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The Mechanism of Helicobacter Pylori Promote the HepG2 Cell Proliferation

Author: ChenZhiTian
Tutor: ChenYanLing
School: Fujian Medical
Course: Surgery
Keywords: hepatocellular RT-PCR helicobacer GENE CHIPS
CLC: R735.7
Type: Master's thesis
Year: 2008
Downloads: 54
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Abstract

Objectives:To testify whether the Helicobacter pylori could promote the cell proliferation of HepG2 cells and then discover the mechanisms in order kown the progress of Helicobacter pylori participate the hepatoeellular carcinoma incur.Method:1. HepG2 cells and H.pylori Was co-cultured in several concentration and compound the Cell Counting Kit-8 agent with it after 24 hours to Measure the absorbance at 450 nm using a microplate reader to inspect the change of HepG2 cells proliferation and discover the best dose。2. inspect the change of the gene that pick out from the expression in HepG2 cells by the Affymetrix GENE CHIPS.3.Add the H.pylori in the concentration that could promote the cells proliferation of HepG2 cells and co-cultured it and 24h.the total RNA of untreated and H.pylori treated HepG2 cells RNA were separated and RT-PCR was performed to detect that the change of genes that pick out from GENE CHIPS expression to confirm the result of it.Result: 1. When HepG2 cells and H.pylori Was co-cultured in the MOI of (0.3:1～0.075:1), the the HepG2 cells proliferation was great promoted by H.pylori.2.The GENE CHIPS shows that there are 19 genes up-regulation and 16 genes down-regulation after the H.pylori was compound in the HepG2 cells. They were contain 6 cytoskeleton up genes that intercellular adhesion molecule 1(ICAM-1)、transgelin、talin 1、tubulin、Integrin及keratin 23。Also they contain 8 DNA binding protein genes that tubulin beta polypeptide paralog、Cell division cycle 2-like 5、myelin expression factor 2、V-erb-a erythroblastic leukemia viral oncogene homolog 4 were up regulation,and 4 down regulation gene they are zinc finger protein 513、nuclear receptor subfamily 1(group I, member 3)、Clock homolog、SRY (sex determining region Y)-box 5。At last 2 cytokine up regulation genes were interleukin 8 and interleukin 1 family, member 9。3.The RT-PCR result reveal that there are many difference between the five differential GENE expression were successfully identified, These genes included Tubulin、ICAM、ARTS、Akeratin、ARHGDIA.Conclusions:1.The HepG2 cells proliferation was great enhance after compound with the H.pylori in a suitable MOI。2.The GENE CHIPS shows that there are many genetic expression change after HepG2 cells was compound with the H.pylori in an suitable MOI. They may include 6 cytoskeleton up genes,8 DNA binding protein genes ,2 cytokine up regulation genes and they may inclosed the progress to the hepatoeellular carcinoma incur。3.There is a significant difference at the genes expression when the hepG2 cells and H.pylori Was co-cultured ,this genes may be include the progress of H.pylori promote the HepG2 cells proliferation and even enforce the progress of the carcinogenesis of hepatoeellular carcinoma.

The Mechanism of Helicobacter Pylori Promote the HepG2 Cell Proliferation

Abstract

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