Quercetin inhibits proliferation and promotes apoptosis of glioma stem cells via the STAT pathway

doi:10.3969/j.issn.2095-4344.2017.05.001

Abstract

Abstract:

BACKGROUND: Several studies have reported that quercetin can inhibit the proliferation and migration but promotes apoptosis of tumor cells.
OBJECTIVE: To explore the effect of quercetin on the proliferation and apoptosis of glioma stem cells and the signal pathway involved.
METHODS: Glioma stem cells were isolated by immunomagnetic beads and treated in culture medium containing different concentrations of quercetin (0, 25, 50, 100 μmol/L). Cell proliferation was measured by MTT and apoptosis measured by flow cytometry at 48 hours after culture. The expression levels of Bcl-2, Bax and survivin, which are related to apoptosis, were detected by western blot. The expression of proliferating cell nuclear antigen (PCNA), which is related to proliferation, was also detected by western blot. The expression of STAT3 and p-STAT3 was also determined by western blot.
RESULTS AND CONCLUSION: (1) Compared with the control (0 μmol/L) group, quercetin inhibited the proliferation of glioma stem cells in a dose-depended manner (P < 0.05). With the increase of the concentration of quercetin, the expression of PCNA was increased (P < 0.05). (2) Quercetin induced apoptosis of glioma stem cells dose-dependently (P < 0.05). With the increase of the concentration of quercetin, the expression of Bcl-2 and survivin was decreased, while the expression Bax was increased. (3) Quercetin could also inhibit phosphorylation of STAT3 dose-dependently, but the level of STAT3 was not changed. To conclude, these results show that quercetin could inhibit the proliferation of glioma stem cells and promote apoptosis via the STAT 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: Drugs, Chinese Herbal, Neoplastic Stem Cells, Cell Proliferation, Apoptosis

CLC Number:

R394.2

Liu Yi-feng, Zhang Bao-chao, Wen Chang-ming, Wen Gong-ling, Zhou Gou-ping, Zhang Jing-wei, He Hai-fa, Wang Ning, Li Wei. Quercetin inhibits proliferation and promotes apoptosis of glioma stem cells via the STAT pathway[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(5): 657-662.

Figures/Tables 1

2.1 槲皮素抑制胶质瘤干细胞的增殖与对照组比较，槲皮素处理胶质瘤干细胞后，胶质瘤干细胞的增殖受到抑制，并且槲皮素对胶质瘤干细胞的增殖抑制作用呈剂量依赖性(P < 0.05)，见图1。增殖细胞核抗原与细胞的DNA合成密切相关，是良好反映细胞增殖的标志物。研究中发现不同浓度槲皮素处理胶质瘤干细胞后，增殖细胞核抗原表达随槲皮素浓度的升高而降低，见图1，表明槲皮素抑制了胶质瘤干细胞的增殖。 2.2 槲皮素诱导胶质瘤干细胞的凋亡流式细胞仪检测结果显示，与对照组比较，槲皮素处理胶质瘤干细胞后，胶质瘤干细胞中凋亡细胞数增加，并且随着槲皮素浓度的增加凋亡细胞数逐渐增加(P < 0.05)，见图2。Western blot检测结果显示，槲皮素处理胶质瘤干细胞后，随着槲皮素浓度的增加，凋亡促进蛋白Bax表达逐渐升高，凋亡抑制蛋白Bcl-2和存活素表达逐渐降低，见图2。这些结果表明槲皮素可以促进胶质瘤干细胞的凋亡。 2.3 槲皮素抑制胶质瘤干细胞p-STAT3的表达与对照组比较，槲皮素处理后胶质瘤干细胞的p-STAT3表达受到抑制(P < 0.05)，STAT3表达无明显变化，见图3。随着槲皮素浓度的增加，胶质瘤干细胞p-STAT3的表达逐渐降低，表明槲皮素可抑制STAT3的活化，从而抑制STAT通路。 2.4 STAT3敲降抑制胶质瘤干细胞增殖并诱导其凋亡为了研究STAT通路对胶质瘤干细胞增殖和凋亡的作用，向胶质瘤干细胞中转染表达si STAT3的重组质粒，敲降STAT3表达，MTT法检测细胞增殖情况，流式细胞仪检测细胞凋亡情况，Western blot检测STAT3的干扰效果及增殖相关蛋白增殖细胞核抗原、凋亡相关蛋白Bax、Bcl-2、存活素的表达。结果显示，与对照组比较，实验组转染表达si STAT3的重组质粒后，胶质瘤干细胞STAT3表达明显减少(P < 0.05)，表明成功敲降了胶质瘤干细胞中STAT3的表达；细胞增殖受到抑制(P < 0.05)，增殖相关蛋白增殖细胞核抗原表达降低；凋亡细胞数增加(P < 0.05)，凋亡促进蛋白Bax表达升高，凋亡抑制蛋白Bcl-2和存活素表达降低(P < 0.05)，见图4。以上结果表明，STAT通路对胶质瘤干细胞的增殖和凋亡有重要的作用，抑制STAT通路，可导致胶质瘤干细胞增殖受到抑制，诱导胶质瘤干细胞的凋亡。"

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