Curcumin effect on proliferation and apoptosis of gastric cancer stem cells via ATK/FoxM1 signaling pathway

doi:10.3969/j.issn.2095-4344.2016.32.003

Abstract

Abstract:

BACKGROUND: Curcumin has crucial inhibitory effects on various cancer cells and cancer stem cells. However, its effect on gastric cancer stem cells and the underlying mechanism of this effect are unclear.
OBJECTIVE: To explore the effect of curcumin on gastric cancer stem cells and the underlying mechanism.
METHODS: Tumor sphere-forming assay and gastric cancer stem cell markers (EpCAM and CD44) were used to separate gastric cancer stem cells from gastric cancer SGC7901 cell lines. Effects of curcumin on the proliferation and apoptosis of gastric cancer stem cells were determined by MTT and flow cytometry analysis, respectively. Western blot analysis was used to detect the expression levels of FoxM1, p-AKT, and AKT. LY294002, an inhibitor of the PI3K/AKT pathway, was used to determine the regulatory relationship between AKT and FoxM1 signaling pathways.
RESULTS AND CONCLUSION: The EpCAM+/CD44+ gastric cancer stem cells were successfully isolated from SGC7901 cells. MTT assay showed that curcumin inhibited the proliferation of gastric cancer stem cells, while flow cytometry analysis showed that curcumin induced apoptosis in gastric cancer stem cells. In addition, the expression levels of p-AKT and FoxM1 were decreased by curcumin treatment. After being treated by LY294002, the expression levels of p-AKT and FoxM1 were down-regulated markedly. In conclusion, curcumin can inhibit cell proliferation and induce apoptosis in gastric cancer stem cells via the ATK/FoxM1 signaling pathway.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Curcumin, Neoplastic Stem Cells, Stomach Neoplasms, Cell Proliferation, Apoptosis, Tissue Engineering

CLC Number:

R394.2

He Dong-li. Curcumin effect on proliferation and apoptosis of gastric cancer stem cells via ATK/FoxM1 signaling pathway[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(32): 4731-4737.

Figures/Tables 2

2.1 胃癌SGC7901干细胞富集结果流式细胞术检测结果显示(图1A)，在胃癌干细胞富集培养前SGC7901细胞中表达EpCAM+/CD44+双阳性细胞的比例为0.9%，富集培养后双阳性细胞的比例为85%。图1B展示了分离到的胃癌干细胞球。 2.2 姜黄素对胃癌干细胞增殖的影响用不同浓度姜黄素(5，10，20 μmol/L)处理胃癌SGC7901干细胞24，48，72 h。MTT实验结果显示，随着药物浓度的增大，其对细胞的抑制作用增强，且随着时间的延长，药物对细胞的抑制作用也呈上升趋势，见图2。可以推测，姜黄素对胃癌干细胞增殖的抑制具有剂量和时间依赖性。 2.3 姜黄素对胃癌干细胞凋亡的影响使用15 μmol/L姜黄素处理胃癌干细胞48 h后收集细胞，流式细胞仪检测细胞的凋亡情况。结果显示，与对照组相比，15 μmol/L姜黄素组胃癌干细胞凋亡数明显增多，差异有显著性意义(图3)。 2.4 姜黄素对胃癌干细胞中FoxM1、p-AKT及AKT蛋白表达的影响胃癌干细胞经15 μmol/L姜黄素处理48 h后，Western blot检测细胞内FoxM1、p-AKT及AKT的蛋白表达情况。结果显示，与对照组相比，姜黄素处理组中p-AKT及FoxM1的蛋白表达水平明显降低(图4)。 2.5 LY294002对胃癌干细胞增殖和凋亡的影响使用AKT信号通路阻断剂LY294002处理胃癌干细胞24 h后，MTT实验检测细胞的增殖和凋亡情况。结果显示，阻断剂处理后，细胞的生存率明显低于对照组(图5A)。细胞凋亡检测结果显示，与对照组相比，阻断剂明显诱导了胃癌干细胞的凋亡(图5B)。2.6 LY294002对FoxM1、p-AKT及AKT蛋白表达的影响 Western blot检测结果显示使用阻断剂处理胃癌干细胞后，与对照组相比，FoxM1及p-AKT的蛋白表达水平明显降低(图6)。可以推测，AKT信号通路的失活降低了FoxM1的蛋白表达水平。"

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