川楝素联合奥拉帕尼在三阴性乳腺癌中的抗肿瘤机制研究

doi:10.11958/20250238

天津医药 ›› 2025, Vol. 53 ›› Issue (9): 897-902.doi: 10.11958/20250238

• 细胞与分子生物学 • 下一篇

川楝素联合奥拉帕尼在三阴性乳腺癌中的抗肿瘤机制研究

黄慧琦¹(), 伍秋苑¹^,², 张昆¹, 李佩贤³, 熊亚明³, 叶国麟¹, 周丹¹^,²^,^△()

1 佛山市第一人民医院乳腺外科（邮编528000）
2 广东医科大学第一临床医学院
3 佛山市第一人民医院转化医学研究院

收稿日期:2025-01-15 修回日期:2025-05-18 出版日期:2025-09-15 发布日期:2025-09-16
通讯作者: ^△E-mail：zhoudanms@hotmail.com
作者简介:黄慧琦（1993），女，主治医师，主要从事乳腺良恶性肿瘤的诊疗方面研究。E-mail：hhqi2661@fsyyy.com
基金资助:
广东省中医药局科研课题(20231324);广东省基础与应用基础研究基金(2022A1515140091);佛山市登峰计划项目(2020B18);广东省医学科学技术研究基金(A2019329)

Research on the anti-tumor mechanism of toosendanin combined with olaparib in triple negative breast cancer

HUANG Huiqi¹(), WU Qiuyuan¹^,², ZHANG Kun¹, LI Peixian³, XIONG Yaming³, YE Guolin¹, ZHOU Dan¹^,²^,^△()

1 Department of Breast Surgery, Foshan First People's Hospital, Foshan 528000, China
2 the First School of Clinical Medicine, Guangdong Medical University
3 Foshan First People's Hospital Translational Medicine Research Institute

Received:2025-01-15 Revised:2025-05-18 Published:2025-09-15 Online:2025-09-16
Contact: ^△E-mail: zhoudanms@hotmail.com

黄慧琦, 伍秋苑, 张昆, 李佩贤, 熊亚明, 叶国麟, 周丹. 川楝素联合奥拉帕尼在三阴性乳腺癌中的抗肿瘤机制研究[J]. 天津医药, 2025, 53(9): 897-902.

HUANG Huiqi, WU Qiuyuan, ZHANG Kun, LI Peixian, XIONG Yaming, YE Guolin, ZHOU Dan. Research on the anti-tumor mechanism of toosendanin combined with olaparib in triple negative breast cancer[J]. Tianjin Medical Journal, 2025, 53(9): 897-902.

摘要/Abstract

摘要：

目的探讨川楝素（TSN）联合奥拉帕尼在三阴性乳腺癌（TNBC）中的作用机制。方法体外培养人TNBC细胞系MDA-MB-231，利用0.1 μmol/L TSN和0.5、1.0、5.0 μmol/L奥拉帕尼单独或共同处理，评估TSN联合奥拉帕尼对MDA-MB-231自噬水平及细胞活力的影响。选取4例TNBC患者的新辅助化疗后肿瘤残留手术标本，建立患者来源类器官（PDOs）模型，分为5 μmol/L、10 μmol/L奥拉帕尼组，0.05 μmol/L TSN组和TSN+奥拉帕尼组（0.1 μmol/L TSN分别与5、10 μmol/L奥拉帕尼联合处理）。检测TSN联合奥拉帕尼在TNBC患者中的药物敏感性，验证TSN联合奥拉帕尼是否能够在PDOs模型中发挥肿瘤杀伤作用。结果奥拉帕尼在MDA-MB-231细胞系中诱导自噬，相比单独使用奥拉帕尼，TSN和5.0 μmol/L奥拉帕尼联合使用可抑制MDA-MB-231细胞的增殖（P<0.01）。在TNBC的PDOs模型中，与单用奥拉帕尼相比，加用TSN后细胞死亡率明显升高（P<0.01）。结论 TSN联合奥拉帕尼的肿瘤杀伤作用优于单独使用奥拉帕尼，其机制可能与抑制自噬有关。

关键词: 三阴性乳腺癌, 川楝素, 自噬, 类器官, 抗肿瘤药, 植物, 奥拉帕尼, 协同作用

Abstract:

