Differential Proteomics Research In Heterogeneity Of Differentiation Of Hepatocellular Carcinoma

Objective: Manifestation of tumor heterogeneity is complexly and diversely. It shows a huge difference even for the same type of tumor, among different individuals or the same individual in different pathological period, especially the difference in structure of tumor. The heterogeneity of tumor has brought tremendous challenges for tumor’s early diagnosis, treatment and prognosis. From the perspective of heterogeneity to recognize and study the tumor’s intrinsic mechanism. To study the molecular mechanisms of differentiation of hepatocellular carcinoma by using proteomics and bioinformatics methods. It will provide experimental and theoretical evidence to confirm tumor biomarkers about the differentiation of hepatocellular carcinoma associated, understanding of liver cancer and to enhance the diagnosis and treatment of patients with liver cancer.Materials and Methods: Fresh hepatocellular carcinoma tissues(HBsAg+) were collected on hepatectomy from The First Hospital of Lanzhou University. The degree of cellular differentiation was classified into groups according to the postoperative pathological diagnosis: moderately/poorly-differentiated group and well-differentiated group. Protein samples were extracted from tissues and total protein amount in each sample was assayed. Proteins were separated by proteomics technologies(two-dimensional gel electrophoresis, 2DE), and identified by MALDI-TOF-MS. Find out the expressed proteins which associated the heterogeneity of hepatocellular carcinoma. Through systems biology and bioinformatics pathway speculates the functions in different proteins. Using immunohistochemical method to verify the results compared with proteomics methods and clarify different expression of proteins which associated in differentiation of liver cancer.Result: Eleven proteins were identified successfully, in which six of them were overexpressed in moderately/poorly-differentiated tumor tissues group: Peroxiredoxin-2( PRDX2), Heat shock protein beta-1(HSPB1), Haptoglobin(HP), AnnexinA2(ANXA2), Enoyl-CoA hydratase mitochondrial(ECHM), Glutathione S-transferase Omega-1(GSTO1); and five of them were correspondingly down-regulated: Calreticulin(CALR), Fructose-bisphosphate aldolase a(ALDOA), Fibrinogen beta chain(FIBB), Serotransferrin(TRFE), Hemoglobin subunit beta(HBB). After analysis by biological information, those differential proteins involved in: regulation of heat shock protein, control of multiple signaling pathways, glycolysis of glucose metabolism, etc. Four kind of proteins were chosen for immunohistochemical verification. The results were consistent with proteomics research, PRDX2, HSPB1, ANXA2 were overexpressed in poorly-differentiated HCC tissues. CALR was down-regulated in poorly-differentiated HCC tissues.Conclusion: Confirmed that ANXA2, PRDX2 and HSPB1 were overexpressed in poorly-differentiated hepatocellular carcinoma tissue prompts high invasive characteristics. May serves as biomarkers for early detection of liver cancer metastasis. CALR extreme expression in well-differentiated hepatocellular carcinoma which may be associated with CALR immune mediated phagocytosis, and could be a potential target for gene therapy of liver cancer.