Cucurbitacin IIa promotes the immunogenic cell death‑inducing effect of doxorubicin and modulates immune microenvironment in liver cancer

Li,Sujuan; Wang,Sen; Zhang,Anping; Luo,Lixia; Song,Jie; Wei,Guoli; Fang,Zhijun

doi:10.3892/ijo.2024.5625

Cucurbitacin IIa promotes the immunogenic cell death‑inducing effect of doxorubicin and modulates immune microenvironment in liver cancer

Authors:
- Sujuan Li
- Sen Wang
- Anping Zhang
- Lixia Luo
- Jie Song
- Guoli Wei
- Zhijun Fang
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Affiliations: Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, P.R. China
Published online on: February 20, 2024 https://doi.org/10.3892/ijo.2024.5625
Article Number: 37
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The immunogenic cell death (ICD) has aroused great interest in cancer immunotherapy. Doxorubicin (DOX), which can induce ICD, is a widely used chemotherapeutic drug in liver cancer. However, DOX‑induced ICD is not potent enough to initiate a satisfactory immune response. Cucurbitacin IIa (CUIIa), a tetracyclic triterpene, is a biologically active compound present in the Cucurbitaceae family. The present study assessed the effects of the combination of DOX and CUIIa on the viability, colony formation, apoptosis and cell cycle of HepG2 cells. In vivo anticancer effect was performed in mice bearing H22 tumor xenografts. The hallmark expression of ICD was tested using immunofluorescence and an ATP assay kit. The immune microenvironment was analyzed using flow cytometry. The combination of CUIIa and DOX displayed potent apoptosis inducing, cell cycle arresting and in vivo anticancer effects, along with attenuated cardiotoxicity in H22 mice. The combination of DOX and CUIIa also facilitated ICD as manifested by elevated high‑mobility group box 1, calreticulin and ATP secretion. This combination provoked a stronger immune response in H22 mice, including dendritic cell activation, increment of cytotoxic T cells and T helper 1 cells. Moreover, the proportion of immunosuppressive cells including myeloid‑derived suppressor cells, T regulatory cells and M2‑polarized macrophages, decreased. These data suggested that CUIIa is a promising combination partner with DOX for liver cancer treatment, probably via triggering ICD and remolding the immune microenvironment.

