白介素-35在M2型巨噬细胞中的表达及其对前列腺癌细胞的增殖、侵袭和上皮间质转化的影响

The expression levels of interleukin-35 in M2 subtype macrophages and its effects on the proliferation invasion and epithelial-mesenchymal transition of prostate cancer cells

目的探讨白介素(IL)-35在巨噬细胞中的表达及其对前列腺癌(PCa)细胞增殖、侵袭和上皮间质转化(EMT)的影响和机制。方法检测巨噬细胞标记分子CD68、M1型标记分子肿瘤坏死因子-α(TNF-α)、IL-1β和M2型标记分子CD206、IL-10在人单核/巨噬细胞系THP-1细胞诱导分化后的M0、M1与M2型巨噬细胞中的表达。使用不同类型的巨噬细胞-源条件培养液(CM)处理PCa细胞系PC-3,并将其分为:M0-CM组、M1-CM组、M2-CM组和预先使用IL-35中和抗体(IL-35 NA)干预的M2-CM处理组(M2-CM+IL-35 NA组)。检测各组中PC-3细胞的增殖活力、侵袭、EMT,以及细胞内JAK/STAT信号通路p-JAK2/JAK2与p-STAT3/STAT3比值的差异。结果与未刺激的THP-1细胞比较,M0、M1和M2型巨噬细胞中CD68 mRNA表达升高(P<0.01);与M0型巨噬细胞比较,TNF-α、IL-1βmRNA在M1型巨噬细胞中升高(P<0.01),而CD206、IL-10 mRNA在M2型巨噬细胞中升高(P<0.01);与M0-CM组比较,M2-CM组PC-3细胞增殖活力、侵袭、EMT和细胞内p-JAK2/JAK2、p-STAT3/STAT3比值升高(P<0.05),M2-CM+IL-35 NA组PC-3细胞的增殖活力、侵袭、EMT和细胞内p-JAK2/JAK2、p-STAT3/STAT3比值均显著降低(P<0.05),M1-CM组PC-3细胞的增殖活力、侵袭、EMT和细胞内p-JAK2/JAK2、p-STAT3/STAT3比值均无显著改变(P>0.05);与M2-CM组比较,M1-CM组和M2-CM+IL-35 NA组增殖活力、侵袭、EMT和细胞内p-JAK2/JAK2、p-STAT3/STAT3比值降低(P<0.05)。结论M2型巨噬细胞可通过分泌IL-35来促进PC-3细胞的增殖、侵袭和EMT,这可能与IL-35能上调JAK2/STAT3信号通路的活化相关。

Objective To explore the expression levels of interleukin-35(IL-35)in macrophages and its effects and mechanisms on the proliferation,invasion,and epithelial-mesenchymal transition(EMT)of prostate cancer(PCa)cells.Methods The expression levels of macrophage marker CD68,M1 subtype marker tumor necrosis factor-α(TNF-α)and IL-1β,and M2 subtype marker CD206 and IL-10 in M0,M1 and M2 macrophages after induction and differentiation of human mononuclear/macrophage line THP-1 cells were detected.PCa cell line PC-3 cells were treated with different types of macrophages-sources conditioned medium(CM),and were divided into M0-CM group,M1-CM group,M2-CM group,and M2-CM group(M2-CM+IL-35 NA group)which was pre-interfered with IL-35 neutralizing antibody(IL-35 NA).The proliferation,invasion and EMT of PC-3 cells and the p-JAK2/JAK2 ratio and p-STAT3/STAT3 ratio of JAK/STAT signaling pathway were detected.Results Compared with unstimulated THP-1 cells,CD68 mRNA expression levels were increased in M0,M1 and M2 subtype macrophages(P<0.01).Compared with M0 subtype macrophages,TNF-αand IL-1βmRNA were increased in M1 subtype macrophages(P<0.01),while CD206 and IL-10 mRNA were increased in M2 subtype macrophages(P<0.01).Compared with M0-CM group,the proliferation,invasion,EMT,intracellular p-JAK2/JAK2 and p-STAT3/STAT3 ratios of PC-3 cells were increased in M2-CM group(P<0.05).In M2-CM+IL-35 NA group,the proliferation,invasion,EMT,intracellular p-JAK2/JAK2 and p-STAT3/STAT3 ratios of PC-3 cells were significantly decreased(P<0.05).In M1-CM group,the proliferation,invasion,EMT,intracellular p-JAK2/JAK2 and p-STAT3/STAT3 ratios of PC-3 cells had no significantly difference(P>0.05).Compared with M2-CM group,the proliferation,invasion,EMT,intracellular p-JAK2/JAK2 and p-STAT3/STAT3 ratios were decreased in M1-CM group and M2-CM+IL-35 NA group(P<0.05).Conclusions M2 subtype macrophages can promote the proliferation,invasion and EMT of PC-3 cells by secreting IL-35,which may be related to the activation of JAK2/STAT3 signaling pathway by IL-35.