NET-1蛋白在肝癌中的表达和意义

Expression and significance of NET-1 protine in hepatocellular carcinoma

目的研究NET-1基因在肝癌中的表达和意义,分析其与临床病理因素的相关性。方法Western blot法检测8例肝癌组织中NET-1的表达,免疫荧光细胞化学法检测肝癌SMMC-7721细胞中NET-1蛋白的表达,免疫组化法检测130例肝癌石蜡包埋组织中NET-1蛋白的表达。结果Western blot法检测显示,8例肝癌组织匀浆中均有NET-1的表达。免疫荧光细胞化学激光共聚焦观察显示,NET-1在SMMC-7721细胞浆中表达呈不规则颗粒状,分布在高尔基体附近。免疫组化结果显示,肝癌组织中NET-1蛋白检出率为96.9%(126/130),其表达强度与癌细胞学类型密切相关;在透明细胞型、多形细胞型和肉瘤样型中,NET-1蛋白检出率和阳性强度显著高于肝细胞型(P< 0.05)。NET-1基因与癌病理分级、临床分期和癌伴肝炎肝硬化呈等级正相关,而与癌伴片块状坏死呈负相关(P均<0.05),与患者AFP水平、肿瘤大小及肿瘤生长方式无明显相关。结论在肝癌中,NET-1表达于胞浆,定位于近高尔基体处。NET-1蛋白表达可能促进肝癌细胞失控性增生和失分化。

Objective This study was designed to investigate the expression and significance of NET-1 in hepatocellular carcinoma （HCC） and analyze the relationship between NET-1 gene expression and clinicopathologic factors in HCC. Methods NET-1 gene protein expression was detected by Western blot, fluorescence immunocytochemistry, confocal laser scanning microscopy and immunohistochemistry in 8 cases of HCC tissues, human hepatoma cell line SMMC-7721, and paraffin-embeded sections from 130 cases of HCC. Results NET-1 gene protein expressed in 8 cases of HCC tissues by Western blot. The NET-1 gene protein positively located in the cytoplasm as irregular granules near Golgi apparatus in SMMC-7721 cells, detected by fluorescent immunocytochemistry and observed by confocal laser scanning microscopy. The positive rate of NET-1 protein expression revealed by immunohistochemistry was 96.9% in HCC（ 126/130）. NET-1 Protein expression in HCC was clearly correlative with HCC cytological variants, there were pronounced higher expressions in clear cell type, pleomorphic cell type, and sarcomatous change than that in hepatocytic type （P 〈 0. 05 ）. NET-1 Protein expression in HCC was positively correlative with the histopathologic grading, clinical stages and HCC with hepatitis and cirrhosis（P 〈0.05） , respectively, and negatively correlated with the presence of patches of necrosis （ P 〈 0.05）. But NET-1 protein expression was not associated with AFP level, tumor size and growth patterns, respectively. Conclusion NET-1 protein is expressed in cytoplasm of HCC cells as irregular granules near Golgi apparatus. NET-1 gene expression may promote the uncontrolled proliferation and abnormal differentiation in HCC cells.