Objective To investigate the anti-tumor mechanism of natural compound toosendanin (TSN) combined with olaparib in triple-negative breast cancer (TNBC). Methods Human TNBC cell line MDA-MB-231 was cultured in vitro. Effects of TSN combined with olaparib on autophagy levels and cell viability in MDA-MB-231 cells were evaluated using 0.5, 1.0, and 5.0 μmol/L olaparib alone or in combination. Surgical specimens from four TNBC patients who had residual tumors after neoadjuvant chemotherapy were selected to establish patient-derived organoid (PDO) models. The drug sensitivity of TSN combined with olaparib in TNBC patients was detected. Whether TSN combined with olaparib can exert autophagy inhibitory effects and tumor-killing effects in organoid model was verified. Results Olaparib induced autophagy in MDA-MB-231 cell line, and the combination of TSN and olaparib inhibited the proliferation of MDA-MB-231 cells (P<0.01). In the TNBC PDOs model, the therapeutic effect of olaparib combined with TSN can significantly reduce the proliferation ability of tumor cells compared with olaparib alone. Conclusion The tumor-killing effect of TSN combined with olaparib is superior to that of olaparib alone, and the mechanism may be related to autophagy inhibition.

Key words: triple negative breast neoplasms, toosendanin, autophagy, organoids, antineoplastic agents, phytogenic, olaparib, synergistic effect

中图分类号:

R737.9

图/表 6

表1 4例TNBC患者临床信息

Tab.1 The clinical information of four patients with TNBC

编号	年龄/岁	性别	病理类型	TNM分期	BRCA突变
BC-1	48	女	左乳浸润性癌	pT₂N₁M₀	无异常突变
BC-2	52	女	右乳浸润性癌	pT₃N₁M₀	无异常突变
BC-3	56	女	左乳浸润性癌	pT₄N₁M₀	BRCA-2突变
BC-4	43	女	右乳浸润性癌	pT₂N₁M₀	BRCA-1突变

图1 奥拉帕尼对TNBC细胞自噬流的影响比例尺：200 μm，红色点状结构（mRFP标记）表示自噬体已与溶酶体融合为成熟自噬溶酶体，提示自噬流完成，黄色点状结构代表当自噬小体尚未与溶酶体融合时，在中性环境中GFP保留其绿色荧光叠加mRFP后显示为黄色。

Fig.1 Effect of olaparib on autophagic flux in TNBC cells

图2 奥拉帕尼联合TSN对MDA-MB-231细胞活力的影响 A：Control组；B：0.1 μmol/L TSN组；C：0.5 μmol/L奥拉帕尼组；D：0.1 μmol/L TSN+0.5 μmol/L奥拉帕尼组；E：1.0 μmol/L奥拉帕尼组；F：0.1 μmol/L TSN+1.0 μmol/L奥拉帕尼组；G：5.0 μmol/L奥拉帕尼组；H：0.1 μmol/L TSN+5.0 μmol/L奥拉帕尼组。

Fig.2 The effect of olaparib combined with TSN on the viability of MDA-MB-231 cells

图3 PDOs的形成与组织学评估 A：组织标本切片HE染色（×20）；B：3D原代培养类器官HE染色（×20）；C：3D原代培养48 h后显微镜下可见细胞增殖分裂（×20）；D：原代培养7 d后类器官（×20）。

Fig.3 Formation and histological evaluation of PDOs

图4 TSN联合不同浓度奥拉帕尼处理后PDOs的细胞活力变化（×40）

Fig.4 Changes in cell viability of PDOs after treatment with TSN combined with different concentrations of olaparib (×40)

图5 不同浓度奥拉帕尼联合TSN处理后PDOs的细胞死亡率比较 A：对照组；B：5 μmol/L奥拉帕尼组；C：10 μmol/L奥拉帕尼组；D：0.05 μmol/L TSN组；E：0.05 μmol/L TSN+5 μmol/L奥拉帕尼组；F：0.05 μmol/L TSN+10 μmol/L奥拉帕尼组。

Fig.5 Comparison of cell mortality rates of PDOs after treatment with different concentrations of olaparib combined with TSN

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川楝素联合奥拉帕尼在三阴性乳腺癌中的抗肿瘤机制研究

Research on the anti-tumor mechanism of toosendanin combined with olaparib in triple negative breast cancer

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