Expression And Preliminary Functional Analyses Of TRIM3 In Primary Hepartocellular Carcinoma

Background and PurposesHepatocellular carcinoma(HCC) is a common malignant tumor, ranked as the fifth in global malignant tumor incidence. HCC is the fourth causing of the malignant tumor related death in many countries. Even with aggressive treatments, HCC still has a poor prognosis, with a 5-year survival rate as low as 25%-39%. The leading causes of HCC with poor prognosis are high malignant degree, vasal aggression and liver transfer. The emergence of HCC is related with many factors, the genetic factors is the most major in overexpression of oncogene and tumor-suppressor gene passivation. For purpose of improving the outcome of patients with liver cancer, it is essential to find additional molecular points of early diagnosis and effective therapy for liver cancer.TRIM3 protein, either called BERP protein, was originally as alpha auxiliary actin binding protein 4 be cloned, but the specific function of it is not very clear at present. The name of the TRIM is because of the distinctive features of the TRIM family with three domain structure. The three structure domain from the N end to C was followed by zinc finger structure, 1 or 2 B- box, a coiled coil structure domain. In recent years, there are several studies have confirmed that the TRIM family members of the protein is hardly correlated with the development of tumor, the research has confirmed TRIM3 in malignant glioma is a new tumor suppressor genes, act a part of the tumor restrain gene by inhibiting p21 protein. Lower TRIM3 expression can anvance p21 gene expression in cells increased, and can promote the progress of tumor cell proliferation and transplantation and transplantation of the becoing of the tumor blood vessels. Overexpression of TRIM3 can make the cells in the cell cycle phase G0/1 accumulation, occurring to cell growth arrest. So think TRIM3 may be a potential tumor suppressor genes. However, the role of the TRIM3 in liver cancer still ill-defined.The purpose of this topic is find the correlation with clinical parameters and prognosis of the tumor patients through the detection of TRIM3 gene expression in liver cancer, and preliminary study TRIM3 biological function on liver cancer.MethodsReal-time quantitative RT-PCR, western blotting and immunohistochemistry technique to detect liver cancer cell lines and TRIM3 expression in liver cancer tissue samples.In hepatocellular carcinoma cell line transfection TRIM3 gene and protein expression TRIM3, discuss the cell growth(proliferation and clone formation), cell cycle, apoptosis, cell transfer(migration and invasion) aspects of the biological functions.Statistical methods:Paired t test is mainly used for liver cancer tissue and normal tissue adjacent to carcinoma TRIM3 transcription and protein expression differences; Chi-square test is mainly used for high expression and low expression group differences between clinical parameters; Survival analysis is used to compare high expression and low expression group survival time; Cox regression analysis was used to study the related factors affecting survival. Results1. In paired samples, the expression of TRIM3(P = 0.018) m RNA transcription level and protein level(P = 0.02) in cancerous tissue was remarkablely decrease than adjacent normal tissue.Compared with LO2 cell, TRIM3 m RNA and protein expression in liver cancer cell have different degrees of cut, especially in Hep G2 and Bel7402 cell strains.2. Immune histochemical methods examined 129 cases of paraffin embedding TRIM3 protein expression in the organization, TRIM3 is mainly lied on cytoplasm, relative to the lower utterance of cancerous samples, adjacent normal tissue for high expression;TRIM3 expression and tumor size(P = 0.034), pathological grading(P < 0.001), TNM stage(P = 0.021) and serum AFP level(P = 0.025);Survival analysis revealed that TRIM3 lower utterance suggests malignant prognosis;TRIM3 lower utterance may independently be an adverse prognostic factors for liver cancer patients.3.Build lentivirus vector and transfected Hep G2 and Bel7402 cells, found TRIM3 inhibits cell proliferation, reduce the clone formation, migration and invasion ability, causes the G1 phase cell arrest, inducing tumor cell apoptosis. ConclusionTRIM3 acts as crucial role on the appearance of liver cancer. It can be used as the points for the targeted therapy of liver cancer, also can be accessed as an isolated prognostic factor of liver cancer.