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Archive > Volume 20 Issue 2 > 2013,20(2):153-158. DOI:10.3872/j.issn.1007-385X.2013.02.004 Prev Next
Prompt Report
Virulence factor CagA upregulates TET2 protein expression in human gastric mucosa epithelial GES-1 cells
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Abstract:
Objective: To investigate the association between the expression of the ten-eleven translocation 2 (TET2) protein and the cytotoxin associated gene A (CagA) of Helicobacter pylori (Hp)，and to explore the possible mechanisms of CagA in the process of gastric carcinogenesis. Methods: Real-time PCR was used to detect TET2 mRNA level in human gastric epithelial GES-1 cells and gastric cancer MGC-803 cells. Immuncytochemistry was used to detect the TET2 protein localization and expression in GES-1 and MGC-803 cells. GES-1 cells were transfected with pEGFP-CagA and a cell oxidative stress model was constructed with 200 μmol/L H2O2. Flow cytometry was used to analyze cell cycle and ROS in GES-1 cells. Results: The mean expression level of TET2 mRNA in GES-1 cells was lower than that in MGC-803 cells (1.00±0.08 vs 1.68±0.07, P<0.05). TET2 protein expression in GES-1 cells was lower than that in MGC-803 cells (8.09±3.57 vs 14.60±2.31，P<0.05). Compared with the negative control pEGFP-N1 group, the mean expression level of TET2 mRNA in pEGFP-CagA-transfected GES-1 cells was increased (1.00±0.04 vs 0.06±000，P<005), and TET2 protein expression in pEGFP-CagA-transfected GES-1 cells was also increased (10.82±3.39 vs 16.45±4.40，P<0.05), and a higher level of ROS production was observed in pEGFP-CagA-transfected GES-1 cells (69±90 vs 91±16.8，P<0.05), which significantly displayed the cell apoptosis in cycle analysis. In the cell oxidative stress model, TET2 mRNA level in H2O2 treated cells was higher than that in normal GES-1 cells (1.44±0.02 vs 1.00±0.04，P<005) and TET2 protein expression level was higher than that in normal GES-1 cells (11.74±4.34 vs 15.72±452，P<0.05). Conclusion: CagA factor can induce ROS accumulation and cell cycle disruption of GES-1, indicating that TET2 can be upregulated by oxidative stress and may be involved in the progress of CagA-induced carcinogenesis.
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Project Supported:
Project supported by the National Key Basic Research and Development Program (973 Program) of China （No. 2010CB933901), and the National Science Fundation for Distinguished Young Scholar of China （No. 81225010）
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