Wang Q, Shen W, Tao G-Q, Sun J, Shi L-P
Department of General Surgery, Wuxi People's Hospital, Jiangsu Province, China.
Eur Rev Med Pharmacol Sci. 2017 Mar;21(5):1006-1012.
This study is to investigate the effect of low concentration hydrogen peroxide (H2O2) on the proliferation of gastric cancer AGS cell line in vitro and the mechanism.
AGS cells were treated with different low concentrations of H2O2 (1, 0.1, 0.01, and 0.001 μm) for 48 hours. The effect of H2O2 concentration gradient on the activity of AGS cell activities was detected by methyl thiazolyl tetrazolium (MTT) method. The expression of the epidermal growth factor receptor (EGFR) and its downstream signaling pathway extracellular signal-regulated kinase (ERK) protein in H2O2 was detected by Western blot method; moreover, the effect of H2O2 on intracellular reactive oxygen species (ROS) in AGS cells was observed under the fluorescence microscope and quantitative analysis by flow cytometry. The effect of H2O2 on the level of c-myc mRNA in AGS cells was also detected by reverse transcription polymerase chain reaction (RT-PCR).
MTT detection results showed that 1 μm and 0.1 μm H2O2 at 48h can effectively promote the proliferation of AGS cells (p<0.05). The Western blot results showed that after 48 hours of low concentration of H2O2 treatment of AGS cells, the EGFR protein levels and ERK protein phosphorylation levels increased significantly (p<0.05). Fluorescence microscopy and flow cytometry showed that the low concentration of H2O2 increased the intracellular reactive oxygen species (ROS). RT-PCR results showed the levels of c-myc mRNA in AGS cells treated with a low concentration of H2O2 were significantly increased (p<0.05) and EGFR blocking agent AG1478 can resist the above effects.
Low concentration of H2O2 can significantly promote the proliferation of AGS cells by activating EGFR/ERK signaling pathway.
本研究旨在探讨低浓度过氧化氢（H2O2）对胃癌AGS细胞系体外增殖的影响及其机制。
将AGS细胞用不同低浓度的H2O2（1、0.1、0.01和0.001μm）处理48小时。采用甲基噻唑基四氮唑（MTT）法检测H2O2浓度梯度对AGS细胞活性的影响。用蛋白质免疫印迹法检测H2O2中表皮生长因子受体（EGFR）及其下游信号通路细胞外信号调节激酶（ERK）蛋白的表达；此外，在荧光显微镜下观察H2O2对AGS细胞内活性氧（ROS）的影响，并通过流式细胞术进行定量分析。还用逆转录聚合酶链反应（RT-PCR）检测H2O2对AGS细胞中c-myc mRNA水平的影响。
MTT检测结果显示，48小时时1μm和0.1μm的H2O2能有效促进AGS细胞的增殖（p<0.05）。蛋白质免疫印迹结果显示，低浓度H2O2处理AGS细胞48小时后，EGFR蛋白水平和ERK蛋白磷酸化水平显著升高（p<0.05）。荧光显微镜和流式细胞术显示，低浓度H2O2增加了细胞内活性氧（ROS）。RT-PCR结果显示，低浓度H2O2处理的AGS细胞中c-myc mRNA水平显著升高（p<0.05），且EGFR阻断剂AG1478可抵抗上述作用。
低浓度H2O2可通过激活EGFR/ERK信号通路显著促进AGS细胞增殖。
