IL-8诱导的肿瘤相关巨噬细胞对肝细胞肝癌侵袭转移的影响

摘要

目的:探究外源性IL-8对单核细胞的活化作用及IL-8活化后的肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)对肝细胞肝癌(hepatocellular carcinoma,HCC)侵袭转移的影响.方法:外源性IL-8刺激THP-1细胞后检测M1、M2型TAMs的比例.IL-8活化的TAMs与HCC细胞共孵育,利用RT-PCR和Western blot法检测TAMs对HCC细胞上皮间质转化(epithelial-mes?enchymal transition,EMT)的影响;利用划痕修复实验和Transwell侵袭实验探究TAMs对HCC细胞侵袭转移的影响;并在100例HCC样本中进行组织水平验证.结果:外源性IL-8可诱导THP-1细胞的表型向M2型TAMs方向分化.IL-8体外诱导的TAMs可以反作用于HCC,促使其发生EMT,进而促进其侵袭转移.在HCC组织中证实IL-8与TAMs浸润呈明显的正相关(r=0.22,P<0.05),同时浸润的TAMs与神经细胞型钙黏蛋白(N-cadherin)的表达量呈正相关(r=0.20,P<0.05).结论:在HCC中,IL-8可以趋化TAMs浸润并促进其活化,活化后的TAMs可促进HCC细胞EMT和侵袭转移.%Objective:To study the effects of IL-8 on the polarization of monocytes and the effects of IL-8-induced tumor-associated macrophages(TAMs)on the invasion and metastasis of hepatocellular carcinoma(HCC).Methods:After exogenous IL-8 stimulation of THP-1 cells for 72h,the percentages of M1 and M2 TAMs were examined.RT-PCR and Western blot assays were used to study epitheli-al-mesenchymal transition(EMT),and wound-healing and transwell assays were preformed to study the invasion potential of HCC cells after co-culturing with TAMs and HCC cell lines in vitro.Lastly,100 cases of HCC tissue samples were used to validate the correlation among TAM numbers,IL-8,and EMT features of HCC cells via immunohistochemistry(IHC)staining methods.Results:Exogenous IL-8 induced significant M2 polarization of TAMs in THP-1 cells.TAMs further promoted EMT in HCC and enhanced the invasion potential of HCC in vitro.Finally,significant positive correlations among the numbers of TAMs,IL-8 expression,and N-cadherin expression were identified in primary HCC tissue samples(r=0.22,r=0.20,P<0.05).Conclusions:IL-8 locally attracted and activated TAMs,and promot-ed M2 polarization of TAMs,which further promoted the EMT and invasion potential of HCC cells both in vitro and in vivo.