Correlation between inflammation factors and hepatic cancer stem cell markers during hepatocarcinogenesis
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摘要: 目的：探讨 Kupffer 细胞相关炎症因子在大鼠肝癌诱导过程中的动态变化与肝癌干细胞标志的相关性。方法选取雄性 SD 大鼠35只，分为正常对照组（n ＝5）和诱导模型组（n ＝30）。诱导模型组大鼠饮水中加入二乙基亚硝胺溶液，正常对照组给予普通饮水。在诱癌过程中每隔4周取材5只大鼠肝脏，运用免疫组化法测定 Kupffer 细胞 ED2抗体在肝组织的表达和定量 PCR 方法检测肝组织中 Kupffer 细胞释放的炎症因子及肝癌干细胞标志物的基因表达。并采用 Pearson 检验分析相关炎症因子与肝癌干细胞标志物之间相关性。结果成功建立大鼠肝癌诱导模型，在24周时诱导模型组大鼠出现肝脏肿瘤；ED2抗体在诱癌过程中呈明显升高的趋势（P ＜0．001）；PCR 结果提示，Kupffer 细胞分泌的相关炎症因子 IL-6、MCP-1、TGF-β、TNF-α在诱癌过程中逐渐升高（P ＜0．05），同时，肝癌干细胞标志物 CD90在诱癌过程中呈明显升高趋势（P ＜0．001）。Pearson相关性检验提示：IL-6、MCP-1、TGF-β的表达变化与 CD90呈明显正相关（P ＜0．001）。结论Kupffer 细胞释放的炎症因子 IL-6、MCP-1、TGF-β的表达升高与 CD90上调呈明显正相关，进一步明确了 Kupffer 细胞促进肝癌发生的重要作用。Abstract: Objective To investigate the correlation between inflammation factors of Kupffer cells and hepatic cancer stem cell markers.Methods 35 male SD rats were divided randomly into two groups:normal group (5 rats)and diethylnitrosamine (DEN)-induced groups (30 rats).DEN-in-duced groups were given DEN solution in drinking water,while normal group was given normal water. 5 rats of DEN-induced group were sacrificed every 4 weeks,then immunohistochemisty was taken to e-valuate the ED2 expression of Kupffer cells,and the expressions of inflammation factors and hepatic cancer stem cell markers were measured by rt-PCR.Correlation between inflammation factors and he-patic cancer stem cell markers was evaluated by Pearson analysis.Results The HCC model was suc-cessfully established at 24 weeks.Expressions of ED2 gradually increased (P <0.001 ).The rt-PCR results indicated that the expressions of IL-6、MCP-1、TGF-βand TNF-αgradually increased during hepatocarcinogenesis (P <0 .0 5 ),while CD90 expressed a significant increasing in the process (P <0 .00 1 ).Pearson analysis showed CD90 was positively correlated with IL -6 、MCP -1 and TGF -β(P <0.001).Conclusion There is a significant correlation between the increase expressions of IL-6、MCP-1、TGF-βand the up-regulation of CD90 ,which can demonstrate the essential role of Kupffer cells during hepatocarcinogenesis.
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