放线菌素 D 通过诱导 P53 依赖性细胞凋亡与多柔比星协同作用于三阴性乳腺癌。

Actinomycin D synergizes with Doxorubicin in triple-negative breast cancer by inducing P53-dependent cell apoptosis.

机构信息

The State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Beijing, China.

Key Laboratory of Drug Target Research and Drug Screen, Institute of Materia Medica, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, China.

出版信息

Carcinogenesis. 2024 Apr 12;45(4):262-273. doi: 10.1093/carcin/bgad086.

DOI:10.1093/carcin/bgad086

PMID:37997385

Abstract

OBJECTIVES

There are three major subtypes of breast cancer, ER+, HER2+ and triple-negative breast cancer (TNBC), namely ER-, PR-, HER2-. TNBC is the most aggressive breast cancer with poor prognosis and no target drug up to now. Actinomycin D (ActD) is a bioactive metabolite of marine bacteria that has been reported to have antitumor activity. The aim of study is to investigate whether ActD has a synergetic effect on TNBC with Doxorubicin (Dox), the major chemotherapeutic drug for TNBC, and explore the underlying mechanism.

METHODS

TNBC cell lines HCC1937, MDA-MB-436 and nude mice were used in the study. Drug synergy determination, LDH assay, MMP assay, Hoechst 33342 staining, Flow cytometry, Flexible docking and CESTA assay were carried out. The expression of proteins associated with apoptosis was checked by Western blot and siRNA experiments were performed to investigate the role of P53 and PUMA induced by drugs.

RESULTS

There was much higher apoptosis rate of cells in the ActD + Dox group than that in ActD group or Dox group. Expression of MDM2 and BCL-2 was reduced while expression of P53, PUMA and BAX were increased in the groups treated with ActD + Dox or Dox compared to the control group. Furthermore, P53 siRNA or PUMA siRNA tremendously abrogated the cell apoptosis in the groups treated by ActD, Dox and ActD + Dox. Flexible docking and CESTA showed that ActD can bind MDM2.

CONCLUSIONS

ActD had a synergetic effect on TNBC with Dox via P53-dependent apoptosis and it may be a new choice for treatment of TNBC.

摘要

目的

乳腺癌有三种主要亚型，即雌激素受体阳性（ER+）、人表皮生长因子受体 2 阳性（HER2+）和三阴性乳腺癌（TNBC），即 ER-、PR-、HER2-。TNBC 是最具侵袭性的乳腺癌，预后不良，至今尚无靶向药物。放线菌素 D（ActD）是一种海洋细菌的生物活性代谢物，已被报道具有抗肿瘤活性。本研究旨在探讨 ActD 与多柔比星（Dox）（TNBC 的主要化疗药物）联合应用对 TNBC 是否具有协同作用，并探讨其潜在机制。

方法

本研究采用 TNBC 细胞系 HCC1937、MDA-MB-436 和裸鼠。进行药物协同作用测定、LDH 测定、MMP 测定、Hoechst 33342 染色、流式细胞术、柔性对接和 CESTA 测定。通过 Western blot 检测与细胞凋亡相关的蛋白表达，并通过 siRNA 实验研究药物诱导的 P53 和 PUMA 的作用。

结果

与 ActD 组或 Dox 组相比，ActD+Dox 组细胞的凋亡率明显更高。与对照组相比，ActD+Dox 组或 Dox 组中 MDM2 和 BCL-2 的表达降低，而 P53、PUMA 和 BAX 的表达增加。此外，P53 siRNA 或 PUMA siRNA 极大地阻断了 ActD、Dox 和 ActD+Dox 处理组的细胞凋亡。柔性对接和 CESTA 显示 ActD 可以与 MDM2 结合。