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中华乳腺病杂志(电子版) ›› 2025, Vol. 19 ›› Issue (01) : 27 -32. doi: 10.3877/cma.j.issn.1674-0807.2025.01.005

论著

罗汉果醇对人乳腺癌细胞自噬和凋亡的影响
王峰1,(), 曲更宝1, 王文彦1, 代艳亭1   
  1. 1.100050 北京,首都医科大学附属北京天坛医院普通外科
  • 收稿日期:2024-09-14 出版日期:2025-02-01
  • 通信作者: 王峰
  • 基金资助:
    北京市优秀人才培养资助青年骨干项目(2016000021469G211)

Effect of mogroside on autophagy and apoptosis in human breast cancer cells

Feng Wang1,(), Gengbao Qu1, Wenyan Wang1, Yanting Dai1   

  1. 1.Department of General Surgery, Beijing Tiantan Hospital Affiliated to Capital Medical University, Beijing 100050, China
  • Received:2024-09-14 Published:2025-02-01
  • Corresponding author: Feng Wang
引用本文:

王峰, 曲更宝, 王文彦, 代艳亭. 罗汉果醇对人乳腺癌细胞自噬和凋亡的影响[J/OL]. 中华乳腺病杂志(电子版), 2025, 19(01): 27-32.

Feng Wang, Gengbao Qu, Wenyan Wang, Yanting Dai. Effect of mogroside on autophagy and apoptosis in human breast cancer cells[J/OL]. Chinese Journal of Breast Disease(Electronic Edition), 2025, 19(01): 27-32.

目的

探讨罗汉果醇(MO)对人乳腺癌细胞自噬和凋亡的影响。

方法

选择对数生长期MDA-MB-231 和SK-BR-3 细胞随机分为对照组(C 组)、不同浓度MO 处理组(加入10、20、40、80 μmol/L的MO)。 采用MTT 法检测细胞增殖率,采用实时细胞电子传感系统检测细胞倍增时间,采用吖啶橙(AO)染色检测细胞自噬,采用细胞分析仪检测细胞凋亡水平,采用Western blot 检测自噬蛋白[微管相关蛋白1 轻链3β(LC3β)和p62]及凋亡相关蛋白[Bcl-2 相关X 蛋白(BAX)、B 细胞淋巴瘤2 蛋白(Bcl-2)]及磷脂酰肌醇3 激酶α 亚基(PI3KCA)、蛋白激酶B1(AKT1)的表达水平。 细胞增殖率、倍增时间、自噬体相对含量、细胞凋亡率、蛋白表达水平的组间比较均采用单因素方差分析,进一步两两比较采用LSD-t 检验。

结果

MDA-MB-231 和SK-BR-3 细胞增殖率和倍增时间在C 组、不同浓度MO 处理组之间的组间比较,差异均有统计学意义(F=102.570、110.256、85.238、79.014,P 均<0.001),不同浓度MO 处理组的细胞增殖率均低于C 组,倍增时间均高于C 组(P 均<0.050)。 不同浓度MO 处理组自噬体相对含量均高于C 组(P 均<0.050)。 随着MO 浓度升高,MDA-MB-231 和SK-BR-3 细胞自噬体相对含量均增加,两两比较差异均有统计学意义(P 均<0.050)。 MDA-MB-231 和SK-BR-3 细胞凋亡率多组比较,差异均有统计学意义(F=161.327,151.304;P 均<0.001),MO 组细胞凋亡率均高于C 组(P 均<0.050);随着MO 浓度升高,MDA-MB-231 和SK-BR-3 细胞凋亡率均增加,两两比较差异均有统计学意义(P 均<0.050)。不同组别MDA-MB-231 和SK-BR-3 细胞中LC3β、p62、BAX、Bcl-2、PI3KCA 和AKT1 蛋白表达水平比较,差异均有统计学意义(F=97.150,101.254,81.578,83.336,78.541,85.233,68.435,71.332,163.576,152.673,138.107,143.188;P 均<0.001),MO 各浓度LC3β、BAX、PI3KCA、AKT1 蛋白表达均高于C 组,p62、Bcl-2蛋白表达水平均低于C 组(P 均<0.050);随着MO 浓度升高,BAX、PI3KCA、AKT1 蛋白表达水平升高,Bcl-2 蛋白表达水平降低,两两比较差异均有统计学意义(P 均<0.050)。

结论

MO 是潜在的三阴性乳腺癌和HER-2 阳性乳腺癌抑制剂,其作用机制可能为靶向PI3K/AKT 诱导乳腺癌细胞自噬和凋亡。

Objective

To explore the effect of mogroside (MO) on autophagy and apoptosis in human breast cancer cells.

Methods

MDA-MB-231 and SK-BR-3 cells in the logarithmic growth phase were randomly divided into control group (C group) and treatment groups (MO-treated groups,treated with 10, 20,40 and 80 μmol/L of MO, respectively). The MTT assay was employed to evaluate the cell proliferation rate,and a real-time cell electronic sensing system was utilized to measure the cell doubling time. Acridine orange(AO) staining was used to detect autophagy and a flow cytometer was applied to analyze apoptosis. Western blotting was performed to determine the expression levels of autophagy-related proteins (LC3β, p62),apoptosis-related proteins (BAX, Bcl-2), as well as PI3KCA and AKT1. One-way analysis of variance was used to compare the cell proliferation rate, doubling time, relative autophagosome content, apoptosis rate, and protein expression among groups, and pairwise comparisons were carried out using the LSD-t test.

