Yu Yao, Yang Yu, Wang Jing
Department of Oncology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China.
Oncol Rep. 2014 Sep;32(3):973-8. doi: 10.3892/or.2014.3276. Epub 2014 Jun 20.
The present study aimed to investigate the effect of arsenic trioxide (As2O3) on human gastric cancer SGC-7901 cells. SGC-7901 cells were treated with different concentrations of As2O3 in the cell growth media for 24, 48 and 72 h, and the growth rates were determined by WST-1 cell proliferation assays. Analyses of nuclear morphological changes were performed with DAPI fluorescence staining. Cell apoptosis rates as assessed by flow cytometry were determined after cells were grown in media for 48 h containing different As2O3 concentrations. The protein expression patterns of the apoptosis factors, Bax, Fas and caspase-8, and anti-apoptosis factors, Akt, p-Akt, mTOR and p-mTOR, were evaluated by western blot analysis following treatment of the cells with different As2O3 concentrations in the cell growth media for 48 h. As a result, As2O3 inhibited the growth of human gastric cancer SGC-7901 cells in concentrations >5 µmol/l for longer than 24 h. Flow cytometric analysis revealed that the apoptosis of SGC-7901 cells occurred in an As2O3 concentration-dependent manner after 48 h (P<0.001). Expression levels of Bax, Fas and caspase-8 were increased, whereas expression levels of Akt, p-Akt, mTOR and p-mTOR were decreased in the SGC-7901 cells after a 48-h incubation with different As2O3 concentrations. In conclusion, As2O3 induced human gastric cancer SGC-7901 cell apoptosis in a time- and concentration dependent manner by inhibiting the activity of anti-apoptosis-related factors.
本研究旨在探讨三氧化二砷（As2O3）对人胃癌SGC-7901细胞的影响。在细胞生长培养基中用不同浓度的As2O3处理SGC-7901细胞24、48和72小时，并通过WST-1细胞增殖试验测定生长速率。用DAPI荧光染色进行核形态变化分析。在含有不同As2O3浓度的培养基中培养细胞48小时后，通过流式细胞术测定细胞凋亡率。在用不同As2O3浓度的细胞生长培养基处理细胞48小时后，通过蛋白质印迹分析评估凋亡因子Bax、Fas和caspase-8以及抗凋亡因子Akt、p-Akt、mTOR和p-mTOR的蛋白质表达模式。结果，As2O3在浓度>5µmol/l且处理时间超过24小时时抑制人胃癌SGC-7901细胞的生长。流式细胞术分析显示，48小时后SGC-7901细胞的凋亡呈As2O3浓度依赖性（P<0.001）。在用不同As2O3浓度孵育48小时后，SGC-7901细胞中Bax、Fas和caspase-8的表达水平升高，而Akt、p-Akt、mTOR和p-mTOR的表达水平降低。总之，As2O3通过抑制抗凋亡相关因子的活性，以时间和浓度依赖性方式诱导人胃癌SGC-7901细胞凋亡。
