目的探讨卵巢癌组织中M2型巨噬细胞的分布及其对卵巢癌侵袭转移相关基质金属蛋白酶(MMPs)表达的影响。方法免疫组织化学法检测上皮性卵巢癌(34例)、交界性卵巢肿瘤(16例)和卵巢良性肿瘤(18例)组织切片中CD68、CD206和MMP-9的表达,比较卵巢癌样本中阳性表达细胞数与患者年龄、病理类型、FIGO分期、组织学分化和淋巴结转移的关系。并利用流式细胞术比较其中12例上皮性卵巢癌和11例卵巢良性肿瘤新鲜组织中CD68+~CD206+~细胞比例。实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印记(Western blot)方法分别评价卵巢癌和卵巢良性肿瘤组织内MMP-2、MMP-9和MMP-10水平。结果卵巢癌组织中CD68+~、CD206+~和MMP-9+~细胞数量明显高于交界性卵巢肿瘤和卵巢良性肿瘤组织(P<0.05),卵巢癌组织中CD68+~、CD206+~和MMP-9+~细胞数与患者FIGO分期和淋巴结转移均有统计学关系(P<0.05)。卵巢癌组织中CD68+~和CD206+~细胞数量呈正相关关系(r=0.508,P<0.05),且MMP-9+~与CD68+~和CD206+~细胞数量也分别呈正相关(r=0.611,P<0.001;r=0.744,P<0.001)。卵巢癌新鲜组织中CD68+~CD206+~细胞比例明显高于良性卵巢肿瘤(P<0.05),且卵巢癌组织MMP-2、MMP-9和MMP-10 m RNA和蛋白表达水平较卵巢良性肿瘤具有统计学意义(P<0.05)。结论卵巢癌组织中具有较高水平的M2型巨噬细胞,与卵巢癌侵袭转移和临床进展有重要的联系。
Background:Inflammation is often linked with the progress and poor outcome of lung cancer.The understanding of the relationship between tumor-associated macrophages(TAMs) and lung cancer cells involves in the underlying mechanism of inflammatory cytokine production.Toll-like receptors(TLRs) are engaged in promoting the production of pro-inflammatory cytokines and play an important role in tumor immunology.Methods:To investigate the mechanisms by which TAMs influence the production of pro-inflammatory cytokines in lung cancer cells,we established an in vitro coculture system using TAMs and human nonsmall cell lung cancer(NSCLC) cell line SPC-A1.Levels of interleukin(IL)-1β,IL-6 and IL-8 in SPC-A1 were evaluated by RT-PCR and cytometric bead array assay after being cocultured with TAMs.Expression changes of TLRs and TLRs signaling pathway proteins in SPC-A1 were further confirmed by RT-PCR and western blot.The level changes of IL-1β,IL-6 and IL-8 in SPC-A1 were also detected after the stimulation of TLRs agonists.Results:We found that the phenotype markers of TAMs were highly expressed after stimulating human monocyte cell line THP-1 by phorbol-12-myristate-13-acetate(PMA).Higher mRNA and supernate secretion levels of IL-1β,IL-6 and IL-8 were detected in SPC-A1 after being cocultured with TAMs.We also found that TLR1,TLR6 and TLR7 were up-regulated in SPC-A1 in the coculture system with TAMs.Meanwhile,TLRs signaling pathway proteins were also significantly activated.Moreover,pre-treatment with agonist ligands for TLR1,TLR6 and TLR7 could dramatically promote inductions of IL-1β,IL-6 and IL-8.Conclusions:These findings demonstrated that TAMs may enhance IL-1β,IL-6 and IL-8 expressions via TLRs signaling pathway.We conclude that TAMs contribute to maintain the inflammation microenvironment and ultimately promote the development and progression of lung cancer.
