Dissertation > Excellent graduate degree dissertation topics show

Effect of HBx Gene on Endogenous Survivin Expression in Human Hepatoma Cells

Author: LiJinLing
Tutor: HeYan
School: Central South University
Course: Internal Medicine
Keywords: Survivin HBVx gene HCC
CLC: R735.7
Type: Master's thesis
Year: 2008
Downloads: 78
Quote: 0
Read: Download Dissertation

Abstract

Purpose of survivin (survivin) is a member of the inhibitor of apoptosis protein family (IAP), X gene is the the HBV genome smallest open reading frame (ORF) of HCC development is most closely related. To explore X gene endogenous survivin expression as well as analysis and comparison of survivin expression in hepatitis, cirrhosis of the liver and tumor tissues, and further explore the survivin into possible molecular mechanisms of HCC development in patients with chronic hepatitis B infection by HBV laid theoretical and experimental foundation. Research methods with liposomes Lipofectmine ^{TM 2000 respectively pcDNA3.1-HBx (the expression X gene) and empty the plasmid pcDNA3.1 import HepG2 cells, and the other to set up a group of untransfected control of any plasmid blank . G418 were given 48 hours after transfection screening to establish stable cell lines, the following experiment: (1) application of immunocytochemical techniques to detect HBx expression. (2) Application of immune cells Chemical Technology qualitative detection of the experimental group and the control group survivin expression. (3) extraction of the cells in each group total protein comparison groups survivin expression by Western blot (Western Bloting) method, and further confirms the results of the cellular immune technology. (4) our hospital 20 patients with pathologically confirmed hepatocellular carcinoma tissue specimens (nine cases in which poorly differentiated, moderately differentiated seven cases, four cases of well-differentiated), and 30 cases of hepatitis tissue samples (including early cirrhosis of the liver tissue 10 cases, pathological grade: G2/S2 G4/S4), detection of survivin expression by immunohistochemical techniques. Immunocytochemical method results transfected with plasmid pcDNA3.1-HBx cells HBx expression, brown particles distributed in the cytoplasm. Transfected pcDNA3.1 group and untransfected group showed no expression prove plasmid transfection success. The transfected cells were screened by G418 resistant clones 20 days (final concentration maintaining of 400μg ╱ ml), namely the establishment of a stable cell lines. The immunocytochemical method shows three groups of cells have Survivin expression the brown particles distributed in the cytoplasm, but pcDNA3.1-HBx plasmid is transfected cells Survivin protein expression is the most obvious, compared with transfection of pcDNA3.1 group and the control group was statistically significant (P <0.05) significantly. Wstern Bloting detection Survivin protein expression levels of cells transfected with the X gene expression plasmid pcDNA3.1-HBx cells Survivin expression was significantly higher than that transfected with pcDNA3.1 group and untransfected group, further confirms the results of immunocytochemistry . X gene can promote endogenous Survivin expression. Immunohistochemical method to 20 cases of liver cancer tissues, 17 cases of Survivin protein expression (17/20, the positive rate was 85%), brown particles distributed in the cytoplasm of hepatoma cells, positive cells in HCC were scattered in the distribution , of which 58.8% (10/17) were strongly positive expression, 23.5% (4/17) showed a positive expression, 17.7% (3/17) weakly positive expression, the expression of positive intensity considering their corresponding pathological degree of malignancy staging. Two cases of 20 cases of hepatitis organization Survivin protein expression (2/20, the positive rate was 10%) and three cases of 10 patients with early cirrhosis organization Survivin protein expression (3/10, the positive rate was 30%). The HCC tissues Survivin expression level significantly higher than hepatitis organizations and early cirrhosis organization, the differences were statistically significant (P <0.01). Conclusion containing X gene expression pcDNA3.1-HBx plasmid transfected into HepG2 cells, and the establishment of a stable cell line; X gene can promote endogenous survivin protein expression, which may HBV infection, chronic hepatitis B patients developed HCC one of the molecular mechanisms; Survivin protein in different tissue specificity as the HCC diagnostic indicators and a new target for drug research governance.}

Effect of HBx Gene on Endogenous Survivin Expression in Human Hepatoma Cells

Abstract

Related Dissertations