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

doi:10.11958/20250238

Abstract

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

CLC Number:

R737.9

Figures/Tables 6

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

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

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

Fig.3 Formation and histological evaluation of PDOs

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

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

References 32

[1]	SIEGEL R L, GIAQUINTO A N, JEMAL A. Cancer statistics,2024[J]. CA Cancer J Clin, 2024, 74(1):12-49. doi:10.3322/caac.21820.
[2]	RIZZO A, SCHIPILLITI F M, DI COSTANZO F, et al. Discontinuation rate and serious adverse events of chemoimmunotherapy as neoadjuvant treatment for triple-negative breast cancer:a systematic review and meta-analysis[J]. ESMO Open, 2023, 8(6):102198. doi:10.1016/j.esmoop.2023.102198.
[3]	HU M, XU M, CHEN Y, et al. Therapeutic potential of toosendanin:novel applications of an old ascaris repellent as a drug candidate[J]. Biomed Pharmacother, 2023, 167:115541. doi:10.1016/j.biopha.2023.115541.
[4]	LI S, XIONG Q, SHEN Y, et al. Toosendanin:upgrade of an old agent in cancer treatment[J]. Chin J Nat Med, 2024, 22(10):887-899. doi:10.1016/S1875-5364(24)60693-X.
[5]	JIANG Y Z, MA D, SUO C, et al. Genomic and transcriptomic landscape of triple-negative breast cancers:subtypes and treatment strategies[J]. Cancer Cell, 2019, 35(3):428-440.e5. doi:10.1016/j.ccell.2019.02.001.
[6]	TUTT A, GARBER J E, KAUFMAN B, et al. Adjuvant olaparib for patients with BRCA1- or BRCA2-mutated breast cancer[J]. N Engl J Med, 2021, 384(25):2394-2405. doi:10.1056/NEJMoa2105215.
[7]	WU S, YAO X, SUN W, et al. Exploration of poly(ADP-ribose)polymerase inhibitor resistance in the treatment of BRCA1/2-mutated cancer[J]. Genes Chromosomes Cancer, 2024, 63(5):e23243. doi:10.1002/gcc.23243.
[8]	WOOTEN J, MAVINGIRE N, DAMAR K, et al. Triumphs and challenges in exploiting poly(ADP-ribose) polymerase inhibition to combat triple-negative breast cancer[J]. J Cell Physiol, 2023, 238(8):1625-1640. doi:10.1002/jcp.31015.
[9]	LI H, LIU Z Y, WU N, et al. PARP inhibitor resistance:the underlying mechanisms and clinical implications[J]. Mol Cancer, 2020, 19(1):107. doi:10.1186/s12943-020-01227-0.
[10]	KOCAK M, EZAZI ERDI S, JORBA G, et al. Targeting autophagy in disease:established and new strategies[J]. Autophagy, 2022, 18(3):473-495. doi:10.1080/15548627.2021.1936359.
[11]	中国抗癌协会乳腺癌专业委员会,中华医学会肿瘤学分会乳腺肿瘤学组. 中国抗癌协会乳腺癌诊治指南与规范(2024年版)[J]. 中国癌症杂志, 2023, 33(12):1092-1187.
	The Society of Breast Cancer China Anti-Cancer Association, Breast Oncology Group of the Oncology Branch of the Chinese Medical Association. Guidelines for breast cancer diagnosis and treatment by China Anti-cancer Association(2024 edition)[J]. China Oncology, 2023, 33(12):1092-1187. doi:10.19401/j.cnki.1007-3639.2023.12.004.
[12]	QARI A S, MOWAIS A H, ALHARBI S M, et al. Adjuvant and neoadjuvant therapy for breast cancer:a systematic review[J]. Eur J Breast Health, 2024, 20(3):156-166. doi:10.4274/ejbh.galenos.2024.2023-12-16.
[13]	SHI J, LIU F, SONG Y. Progress:targeted therapy,immunotherapy,and new chemotherapy strategies in advanced triple-negative breast cancer[J]. Cancer Manag Res, 2020, 12:9375-9387. doi:10.2147/CMAR.S272685.
[14]	MAO C, GONG L, KANG W. Effect and mechanism of resveratrol on ferroptosis mediated by p53/SLC7A11 in oral squamous cell carcinoma[J]. BMC Oral Health, 2024, 24(1):773. doi:10.1186/s12903-024-04395-3.
[15]	SUI X, HAN X, CHEN P, et al. Baicalin induces apoptosis and suppresses the cell cycle progression of lung cancer cells through downregulating Akt/mTOR signaling pathway[J]. Front Mol Biosci, 2020, 7:602282. doi:10.3389/fmolb.2020.602282.
