结缔组织生长因子在人肝癌中过表达，促进细胞侵袭和生长。
Connective tissue growth factor is overexpressed in human hepatocellular carcinoma and promotes cell invasion and growth.
机构信息
Department of Hepatic-Biliary-Pancreatic Medicine, First Hospital, Jilin University, Changchun 130021, Jilin Province, China.
出版信息
World J Gastroenterol. 2012 Dec 21;18(47):7070-8. doi: 10.3748/wjg.v18.i47.7070.
AIM
To determine the expression characteristics of connective tissue growth factor (CTGF/CCN2) in human hepatocellular carcinoma (HCC) in histology and to elucidate the roles of CCN2 on hepatoma cell cycle progression and metastasis in vitro.
METHODS
Liver samples from 36 patients (who underwent hepatic resection for the first HCC between 2006 and 2011) and 6 normal individuals were examined for transforming growth factor β1 (TGF-β1) or CCN2 mRNA by in situ hybridization. Computer image analysis was performed to measure integrated optimal density of CCN2 mRNA-positive cells in carcinoma foci and the surrounding stroma. Fibroblast-specific protein-1 (FSP-1) and E-cadherin were examined to evaluate the process of epithelial to mesenchymal transition, α-smooth muscle actin and FSP-1 were detected to identify hepatic stellate cells, and CD34 was measured to evaluate the extent of vascularization in liver tissues by immunohistochemical staining. CCN2 was assessed for its stimulation of HepG2 cell migration and invasion using commercial kits while flow cytometry was used to determine CCN2 effects on HepG2 cell-cycle.
RESULTS
In situ hybridization analysis showed that TGF-β1 mRNA was mainly detected in connective tissues and vasculature around carcinoma foci. In comparison to normal controls, CCN2 mRNA was enhanced 1.9-fold in carcinoma foci (12.36 ± 6.08 vs 6.42 ± 2.35) or 9.4-fold in the surrounding stroma (60.27 ± 28.71 vs 6.42 ± 2.35), with concomitant expression of CCN2 and TGF-β1 mRNA in those areas. Epithelial-mesenchymal transition phenotype related with CCN2 was detected in 12/36 (33.3%) of HCC liver samples at the edges between carcinoma foci and vasculature. Incubation of HepG2 cells with CCN2 (100 ng/mL) resulted in more of the cells transitioning into S phase (23.85 ± 2.35 vs 10.94 ± 0.23), and induced a significant migratory (4.0-fold) and invasive (5.7-fold) effect. TGF-β1-induced cell invasion was abrogated by a neutralizing CCN2 antibody showing that CCN2 is a downstream mediator of TGF-β1-induced hepatoma cell invasion.
CONCLUSION
These data support a role for CCN2 in the growth and metastasis of HCC and highlight CCN2 as a potential novel therapeutic target.
目的
确定结缔组织生长因子（CTGF/CCN2）在人肝细胞癌（HCC）组织学中的表达特征，并阐明 CCN2 在体外肝癌细胞周期进展和转移中的作用。
方法
对 36 例（2006 年至 2011 年间首次接受肝切除术治疗 HCC）患者和 6 例正常个体的肝组织样本进行转化生长因子β1（TGF-β1）或 CCN2mRNA 的原位杂交检测。采用计算机图像分析方法测量癌灶和周围基质中 CCN2mRNA 阳性细胞的整合最佳密度。检测纤维母细胞特异性蛋白-1（FSP-1）和 E-钙黏蛋白以评估上皮间质转化过程，检测α-平滑肌肌动蛋白和 FSP-1 以鉴定肝星状细胞，并用免疫组织化学染色法测量 CD34 以评估肝组织中的血管化程度。使用商业试剂盒评估 CCN2 对 HepG2 细胞迁移和侵袭的刺激作用，同时使用流式细胞术确定 CCN2 对 HepG2 细胞周期的影响。
结果
原位杂交分析显示，TGF-β1mRNA 主要在癌灶周围的结缔组织和血管中检测到。与正常对照组相比，癌灶中 CCN2mRNA 增强 1.9 倍（12.36±6.08 vs 6.42±2.35）或周围基质中增强 9.4 倍（60.27±28.71 vs 6.42±2.35），同时这些区域也表达 CCN2 和 TGF-β1mRNA。在 36 例 HCC 肝组织样本中（33.3%），在癌灶和血管之间的边缘处检测到与 CCN2 相关的上皮-间质转化表型。用 CCN2（100ng/ml）孵育 HepG2 细胞会导致更多的细胞进入 S 期（23.85±2.35 vs 10.94±0.23），并诱导显著的迁移（4.0 倍）和侵袭（5.7 倍）作用。用中和 CCN2 抗体阻断 TGF-β1 诱导的细胞侵袭表明，CCN2 是 TGF-β1 诱导肝癌细胞侵袭的下游介质。
结论
这些数据支持 CCN2 在 HCC 生长和转移中的作用，并强调 CCN2 是一种潜在的新型治疗靶点。
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