Results

There were significant differences in the proliferation rate and doubling time of MDA-MB-231 and SK-BR-3 cells between the C group and different MO-treated groups (F=102.570,110.256,85.238,79.014;P<0.001). Specifically, the cell proliferation rates in the MO-treated groups were significantly lower than those in the C group, while the doubling times were significantly longer (P<0.050). The relative autophagosome content in all MO-treated groups was significantly higher than that in the C group (P<0.050), and it increased with the elevation of MO concentrations (P<0.050 for all pairwise comparisons). Significant differences in apoptosis rates were also noted among groups (F=161.327,151.304;P<0.001). The apoptosis rates in all MOtreated groups were significantly higher than those in the C group, and they increased as the MO concentrations rose (P<0.050 for all pairwise comparisons). Moreover, significant differences were detected in the expression levels of LC3β, p62, BAX, Bcl-2, PI3KCA, and AKT1 in MDA-MB-231 and SK-BR-3 cells among groups(F=97.150,101.254,81.578,83.336,78.541,85.233,68.435,71.332,163.576,152.673,138.107,143.188;P<0.001). The expression levels of LC3β, BAX, PI3KCA, and AKT1 in the MO-treated groups were significantly higher than those in the C group, while the expression levels of p62 and Bcl-2 were significantly lower (P<0.050). With the increase of MO concentrations,the expression levels of BAX,PI3KCA,and AKT1 showed an upward trend, and the Bcl-2 expression showed a downward trend, with all pairwise comparisons being statistically significant (P<0.050).

Conclusions

MO is a potential inhibitor of triple-negative breast cancer and HER-2-positive breast cancer. Its mechanism might be related to targeting the PI3K/AKT signaling pathway to induce autophagy and apoptosis in breast cancer cells.

表1 罗汉果醇对乳腺癌细胞增殖和倍增时间的影响(¯x±s
图1 吖啶橙染色检测MDA-MB-231 细胞自噬水平 a、b、c、d、e 分别表示对照组、10 μmol/L、20 μmol/L、40 μmol/L、80 μmol/L罗汉果醇处理组 注:对照组细胞以绿色荧光为主,胞质均匀;经不同浓度的罗汉果醇处理后,胞质内红黄色荧光逐渐增强
表2 罗汉果醇对乳腺癌细胞自噬水平的影响(¯±s,%)
表3 罗汉果醇对乳腺癌细胞凋亡率的影响(¯x±s,%)
表4 罗汉果醇对乳腺癌细胞自噬相关蛋白表达水平的影响(¯x±s
组别 样本数 LC3β p62 BAX
MDA-MB-231 SK-BR-3 MDA-MB-231 SK-BR-3 MDA-MB-231 SK-BR-3
对照组 6 1.03±0.14 0.99±0.09 0.99±0.09 1.01±0.08 0.97±0.11 0.98±0.13
10μmol/L处理组 6 1.22±0.27a 1.19±0.11a 0.82±0.07a 0.84±0.09a 1.18±0.14a 1.23±0.09a
20μmol/L处理组 6 1.45±0.21a、b 1.52±0.18a、b 0.66±0.11a、b 0.63±0.12a、b 1.39±0.21a、b 1.44±0.16a、b
40μmol/L处理组 6 1.69±0.18a、b、c 1.72±0.16a、b、c 0.54±0.08a、b、c 0.55±0.07a、b、c 1.68±0.23a、b、c 1.64±0.18a、b、c
80μmol/L处理组 6 1.99±0.20a、b、c、d 2.03±0.15a、b、c、d 0.43±0.06a、b、c、d 0.40±0.08a、b、c、d 2.13±0.27a、b、c、d 2.08±0.34a、b、c、d
F 97.150 101.254 81.578 83.336 78.541 85.233
P <0.001 <0.001 <0.001 <0.001 <0.001 <0.001
组别 样本数 Bcl-2 PI3KCA AKT1
MDA-MB-231 SK-BR-3 MDA-MB-231 SK-BR-3 MDA-MB-231 SK-BR-3
对照组 6 0.99±0.06 1.01±0.09 0.98±0.06 1.00±0.05 0.99±0.07 1.03±0.07
10μmol/L处理组 6 0.81±0.09a 0.82±0.05a 1.42±0.13a 1.50±0.21a 1.51±0.18a 1.56±0.20a
20μmol/L处理组 6 0.68±0.08a、b 0.70±0.06a、b 1.75±0.24a、b 1.76±0.31a、b 1.87±0.14a、b 1.94±0.23a、b
40μmol/L处理组 6 0.55±0.07a、b、c 0.52±0.09a、b、c 2.11±0.28a、b、c 2.07±0.26a、b、c 2.33±0.23a、b、c 2.29±0.21a、b、c
80μmol/L处理组 6 0.41±0.08a、b、c、d 0.39±0.07a、b、c、d 3.06±0.38a、b、c、d 2.98±0.41a、b、c、d 3.15±0.19a、b、c、d 3.09±0.25a、b、c、d
F 68.435 71.332 163.576 152.673 138.107 143.188
P <0.001 <0.001 <0.001 <0.001 <0.001 <0.001
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