the pro-tumor function of M2 on786 O and OSCR2. Taken together,the results suggest that inhibition of JNK pathway regulates M2 polarization and its pro-tumor effects.