【摘要】 目的探讨藤黄酸(GA)颗粒对结肠癌干细胞增殖和凋亡的影响。方法通过体外培养结肠癌HCT116细胞,采用免疫磁珠标记分选技术从HCT116细胞中分选出ALDH1~+/CD133~+结肠癌干细胞;经不同浓度(0、0.5、1和2 mg/ml)GA颗粒处理结肠癌干细胞后,采用克隆形成实验观察细胞的克隆能力,CCK-8法检测细胞的增殖能力,流式细胞仪检测细胞的凋亡情况,Western blotting及实时定量PCR(QPCR)检测干细胞中Sox2和Oct4蛋白及m RNA的表达。结果免疫磁珠标记分选技术成功分选出符合要求的ALDH1~+/CD133~+结肠癌干细胞;克隆形成实验结果显示,不同浓度GA颗粒处理后细胞克隆形成率显著降低,且呈剂量依赖性;CCK-8检测发现,GA颗粒能够呈剂量-时间依赖性地抑制结肠癌干细胞的增殖;流式细胞仪检测结果表明,GA颗粒能够呈剂量依赖性地诱导结肠癌干细胞的凋亡;Western blotting及QPCR检测结果显示,GA颗粒能够呈剂量依赖性地降低Oct4和Sox2蛋白及m RNA水平。结论 GA颗粒能够明显抑制结肠癌干细胞的克隆、增殖及Oct4和Sox2的表达并诱导细胞凋亡。更多还原

【Abstract】 Objective To investigate the effect of gambogic acid( GA) granules on proliferation and apoptosis of colon cancer stem cells. Methods Colon cancer HCT116 cells were cultured in vitro and ALDH1~+/CD133~+colon cancer stem cells were isolated from HCT116 cells by immunomagnetic beads sorting technique. Exposure to different concentrations( 0,0. 5,1,2 mg/ml) of GA granules,the colony formation ability of colon cancer stem cells was detected by colony forming assay,and cell proliferative ability was detected by CCK-8 assay. The apoptosis of the cells was detected by flow cytometry,and Western blotting and QPCR were used to detect the protein and m RNA levels of Sox2 and Oct4 in stem cells. Results ALDH1~+/CD133~+colon cancer stem cells were successfully isolated by immunomagnetic beads sorting technique. Cloning formation assay revealed that cell clone formation rate was decreased significantly in a dose-dependent manner after treatment with different concentrations of GA granules. CCK-8 detection showed that GA granules could inhibit the proliferation of colon cancer stem cells in a time-and dose-dependent manner. Flow cytometry results showed that GA granules could induce apoptosis of colon cancer stem cells in a dose-dependent manner. Western blotting and QPCR results showed that GA granules could decrease the protein and m RNA levels of Oct4 and Sox2 in stem cells in a dose-dependent manner. Conclusion GA granules can inhibit colon cancer stem cell proliferation,cloning,expression of Sox2 and Oct4,and induce cell apoptosis.更多还原