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目的:研究三氧化二砷(As2O3)对人胃腺癌细胞株SGC-7901的生物效应及其对线粒体和半胱氨酸蛋白酶家族-3(caspase-3)的作用。方法:通过MTT比色实验检测不同浓度As2O3对该细胞株的生长抑制作用;经Hoechst 33258染色后用荧光显微镜观察细胞核的形态变化;经过细胞线粒体膜电位检测区分凋亡细胞与正常细胞,并经流式细胞仪分析;caspase-3吸光度检测法测定As2O3组caspase-3的活化程度。结果:As2O3明显抑制SGC-7901人胃腺癌细胞的生长,抑制作用的强度呈时间依赖性(方差分析,P<0.01);Hoechst 33258染色后荧光显微镜观察细胞核固缩碎裂边集呈强蓝色荧光;线粒体膜电位检测法,流式细胞仪检测法,caspase-3吸光度检测法均检测到胃腺癌细胞的凋亡。结论:As2O3破坏线粒体跨膜电位和激活caspase-3活性可能是As2O3诱导人胃腺癌SGC-7901细胞凋亡的重要机制。
Objective: To investigate the biological effect of arsenic trioxide (As2O3) on human gastric adenocarcinoma cell line SGC-7901 and its effect on mitochondria and caspase-3. Methods: MTT colorimetric assay was used to detect the growth inhibition of As2O3 at different concentrations. The morphological changes of nuclei were observed by fluorescence microscopy after Hoechst 33258 staining. Apoptotic cells and normal cells were differentiated by mitochondrial membrane potential Flow cytometry analysis; caspase-3 absorbance assay Assay caspase-3 activation level in As2O3 group. Results: As2O3 significantly inhibited the growth of SGC-7901 human gastric adenocarcinoma cells in a time-dependent manner (ANOVA, P <0.01). The fluorescence intensity of Hoechst 33258 staining showed a strong blue Fluorescence; Apoptosis of gastric adenocarcinoma cells was detected by mitochondrial membrane potential assay, flow cytometry and caspase-3 absorbance assay. CONCLUSION: As2O3 can damage mitochondrial transmembrane potential and activate caspase-3 activity, which may be an important mechanism of As2O3-induced apoptosis in human gastric adenocarcinoma SGC-7901 cells.