人参皂苷通过下调β-catenin 抑制人骨肉瘤细胞增殖并诱导其凋亡。

Ginsenoside impedes proliferation and induces apoptosis of human osteosarcoma cells by down-regulating β-catenin.

出版信息

Cancer Biomark. 2019;24(4):395-404. doi: 10.3233/CBM-182046.

DOI:10.3233/CBM-182046

Abstract

BACKGROUND

Osteosarcoma (OS) is the most commonly occurred primary bone malignancy with high incident rates among children and adolescents. In pharmacologic treatment, the drug ginsenoside has been shown to exert anticancer effects on several malignant diseases. The purpose of this research was to investigate the effect of ginsenoside on the apoptosis and proliferation of human OS MG-63 and Saos-2 cells by regulating the expression of β-catenin.

METHODS

Human OS MG-63 and Saos-2 cells were assigned into control group, and four groups with treatment by varying concentrations (12.5 μg/mL, 25 μg/mL, 50 μg/mL and 100 μg/mL) of ginsenoside, respectively. Cell growth after treatment was observed through cell slides. The proliferation rate of MG-63 and Saos-2 cells in each group was detected by CCK-8. After cell transfection at 48 h, cell cycle and cell apoptosis were detected by FITC-Annexin V staining and flow cytometry. The protein and mRNA expressions of β-catenin, Cyclin D1, Bcl-2, Bax and cleaved caspase-3 were detected by RT-qPCR and western blot analysis.

RESULTS

With increased exposure and concentration of ginsenoside, the cell density, total cell numbers and the absorbance of MG-63 and Saos-2 cells gradually decreased. FITC-Annexin V and FITC-Annexin V/PI staining demonstrated that the cell proportion at S phase decreased, whereas the total apoptotic rate of MG-63 and Saos-2 cells was increased. Furthermore, RT-qPCR and western blot analysis highlighted a gradual decrease in protein and mRNA expressions of β-catenin, Bcl-2 and Cyclin D1, while an elevation in those of Bax and cleaved caspase-3.

CONCLUSION

The results of this study demonstrate that ginsenoside inhibits proliferation and promotes apoptosis of human OS MG-63 and Saos-2 cells by reducing the expressions of β-catenin, Bcl-2 and Cyclin D1 and increasing the expression of Bax and cleaved caspase-3.

摘要

背景

骨肉瘤（OS）是儿童和青少年中发病率较高的最常见原发性骨恶性肿瘤。在药物治疗中，已证实人参皂苷对几种恶性疾病具有抗癌作用。本研究旨在通过调节β-catenin 的表达来研究人参皂苷对人骨肉瘤 MG-63 和 Saos-2 细胞凋亡和增殖的影响。

方法

将人骨肉瘤 MG-63 和 Saos-2 细胞分为对照组和分别用不同浓度（12.5μg/ml、25μg/ml、50μg/ml 和 100μg/ml）的人参皂苷处理的四组。通过细胞载玻片观察处理后的细胞生长情况。用 CCK-8 检测各组 MG-63 和 Saos-2 细胞的增殖率。转染 48 小时后，用 FITC-Annexin V 染色和流式细胞术检测细胞周期和细胞凋亡。用 RT-qPCR 和 Western blot 分析检测β-catenin、Cyclin D1、Bcl-2、Bax 和 cleaved caspase-3 的蛋白和 mRNA 表达。

结果

随着人参皂苷暴露和浓度的增加，MG-63 和 Saos-2 细胞的细胞密度、总细胞数和吸光度逐渐降低。FITC-Annexin V 和 FITC-Annexin V/PI 染色表明 S 期细胞比例下降，而 MG-63 和 Saos-2 细胞的总凋亡率增加。此外，RT-qPCR 和 Western blot 分析显示β-catenin、Bcl-2 和 Cyclin D1 的蛋白和 mRNA 表达逐渐降低，而 Bax 和 cleaved caspase-3 的表达逐渐升高。