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中国癌症防治杂志

中国癌症防治杂志 ›› 2024, Vol. 16 ›› Issue (3): 302-307.doi: 10.3969/j.issn.1674-5671.2024.03.06

• 基础研究 • 上一篇    下一篇

穿心莲内酯联合多柔比星介导ROS依赖性DNA损伤诱导乳腺癌细胞死亡

  

  1. 广西医科大学附属肿瘤医院乳腺外科；广西医科大学公共卫生学院卫生毒理学教研室；复旦大学公共卫生学院职业卫生与毒理学教研室
  • 出版日期:2024-06-25 发布日期:2024-06-25
  • 通讯作者: 李秋云 E-mail：930248@sr.gxmu.edu.cn
  • 基金资助:
    国家自然科学基金项目（82260479）；广西自然科学基金项目（2022GXNSFDA035061）

Andrographolide combined with doxorubicin induces cell death in breast cancer cell via ROS-dependent DNA damage

  • Online:2024-06-25 Published:2024-06-25

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摘要： 目的 探讨穿心莲内酯联合多柔比星对乳腺癌细胞凋亡的影响及其分子机制。方法 乳腺癌细胞MDA⁃MB⁃231和MCF⁃7分别单独使用多柔比星、穿心莲内酯或联合两药处理后，采用MTT实验、克隆形成实验分别检测细胞增殖能力、克隆形成能力。采用流式细胞术（PI染色和DCFH⁃DA染色）检测细胞死亡情况和细胞ROS水平，Western blot检测乳腺癌细胞的γ⁃H2AX、PARP和α⁃tubulin蛋白表达，添加ROS清除剂NAC进一步检测ROS对细胞DNA的影响。构建乳腺癌裸鼠异位移植瘤模型，单独使用多柔比星、穿心莲内酯或联合两药处理后，检测裸鼠体重、肿瘤体积，通过HE病理染色、Ki⁃67⁃IHC染色分别检测细胞死亡和肿瘤细胞增殖能力，验证联合用药在动物体内的治疗效果。结果 与单独用药组相比，联合用药组乳腺癌细胞的增殖率、存活率和克隆形成能力均降低（均P<0.05），细胞ROS水平升高（P<0.001），γ⁃H2AX蛋白表达升高并诱导PARP蛋白发生裂解；与联合用药组相比，NAC+联合用药组的细胞存活率升高（P<0.05），而细胞ROS水平降低（P<0.001）、γ⁃H2AX蛋白表达减少。在动物实验中，不同处理组的裸鼠体重差异无统计学意义（均P>0.05）；与单独用药组相比，联合用药组裸鼠肿瘤体积较小、肿瘤细胞死亡更多、Ki⁃67表达水平较低（均P<0.05）。结论 穿心莲内酯能增强多柔比星抑制乳腺癌细胞增殖的能力。穿心莲内酯通过增加ROS水平加重细胞DNA损伤可能是诱导乳腺癌细胞死亡的机制。

关键词: 乳腺癌, 穿心莲内酯, 多柔比星, ROS, DNA损伤

Abstract: Objective To investigate the effects of andrographolide combined with doxorubicin on the apoptosis of breast cancer cells and its molecular mechanism. Methods The breast cancer cells MDA⁃MB⁃231 and MCF⁃7 were treated with doxorubicin alone, andrographolide alone, or the combination of both. The proliferation and clonogenesis of the cells were detected by MTT assay and clonogenesis assay, respectively. Cell death and ROS level were detected by flow cytometry (PI staining and DCFH⁃DA staining), and the expression of γ⁃H2AX, PARP and α⁃tubulin in breast cancer cells was detected by Western blot. The effect of ROS on DNA was further detected by adding ROS scavenger NAC. A xenograft model of breast cancer in nude mice was established. After treatment with Doxorubicin, andrographolide or the combination of both, the body weight and tumor volume of nude mice were detected, and the cell death and tumor cell proliferation ability were detected by HE pathological staining and Ki⁃67⁃IHC staining, respectively, to verify the therapeutic effect of the combination treatment. Results Compared with the single drug group, the proliferation, survival rate and clonogenesis ability of breast cancer cells in the combined drug group were decreased (P<0.05), the ROS level was increased (P<0.001), the expression of γ⁃H2AX protein was increased and PARP protein cleavage was induced. Compared with the combined drug group, the survival rate of the cells in the NAC+ combined drug group was increased (P<0.05), while the cell ROS level was decreased (P<0.001), and the expression of γ⁃H2AX protein was reduced. In animal experiments, there was no significant difference in the body weight of nude mice between all of the  groups (all P>0.05); compared with the single drug group, the combined drug group had smaller tumor volume, more tumor cell death, and lower Ki⁃67 expression level in nude mice (all P<0.05). Conclusions Andrographolide can enhance the ability of doxorubicin and inhibit the proliferation of breast cancer cells. Andrographolide aggravates cell DNA damage by increasing ROS levels, which may be the mechanism of inducing breast cancer cell death. 

Key words: Breast cancer, Andrographolide, Doxorubicin, ROS, DNA Damage

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  • 引用本文

    谢昌全, 何凤仪, 卢国栋, 李秋云. 穿心莲内酯联合多柔比星介导ROS依赖性DNA损伤诱导乳腺癌细胞死亡[J]. 中国癌症防治杂志, 2024, 16(3): 302-307.

    XIE Changquan, HE Fengyi, LU Guodong, LI Qiuyun. Andrographolide combined with doxorubicin induces cell death in breast cancer cell via ROS-dependent DNA damage[J]. Chinese Journal of Oncology Prevention and Treatment, 2024, 16(3): 302-307.