摘要
目的探讨CCL20-CCR6-Th17轴在肝细胞癌血管浸润转移中的作用。方法采用SYBRGreen实时定量PCR测定人肝细胞系L-02、肝细胞癌细胞系Hep3B、Huh7和HepG2中CCL20mRNA的表达，采用酶联免疫吸附试验（ELISA）检测上清中CCL20蛋白的分泌，Transwell迁移实验检测肝细胞癌细胞系对人外周血单个核细胞的趋化作用。采用ELISA定量检测93例肝细胞癌患者（转移组51例，无转移组42例）术前外周血中白细胞介素10L（IL-1α）、IL-1β、IL-6、IL-8、IL-10、IL-17、IL-23、γ-干扰素、肿瘤坏死因子α和CCL20的水平，采用SYBRGreenRT—PCR检测41例肝细胞癌癌组织及其相应的癌旁组织中CCL20和CCR6的表达水平，免疫组织化学染色检测肝细胞癌及其对应的癌旁组织和正常人供肝组织中CCL20的表达水平。结果人肝细胞癌细胞系中CCL20的表达水平高于正常肝细胞，对外周血表达CCR6的T细胞具有趋化作用。93例肝细胞癌患者血清中CCL20蛋白的表达水平为（38．2±28．4）pg／ml，高于肝血管瘤患者[（7．8±17．8）pg／ml，P〈0．01]。肝细胞癌患者血清中CCL20蛋白的表达水平与肿瘤直径呈正相关（r=0．32，P=0．0018）。41例肝细胞癌组织CCL20mRNA的表达水平高于癌旁组织（P〈0．05），CCL20蛋白主要在肝细胞癌细胞质中表达，某些浸润的免疫细胞也有一定程度表达。多因素分析结果显示，血清中IL-17和CCL20水平是影响肝细胞癌患者发生血管浸润转移的独立影响因素（均P〈0．05）。41例肝细胞癌患者癌组织及对应的癌旁组织中CCL20mRNA的表达水平与肝细胞癌发生血管浸润转移均无关（均P〉0．05）。在发生血管浸润转移患者的癌组织及其对应的癌旁组织中CCR6mRNA的表达水平分别为5．75（1．79，19．13）和7．99（4．49，19．54），均高于未发生血管浸润转移的患者[分别为1．69（0．76，2．87）和3．58（1．84，4．32）]，差异均有�
Objective To investigate the role of CCL20/CCR6/Th17 axis in vascular invasion and metastasis of primary hepatocellular carcinoma （HCC）. Methods Expression levels of CCL20 mRNA in the normal human liver cell line L-02, and human hepatocellular carcinoma cell lines Hep3B, Huh7 and HepG2 were quantified by using SYBR green real time PCR. CCL20 secretions from these cell lines were quantified by using ELISA. The chemotactic effect of HCC cell line Hep3B on human peripheral blood mononuclear cells was determined by using transwell chemotaxis assay. Pre-therapy serum levels of IL-1α, IL-1β, IL-6, IL-8, IL-10, IL-17, IL-23, IFN-γ, TNF-α and CCL20 in 93 patients with HCC were measured by using 9-plex array and ELISA. All the patients were chronic hepatitis B virus associated HCC, and 51 cases were those with vascular invasion and metastasis （ metastasis group） and 42 cases were not （ non-metastasis group）. CCI-20 and CCR6 mRNA expressions in the HCC and tumor-adjacent tissues were determined by using SYBR Green real time PCR in 41 patients, among them, 20 cases were from the group of patients with metastasis and 21 cases were from the group of patients without metastasis. The CCL20 expression was further determined by immunohistochemistry. Results The HCC cell lines expressed and secreted higher amount of CCL20, which effectively recruited CCR6＋ T cells. Pre-therapy serum levels of CCL20 in 93 HCC patients were （38.2±28. g） pg/ml, significantly increased than those with benign hepatic hemangiomas [ （7.8± 17.8）pg/ml, P 〈 0.01 ]. In addition, the serum levels of CCL20 were positively correlated with the tumor diameters in HCC patients （ r = 0.32, P = 0. 0018 ）. CCL20 was dominantly expressed in the cytoplasm in HCC cells, and it was also expressed by some infiltrating immune cells. The mRNA expression levels of CCL20 of the tumor tissues were significantly higher than that in the tumor-adjacent tissues （ P 〈 0.05）. Multivariate logistic regression analysis showed that serum levels o
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2015年第1期5-10,共6页
Chinese Journal of Oncology
基金
国家重点基础研究发展规划项目（2011CB504205）
国家自然科学基金（30973387）
