Chen Xiao-Li, Zhou Le, Yang Jun, Shen Fu-Kun, Zhao Shi-Ping, Wang Yi-Li
Department of Pathology, the Second Affiliated Hospital, School of Medicine, Xian Jiaotong University, Xian, 710004 Shaanxi, P.R. China.
Mol Med Rep. 2010 Jul-Aug;3(4):589-96. doi: 10.3892/mmr_00000302.
Hepatocellular carcinoma (HCC) is one of the most common types of cancer worldwide. The initial hepatocellular alterations that precede the appearence of HCC include chronic viral hepatitis/cirrhosis, foci of phenotypically altered hepatocytes and, subsequently, dysplastic hepatocytes that form foci and nodules. These changes cause a discrepancy in the microenvironment of liver cells, which may result in changes in the protein expression profile of the cells. The aim of the present study was to investigate differences between the protein expression profiles at various stages of liver disease in order to better understand the mechanisms of HCC and to identify potential biomarkers for its early diagnosis. The proteins of specific cells were obtained from HCC tissue sections and pre-cancerous lesions using a manual microdissection technique, and were investigated by a two dimensional gel electrophoresis (2-DE) MALDI-TOF MS proteomics approach. Select identified proteins were reconfirmed by immunohistochemistry. A total of 95 differentially expressed proteins, with an over 2-fold disparity in expression levels between cells of varying morphology during the stages of hepatocarcinogenesis, were detected by 2-DE. Among these 95 proteins, 80 were determined to be involved in numerous cell functions, including cell growth and proliferation, protein synthesis and metabolism, apoptosis and signal transduction. These identified proteins, which include stratifin (14-3-3), transgelin 2, heat-shock protein (HSP)70, HSP27, manganese superoxide dismutase, prohibitin, DJ1, α-enolase, peroxiredoxin 6, aldo-keto reductase family member B10, phosphoglycerate kinase 1, α-1-antitrypsin and nm23-H1, may play a role in the development of HCC. Protein expression profiles differed markedly between the HCC tissue samples and pre-cancerous lesions, suggesting that alterations in protein expression occurred frequently during the process of hepatocarcinogenesis. Analysis of the differential expression of proteins related to the development of HCC may help elucidate the molecular mechanisms of the disease. These proteins may also serve as candidate biomarkers for early HCC diagnosis.
肝细胞癌（HCC）是全球最常见的癌症类型之一。在HCC出现之前的初始肝细胞改变包括慢性病毒性肝炎/肝硬化、表型改变的肝细胞灶，随后是形成灶和结节的发育异常肝细胞。这些变化导致肝细胞微环境出现差异，这可能会导致细胞蛋白质表达谱发生变化。本研究的目的是调查肝病各个阶段蛋白质表达谱之间的差异，以便更好地了解HCC的发病机制，并识别其早期诊断的潜在生物标志物。使用手动显微切割技术从HCC组织切片和癌前病变中获取特定细胞的蛋白质，并通过二维凝胶电泳（2-DE）基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）蛋白质组学方法进行研究。通过免疫组织化学对选定的已鉴定蛋白质进行重新确认。二维凝胶电泳检测到总共95种差异表达蛋白，在肝癌发生阶段，不同形态的细胞之间表达水平存在超过2倍的差异。在这95种蛋白质中，有80种被确定参与多种细胞功能，包括细胞生长和增殖、蛋白质合成和代谢、细胞凋亡和信号转导。这些已鉴定的蛋白质，包括层粘连蛋白（14-3-3）、原肌球蛋白2、热休克蛋白（HSP）70、HSP27、锰超氧化物歧化酶、抑制素、DJ1、α-烯醇化酶、过氧化物还原酶6、醛酮还原酶家族成员B10、磷酸甘油酸激酶1、α-1抗胰蛋白酶和nm23-H1，可能在HCC的发展中起作用。HCC组织样本和癌前病变之间的蛋白质表达谱存在明显差异，这表明在肝癌发生过程中蛋白质表达的改变频繁发生。分析与HCC发展相关蛋白质的差异表达可能有助于阐明该疾病的分子机制。这些蛋白质也可能作为HCC早期诊断的候选生物标志物。
