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中华诊断学电子杂志 ›› 2024, Vol. 12 ›› Issue (02) : 90 -94. doi: 10.3877/cma.j.issn.2095-655X.2024.02.004

基础研究

人参皂苷Rg3对人乳腺癌细胞的代谢活性及caspase 3、CDK2表达的影响
娄彦文1, 李涵2, 李运鸿1, 徐萌冉1, 魏洋行1, 随蓓蓓1,()   
  1. 1. 276826 日照,济宁医学院生物科学学院
    2. 272000 济宁市第一人民医院急诊科
  • 收稿日期:2024-02-24 出版日期:2024-05-26
  • 通信作者: 随蓓蓓
  • 基金资助:
    山东省中医药科技面上项目(2021M133); 济宁医学院大学生创新训练计划项目(cx2022100z)

Effects of ginsenoside Rg3 on metabolic activity and expression of caspase 3 and CDK2 in human breast cancer cells

Yanwen Lou1, Han Li2, Yunhong Li1, Mengran Xu1, Yanghang Wei1, Beibei Sui1,()   

  1. 1. School of Biological Science, Jining Medical University, Rizhao 276826, China
    2. Department of Emergency, Jining First People′s Hospital, Jining 272000, China
  • Received:2024-02-24 Published:2024-05-26
  • Corresponding author: Beibei Sui
引用本文:

娄彦文, 李涵, 李运鸿, 徐萌冉, 魏洋行, 随蓓蓓. 人参皂苷Rg3对人乳腺癌细胞的代谢活性及caspase 3、CDK2表达的影响[J/OL]. 中华诊断学电子杂志, 2024, 12(02): 90-94.

Yanwen Lou, Han Li, Yunhong Li, Mengran Xu, Yanghang Wei, Beibei Sui. Effects of ginsenoside Rg3 on metabolic activity and expression of caspase 3 and CDK2 in human breast cancer cells[J/OL]. Chinese Journal of Diagnostics(Electronic Edition), 2024, 12(02): 90-94.

目的

探讨人参皂苷Rg3对人乳腺癌细胞MDA-MB-231代谢活性、凋亡和增殖的影响。

方法

体外培养人乳腺癌细胞MDA-MB-231,分为对照组(正常培养)和Rg3处理实验组(0.4 mg/ml,0.8 mg/ml,1.6 mg/ml),采用四甲基偶氮唑蓝(MTT)法检测各组细胞24 h内的代谢活性;采用凋亡试剂盒检测各组细胞凋亡因子caspase 3的酶活性;采用电镜观察各组细胞形态学变化;采用蛋白质印迹(Western blot)法检测细胞周期蛋白依赖性激酶2(CDK2)蛋白的表达水平。采用t检验比较两组间caspase 3酶活力和CDK2蛋白表达水平。

结果

不同浓度Rg3处理人乳腺癌细胞MDA-MB-231后,细胞活性变弱,具有时间和浓度依赖性。凋亡试剂盒检测发现,Rg3浓度为0.4 mg/ml、0.8 mg/ml、1.6 mg/ml时分别处理细胞24 h后,caspase 3酶活力分别为(116.00±8.71)、(171.00±10.82)、(95.67±5.03),仅Rg3浓度为0.8 mg/ml时,与对照组(105.33±7.51)比较,差异有统计学意义(t=8.64,P<0.01)。电镜观察结果显示,0.4 mg/ml和0.8 mg/ml的Rg3处理实验组细胞出现了凋亡典型特征,1.6 mg/ml Rg3处理实验组细胞出现碎片化。Western blot结果显示,0.8 mg/ml Rg3处理实验组细胞的CDK2蛋白的表达水平(0.76±0.03)与对照组(0.89±0.07)比较显著降低(t=2.84,P<0.05)。

结论

浓度为0.8 mg/ml的Rg3可对人乳腺癌细胞MDA-MB-231代谢活性有明显的抑制作用,促使细胞发生凋亡,降低细胞增殖因子CDK2的表达,对细胞增殖有一定的抑制作用。

Objective

To investigate the effect of ginsenoside Rg3 on the activity, apoptosis and proliferation of human breast cancer cell line MDA-MB-231.

Methods

Human breast cancer cell line MDA-MB-231 was cultured in vitro and divided into control group (normal culture) and Rg3 treatment group (0.4 mg/ml, 0.8 mg/ml, 1.6 mg/ml). The metabolic activity of cells in each group within 24 h was detected by methyl thiazolyl terazolium (MTT) assay. Apoptosis kit was used to detect the activity of caspase 3 in each group. The morphological changes of cells in each group were observed by electron microscopy. The expression level of cyclin-dependent kinase 2 (CDK2) protein was detected by Western blot. Comparison between the enzymatic activity of caspase 3 and the protein expression levels of CDK2 was conducted using t-test.

Results

Human breast cancer cell line MDA-MB-231 was treated with different concentrations of Rg3, and the cell activity was weakened in a time and dose dependent manner. Upon detection with the apoptosis detection kit, it was found that when the concentrations of Rg3 were 0.4 mg/ml, 0.8 mg/ml, and 1.6 mg/ml respectively, the caspase 3 enzyme activities after treating the cells for 24 hours were (116.00±8.71), (171.00±10.82), and (95.67±5.03) respectively. Only at the concentration of 0.8 mg/ml of Rg3, there was a statistically significant difference compared to the control group (105.33±7.51) (t=8.64, P<0.01). The results of electron microscopy showed that the experimental group with Rg3 concentration of 0.4 mg/ml and 0.8 mg/ml showed typical apoptotic characteristics, while the experimental group with Rg3 concentration of 1.6 mg/ml showed fragmentation. Western blot results showed that the expression level of CDK2 protein in 0.8 mg/ml Rg3 treatment group (0.76±0.03) was significantly decreased compared with that in the control group (0.89±0.07) (t=2.84, P<0.05).

Conclusion

Rg3 with a concentration of 0.8 mg/ml can significantly inhibit the metabolic activity of human breast cancer cell line MDA-MB-231, promote cell apoptosis, reduce the expression of cell proliferation factor CDK2, and have a certain inhibitory effect on cell proliferation.

图1 不同人参皂苷Rg3浓度对人乳腺癌细胞吸光值的影响注:随着Rg3浓度升高,细胞吸光值均随时间推移下降,浓度越高,抑制效果越明显
图2 不同人参皂苷Rg3浓度对人乳腺癌细胞caspase 3活力的影响注:A为对照组;B为0.4 mg/ml Rg3处理实验组;C为0.8 mg/ml Rg3处理实验组;D为1.6 mg/ml Rg3处理实验组;处理时间为24 h;0.8 mg/ml Rg3使caspase 3活性上升明显
图3 不同人参皂苷Rg3浓度处理人乳腺癌细胞后透射电镜照片(× 5 000)注:a图为对照组;b图为0.4 mg/ml Rg3处理组;c图为0.8 mg/ml Rg3处理组;d图为1.6 mg/ml Rg3处理组;处理时间为24 h;b、c图显示细胞出现了凋亡典型特征;标尺=2 μm
图4 人参皂苷Rg3对人乳腺癌细胞CDK2蛋白表达的影响注:a图为两组CDK2蛋白表达量比较;b图为两组Western blot图;β-actin为内参;CG为对照组;EG为0.8 mg/ml Rg3处理实验组;处理时间为24 h;CDK2为细胞周期蛋白依赖性激酶2
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