上皮间质转化标志物E-cadherin和Vimentin在原发性肝癌中的表达及其临床意义

Expression and roles of the epithelial mesenchymal transition markers E-cadherin and vimentin in hepatocellular carcinoma

目的检测上皮间质转化（EMT）标志物E-cadherin和Vimentin在原发性肝癌及相邻正常组织中的蛋白表达水平，探讨E-cadherin和Vimentin表达差异与肝癌转移相关胜及预测临床预后意义。方法收集30例原发性肝癌患者肝癌组织及相邻正常组织，Westernblot荧光发光法定量检测E-cadherin，Vimentin蛋白在肝癌组织及相邻正常组织的表达，分析二者的蛋白表达相互关系及其与临床病理特征的相关性;同时采用免疫组织化学法检测Twist，Vimentin，E—cadherin在肝癌组织及相邻正常组织中的表达。数据统计采用配对t检验，Mann-Whitney U检验，spearman等级相关分析。结果E-cadherin蛋白在原发性肝癌中肝癌组织较相邻正常组织表达显著降低，相对表达量为0．082±0．063对比0．226±0．215，差异具有统计学意义，t=-4．050，P〈0．01；其在门静脉癌栓患者肝癌组织中表达较无门静脉癌栓者降低妒=0．001），其在TNMm期患者肝癌组织中表达较TNM I／II期表达降低（p=0．003）；Vimentin蛋白在原发性肝癌中癌组织表达较相邻正常组织表达增高（p=0．002），Vimentin蛋白表达与E-cadherin蛋白表达呈负相关（r=-0．509，P=0．004），Vimentin蛋白表达与门静脉癌栓（P〈0．01）、TNM分期妒〈0．01）、Milan标准（P=0．005）存在相关性；免疫组织化学结果显示E—cadherin在癌组织中几乎不表达，在相邻正常组织细胞膜上高表达，Vimentin和Twist均在肝癌组织细胞中高表达，而在相邻正常组织不表达。结论EMT标志物E—cadherin和Vimentin蛋白在原发性肝癌中的表达与患者门静脉癌栓、TNM分期等临床特征密切相关，E—cadherin和Vimentin可作为预测肝癌转移复发及评估临床预后的有效指标。

Objective To determine the differential protein expressions of epithelial mesenchymal transition （EMT） markers E-cadherin and vimentin in hepatocellular carcinorma （HCC） and to investigate their correlation to the molecular mechanisms of metastasis to explore their potential utility as prognostic indicators of HCC. Methods Tumor tissues and patient-matched adjacent non-tumor tissues were collected from individuals diagnosed with HCC. E-cadherin and vimentin protein expressions in the tissue specimens were quantified by western blot with densitometry of fluorescence emission and comparatively analyzed to determine the associations with molecular and clinical features. The expressions of E-cadherin and vimentin, as well as the other EMT-related protein Twist, were also detected in the tissue specimens by immunohistochemistry. Statistical analyses were carried out by paired-samples t-test, Mann-Whitney test, and Spearman rank correlation analysis. Results E-cadherin expression was significantly lower in tumor tissues （0.082 ± 0.063 vs. adjacent non-tumor tissues： 0.226 ± 0.215, t = -4.050, P 〈 0.01）, lower in patients with portal vein tumor thrombus （vs. non-thrombic HCC patients, P = 0.001）, and correlated with TNM stage （III/ IV 〉 I/II, P = 0.003）. Vimentin expression was significantly higher in tumor tissues （vs. adjacent non-tumor tissues, P = 0.002）, negatively correlated with E-cadherin expression （t = -0.509, P = 0.004）, and closely associated with some clinical parameters, such as portal vein tumor thrombus （P 〈 0.01）, TNM stage （P 〈 0.01）, and Milan criteria （P = 0.005）. Immunohistochemistry showed that E-cadherin expression was very weak in tumors but very strong in the cell membranes of non-tttrnor tissues, and that vimentin and Twist expressions were strong in tumors but undetectable in non-tumor tissue. Conclusion Expression levels of the EMT markers E-cadherin and vimentin in HCC are related to clinical parameters, including portal vein tumor thrombus and TNM stage, and may represent useful prognostic markers of metastasis.