目的 探讨裙带菜多糖硫酸酯S-UPPSⅠB体外诱导人肝癌HepG-2细胞凋亡的作用及机制。方法 采用四甲基偶氮唑蓝(MTT)法检测S-UPPSⅠB对HepG-2细胞增殖作用的影响;激光共聚焦显微镜测定HepG-2细胞内[Ca2+]i水平;流式细胞仪测定细胞凋亡率及Bcl-2、Bax、Cyt-c、p53的表达;Caspase-3, -9试剂盒检测蛋白表达。结果 S-UPPSⅠB 抑制HepG-2细胞的IC50值为50.09 μgmL-1, 随着给药剂量的增加, 凋亡率亦增加, 并且对Ca2+及其通路的相关蛋白有明显的调节作用, 其中[Ca2+]i的水平及Cyt-C、Caspase-3, -9、p53的表达均有显著增加(P<0.05), Bcl-2/Bax比值降低。结论 裙带菜多糖硫酸酯S-UPPSⅠB可显著抑制人肝癌HepG-2细胞增殖, 并可通过启动线粒体途径诱导HepG-2细胞发生凋亡。
Abstract
OBJECTIVE To investigate the mechanism of human liver cancer HepG-2 cells apoptosis induced by Undaria pinnatifida sulfated polysaccharides S-UPPSⅠB. METHODS The anti-proliferation effects of S-UPPSⅠB on HepG-2 cells was by MTT assay. [Ca2+]i in HepG-2 cells was detected by laser cofocal scaning microscopy (LCSM). The apoptosis rate and protein expression level of Bcl-2, Bax, Cyt-C and p53 were detected by flow cytometry (FCM). Caspase Assay Kit was used to detected the activities of Caspase-3 and -9. RESULTS S-UPPSⅠB could inhibit the proliferation of HepG-2 cells and the IC50 was 50.09 μgmL-1. With the increase of drug delivery dosage, apoptosis rate also increased, and the significant regulating effects of S-UPPSⅠB on Ca2+ and its associated channel proteins were observed. The [Ca2+]i level, the protein expression level of Cyt-c, p53 and the activities of Caspase-3 and -9 were all increased remarkably (P<0.05). The ratio of Bcl-2 to Bax was reduced. CONCLUSION Undaria pinnatifida sulfated polysaccharides S-UPPSⅠB can effectively inhibit the proliferation of human liver cancer HepG-2 cells by inducing apoptosis of HepG-2 cells through mitochondrial pathway.
关键词
裙带菜 /
多糖硫酸酯 /
人肝癌HepG-2细胞 /
凋亡
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Key words
Undaria pinnitafida /
sulfated polysaccharide /
HepG-2 /
apoptosis
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中图分类号:
R965
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参考文献
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脚注
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基金
黑龙江省教育厅面上项目资助(12521134);国家级大学生创新训练计划项目资助(201610240025)
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