肝细胞肝癌NTS的表达与炎症微环境形成和肿瘤上皮间质转化及预后的相关性研究*

摘要

Objective:This work aims determine the expression of the neurotensin (NTS) gene in hepatocellular carcinoma (HCC) subgrouping using immunohistochemical staining (IHC) as well as to evaluate the correlation between the activation of NTS/IL-8 pathway in HCC and inflammatory response in microenvironment and epithelial mesenchymal transition (EMT) in cancer and in the prognosis of patients. Methods:Tumor tissues and corresponding adjacent normal tissue were collected from 64 cases of HCC patients. The expression levels of NTS protein and multiple inflammation and EMT-related proteins, including IL-8, VEGF, MMP9, CD68, E-Cadherin,β-Catenin, and Vimentin, were examined in 64 cases of paraffin-embedded HCC tissues using the immunohistochemistry (IHC) staining method. The clinical outcome and overall survival (OS) among 64 cases of HCC patients were compared. Results:We found that the frequency of NTS-expressing tissues among all HCC samples was 17.19%(11/64). Significantly increased IL-8 protein was confirmed in 90.91%of NTS+HCC samples and was positively correlated with the levels of NTS protein in cancer tissues (P=0.036), which implied the dysfunctional activation of NTS/IL-8 pathway in HCC. The levels of VEGF and MMP9 were significantly correlated with the co-expression of NTS and IL-8 in HCC. Evident features of EMT, including decreased membrane expression of E-Cadherin and increased accumulation of cytoplasmicβ-Catemin and Vimentin, were found in NTS+IL-8+samples. The co-expression of NTS and IL-8 in cancer was significantly correlated with the clinical outcomes of patients, as the mortality rate of NTS+IL-8+HCC patients is 2.5-fold higher than that of others after surgery (P=0.022).Accordingly, the OS of NTS+IL-8+HCC patients significantly decreased (24.65±4.45 m vs. 75.79±16.32 m, P=0.013), and these patients are at a higher risk of death at an expected hazard ratio (HR) of 3.457. Conclusion:The NTS/IL-8 pathway is dysfunctionally activated in a subgroup of HCC samples. Highly expressed NTS is associated with increased inflammatory response in microenvironment, enhanced EMT in cancer, and worse prognosis in HCC patients.%目的：利用免疫组织化学染色法（IHC）检测肝细胞肝癌（HCC）中神经降压素（NTS）的表达，并探讨HCC中NTS/IL-8通路的激活与炎性微环境形成和肿瘤上皮间质转化（EMT）及临床预后的关系。方法：收集本院2007年11月至2009年7月期间进行部分肝切除术后确诊为肝细胞癌（HCC）、随访资料完整的肝癌患者64例。采用IHC法检测肝癌及相应癌旁正常组织中NTS，多种炎症因子和EMT相关蛋白的表达情况，包括IL-8、VEGF、MMP-9、CD68、E-cadherin，β-Catenin和Vimentin的表达水平，并比较不同NTS表达对HCC患者总生存时间（OS）的影响。结果：NTS在HCC中表达率为17.19%（11/64），NTS阳性HCC标本中IL-8阳性率较NTS阴性标本明显增加（90.91%vs.41.17%），二者表达呈正相关（R2=0.298，P=0.006）。NTS和IL-8双阳性（NTS+IL-8+）HCC标本中，VEGF和MMP-9表达显著增加。同时，NTS+IL-8+HCC细胞EMT特征明显，表现为E-Cadherin表达降低和Vi-mentin表达升高。在肿瘤中NTS和IL-8共表达与患者的临床疗效呈正相关，术后NTS+IL-8+HCC患者的死亡率比非NTS和IL-8共表达的患者高2.5倍（P=0.022）。NTS+IL-8+HCC患者的OS显著缩短[（24.65±4.45）个月vs.（75.79±16.32）个月，P=0.013）]，而死亡风险明显升高，其预期危险比（HR）为3.457。结论：在部分HCC中存在NTS/IL-8炎症通路的异常活化，NTS的高表达与炎症微环境形成和肿瘤EMT与肝癌患者预后较差密切相关。