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

doi:10.3877/cma.j.issn.2095-655X.2024.02.004

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摘要目的探讨人参皂苷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（0.4mg/ml，0.8mg/ml，1.6mg/ml），采用四甲基偶氮唑蓝（MTT）法检测各组细胞24h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0.4mg/ml、0.8mg/ml、1.6mg/ml时分别处理细胞24h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0.8mg/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，0.4mg/ml和0.8mg/ml的Rg3处理实验组细胞出现了凋亡典型特征，1.6mg/ml Rg3处理实验组细胞出现碎片化。Western blot结果显示，0.8mg/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0.8mg/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。

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关键词 ：人参皂苷Rg3, 乳腺肿瘤, 细胞周期蛋白依赖性激酶2, 凋亡, 增殖

Abstract：ObjectiveTo investigate the effect of ginsenoside Rg3 on the activity, apoptosis and proliferation of human breast cancer cell line MDA-MB-231. MethodsHuman breast cancer cell line MDA-MB-231 was cultured in vitro and divided into control group (normal culture) and Rg3 treatment group (0.4mg/ml, 0.8mg/ml, 1.6mg/ml). The metabolic activity of cells in each group within 24h 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. ResultsHuman 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 0.4mg/ml, 0.8mg/ml, and 1.6mg/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 0.8mg/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 0.4mg/ml and 0.8mg/ml showed typical apoptotic characteristics, while the experimental group with Rg3 concentration of 1.6mg/ml showed fragmentation. Western blot results showed that the expression level of CDK2 protein in 0.8mg/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). ConclusionRg3 with a concentration of 0.8mg/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.

Key words： Ginsenoside Rg3 Breast neoplasms Cyclin-dependent kinase 2 Apoptosis Proliferation

收稿日期: 2024-02-24

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）

通讯作者: 随蓓蓓， Email： babe3093@163.com

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]. 中华诊断学电子杂志, 2024, 12(2): 90-94.
Lou Yanwen1, Li Han2, Li Yunhong1, Xu Mengran1, Wei Yanghang1, Sui Beibei1. Effects of ginsenoside Rg3 on metabolic activity and expression of caspase 3 and CDK2 in human breast cancer cells. zhzdx, 2024, 12(2): 90-94.

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