[16]	GUO Z, ZHANG Y, GONG Y, et al. Antibody functionalized curcuma-derived extracellular vesicles loaded with doxorubicin overcome therapy-induced senescence and enhance chemotherapy[J]. J Control Release, 2025, 379:377-389. doi:10.1016/j.jconrel.2025.01.029.
[17]	IRITI M, KUBINA R, COCHIS A, et al. Rutin,a quercetin glycoside,restores chemosensitivity in human breast cancer cells[J]. Phytother Res, 2017, 31(10):1529-1538. doi:10.1002/ptr.5878.
[18]	ZHANG J, YANG F, MEI X, et al. Toosendanin and isotoosendanin suppress triple-negative breast cancer growth via inducing necrosis,apoptosis and autophagy[J]. Chem Biol Interact, 2022, 351:109739. doi:10.1016/j.cbi.2021.109739.
[19]	WANG G, FAN X Q, LI L, et al. Toosendanin shows potent efficacy against human ovarian cancer through caspase-dependent mitochondrial apoptotic pathway[J]. Am J Chin Med, 2021, 49(7):1757-1772. doi:10.1142/S0192415X2150083X.
[20]	WU Q, SHARMA D. Autophagy and breast cancer:connected in growth,progression,and therapy[J]. Cells, 2023, 12(8):1156. doi:10.3390/cells12081156.
[21]	ZHANG S, DONG Y, CHEN X, et al. Toosendanin,a late-stage autophagy inhibitor,sensitizes triple-negative breast cancer to irinotecan chemotherapy[J]. Chin Med, 2022, 17(1):55. doi:10.1186/s13020-022-00605-8.
[22]	陈芷彦, 伍秋苑, 邓裕华, 等. 类器官技术在乳腺癌研究中的现状及应用[J]. 天津医药, 2024, 52(6):668-672.
	CHEN Z Y, WU Q Y, DENG Y H, et al. Status and application of organoid technology in breast cancer research[J]. Tianjin Med J, 2024, 52(6):668-672. doi:10.11958/20231745.
[23]	JAIN V, SINGH M P, AMARAVADI R K. Recent advances in targeting autophagy in cancer[J]. Trends Pharmacol Sci, 2023, 44(5):290-302. doi:10.1016/j.tips.2023.02.003.
[24]	VISWANATH P, RADOUL M, IZQUIERDO-GARCIA J L, et al. 2-hydroxyglutarate-mediated autophagy of the endoplasmic reticulum leads to an unusual downregulation of phospholipid biosynthesis in mutant IDH1 gliomas[J]. Cancer Res, 2018, 78(9):2290-2304. doi:10.1158/0008-5472.CAN-17-2926.
[25]	BOONE B A, BAHARY N, ZUREIKAT A H, et al. Safety and biologic response of pre-operative autophagy inhibition in combination with gemcitabine in patients with pancreatic adenocarcinoma[J]. Ann Surg Oncol, 2015, 22(13):4402-4410. doi:10.1245/s10434-015-4566-4.
[26]	ZEH H J, BAHARY N, BOONE B A, et al. A randomized phase ii preoperative study of autophagy inhibition with high-dose hydroxychloroquine and gemcitabine/nab-paclitaxel in pancreatic cancer patients[J]. Clin Cancer Res, 2020, 26(13):3126-3134. doi:10.1158/1078-0432.CCR-19-4042.
[27]	KINSEY C G, CAMOLOTTO S A, BOESPFLUG A M, et al. Protective autophagy elicited by RAF→MEK→ERK inhibition suggests a treatment strategy for RAS-driven cancers[J]. Nat Med, 2019, 25(4):620-627. doi:10.1038/s41591-019-0367-9.
[28]	HOU W, XIAO C, ZHOU R, et al. Inhibiting autophagy selectively prunes dysfunctional tumor vessels and optimizes the tumor immune microenvironment[J]. Theranostics, 2025, 15(1):258-276. doi:10.7150/thno.98285.
[29]	MORAND S, STANBERY L, WALTER A, et al. BRCA1/2 Mutation status impact on autophagy and immune response:unheralded target[J]. JNCI Cancer Spectr, 2020, 4(6):pkaa077. doi:10.1093/jncics/pkaa077.
[30]	CAHUZAC M, LANGLOIS P, PÉANT B, et al. Pre-activation of autophagy impacts response to olaparib in prostate cancer cells[J]. Commun Biol, 2022, 5(1):251. doi:10.1038/s42003-022-03210-5.
[31]	XIAO M, YANG J, DONG M, et al. NLRP4 renders pancreatic cancer resistant to olaparib through promotion of the DNA damage response and ROS-induced autophagy[J]. Cell Death Dis, 2024, ,l15(8):620. doi:10.1038/s41419-024-06984-0.
[32]	WANG L H, WEI S, YUAN Y, et al. KPT330 promotes the sensitivity of glioblastoma to olaparib by retaining SQSTM1 in the nucleus and disrupting lysosomal function[J]. Autophagy, 2024, 20(2):295-310. doi:10.1080/15548627.2023.2252301.