幽门螺杆菌促肝细胞增殖及分子机制的初步研究

Effects of H.pylori on proliferation of hepatocytes and the molecular mechanism

目的研究幽门螺杆菌(Hp)对肝细胞增殖的影响并探讨其分子机制。方法以不同量的Hp标准株NCTC11637与HepG2肝细胞共培养,CCK-8法检测细胞的增殖情况并确定最佳Hp作用剂量。用Affymetrix人类基因表达谱芯片检测Hp与HepG2共培养对HepG2细胞基因表达谱的影响,并选取5个差异表达基因以半定量RT-PCR技术验证。结果 MOI比值(细菌与细胞个数比)在0.15∶1～0.075∶1时,Hp对HepG2细胞具有明显的促增殖作用;HepG2细胞经MOI比值为0.15∶1Hp处理后有35个基因转录发生变化:ICAM-1等19个基因上调,SLC38A4等16个基因下调,这些变化基因涉及细胞骨架/基质、DNA结合蛋白及转录因子等;5个基因表达情况与表达谱芯片检测结果完全吻合。结论一定剂量的幽门螺杆菌可促进肝细胞增殖,这与Hp导致HepG2细胞基因转录调控有关基因的表达变化有关。

Objective To study the effects of H. pylori （Hp） on proliferation of hepatocytes and explore the molecular mechanism. Methods Hp strain NCTC11637 was co-cultured with HepG2 hepatoblastoma cells. The proliferation of cells was evaluated by CCK-8 assay and to determine Hp best dose. The effects of Hp on the gene expression profile of HepG2 cells were analyzed by using Affymetrix human oligonucleotide genechip. The chip results were validated by semi-quantitative RT-PCR using randomly selected 5 genes. Results Hp enhance the proliferation of HepG2 cells with the MOI ranging from 0.15：1 to 0. 075：1 ,the gene chip analysis showed that totally 35 genes were changed after the treatment of HepG2 cells with Hp, 19 of them including ICAM1 （intercellular adhesion molecule 1） were up-regula- ted, and 16 of them including SLC38A4 （solute carrier family 38, member 4） were down-regulated. The majority of these changed genes could be categorized to the classes of cellular skeleton/matrix, DNA binding proteins and ranscriptional factors. In addition, the semi-RT-PCR results fit that of gene chip analysis. Conclusions Hp could enhance the proliferation of HepG2 ceils, and this effects are associated with the changes of transcription regulation related genes,