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1	Huang J, Hao P, Zhang YL, Deng FX, Deng Q, Hong Y, Wang XW, Wang Y, Li TT, Zhang XG, et al: Discovering multiple transcripts of human hepatocytes using massively parallel signature sequencing (MPSS). BMC Genomics. 8:2072007.
2	Plaz Torres MC, Bodini G, Furnari M, Marabotto E, Zentilin P, Strazzabosco M and Giannini EG: Surveillance for hepatocellular carcinoma in patients with non-alcoholic fatty liver disease: universal or selective? Cancers (Basel). 12:14222020.
3	He T, Wang L, Gou S, Lu L, Liu G, Wang K, Yang Y, Duan Q, Geng W, Zhao P, et al: Enhanced immunogenic cell death and antigen presentation via engineered bifidobacterium bifidum to boost chemo-immunotherapy. ACS Nano. 17:9953–9971. 2023.
4	Lu Q, Huang H, Wang X, Luo L, Xia H, Zhang L, Xu J, Huang Y, Luo X and Luo J: Echinatin inhibits the growth and metastasis of human osteosarcoma cells through Wnt/β-catenin and p38 signaling pathways. Pharmacol Res. 191:1067602023.
5	Vanmeerbeek I, Sprooten J, De Ruysscher D, Tejpar S, Vandenberghe P, Fucikova J, Spisek R, Zitvogel L, Kroemer G, Galluzzi L, et al: Trial watch: Chemotherapy-induced immunogenic cell death in immuno-oncology. Oncoimmunology. 9:17034492020.
6	Obeid M, Tesniere A, Ghiringhelli F, Fimia GM, Apetoh L, Perfettini JL, Castedo M, Mignot G, Panaretakis T, Casares N, et al: Calreticulin exposure dictates the immunogenicity of cancer cell death. Nat Med. 13:54–61. 2007.
7	Green DR, Ferguson T, Zitvogel L and Kroemer G: Immunogenic and tolerogenic cell death. Nat Rev Immunol. 9:353–363. 2009.
8	Casares N, Pequignot MO, Tesniere A, Ghiringhelli F, Roux S, Chaput N, Schmitt E, Hamai A, Hervas-Stubbs S, Obeid M, et al: Caspase-dependent immunogenicity of doxorubicin-induced tumor cell death. J Exp Med. 202:1691–1701. 2005.
9	Birmpilis AI, Paschalis A, Mourkakis A, Christodoulou P, Kostopoulos IV, Antimissari E, Terzoudi G, Georgakilas AG, Armpilia C, Papageorgis P, et al: Immunogenic cell death, DAMPs and prothymosin α as a putative anticancer immune response biomarker. Cells. 11:14152022.
10	Zhai J, Gu X, Liu Y, Hu Y, Jiang Y and Zhang Z: Chemotherapeutic and targeted drugs-induced immunogenic cell death in cancer models and antitumor therapy: An update review. Front Pharmacol. 14:11529342023.
11	Shi Y, Hou X, Yu S, Pan X, Yang M, Hu J and Wang X: Targeted delivery of doxorubicin into tumor cells to decrease the in vivo toxicity of glutathione-sensitive prodrug-poloxamer188-b-polycaprolactone nanoparticles and improve their anti-tumor activities. Colloids Surf B Biointerfaces. 220:1128742022.
12	Wu PJ, Chiou HL, Hsieh YH, Lin CL, Lee HL, Liu IC and Ying TH: Induction of immunogenic cell death effect of licoricidin in cervical cancer cells by enhancing endoplasmic reticulum stress-mediated high mobility group box 1 expression. Environ Toxicol. 38:1641–1650. 2023.
13	Aria H and Rezaei M: Immunogenic cell death inducer peptides: A new approach for cancer therapy, current status and future perspectives. Biomed Pharmacother. 161:1145032023.
14	Lu Y, Sun W, Du J, Fan J and Peng X: Immuno-photodynamic therapy (IPDT): Organic photosensitizers and their application in cancer ablation. JACS Au. 3:682–699. 2023.
15	Shimabukuro-Vornhagen A, Draube A, Liebig TM, Rothe A, Kochanek M and von Bergwelt-Baildon MS: The immunosuppressive factors IL-10, TGF-β, and VEGF do not affect the antigen-presenting function of CD40-activated B cells. J Exp Clin Cancer Res. 31:472012.
16	Fan X, Jin J, Yan L, Liu L, Li Q and Xu Y: The impaired anti-tumoral effect of immune surveillance cells in the immune microenvironment of gastric cancer. Clin Immunol. 219:1085512020.
17	Dai Z, Tang J, Gu Z, Wang Y, Yang Y, Yang Y and Yu C: Eliciting immunogenic cell death via a unitized nanoinducer. Nano Lett. 20:6246–6254. 2020.
18	Wu H, Wei G, Luo L, Li L, Gao Y, Tan X, Wang S, Chang H, Liu Y, Wei Y, et al: Ginsenoside Rg3 nanoparticles with permeation enhancing based chitosan derivatives were encapsulated with doxorubicin by thermosensitive hydrogel and anti-cancer evaluation of peritumoral hydrogel injection combined with PD-L1 antibody. Biomater Res. 26:772022.
19	Yu Z, Guo J, Hu M, Gao Y and Huang L: Icaritin exacerbates mitophagy and synergizes with doxorubicin to induce immunogenic cell death in hepatocellular carcinoma. ACS Nano. 14:4816–4828. 2020.
20	Zeng Y, Wang J, Huang Q, Ren Y, Li T, Zhang X, Yao R and Sun J: Cucurbitacin IIa: A review of phytochemistry and pharmacology. Phytother Res. 35:4155–4170. 2021.
21	Peng Y, Chen T, Luo L, Li L, Cao W, Xu X, Zhang Y, Yue P, Dai X, Ji Z, et al: Isoforskolin and cucurbitacin IIa promote the expression of anti-inflammatory regulatory factor SIGIRR in human macrophages stimulated with Borrelia burgdorferi basic membrane protein A. Int Immunopharmacol. 88:1069142020.
22	Singh N, Krishnakumar S, Kanwar RK, Cheung CH and Kanwar JR: Clinical aspects for survivin: A crucial molecule for targeting drug-resistant cancers. Drug Discov Today. 20:578–587. 2015.
23	Zhang J, Song Y, Liang Y, Zou H, Zuo P, Yan M, Jing S, Li T, Wang Y, Li D, et al: Cucurbitacin IIa interferes with EGFR-MAPK signaling pathway leads to proliferation inhibition in A549 cells. Food Chem Toxicol. 132:1106542019.
24	Boykin C, Zhang G, Chen YH, Zhang RW, Fan XE, Yang WM and Lu Q: Cucurbitacin IIa: a novel class of anti-cancer drug inducing non-reversible actin aggregation and inhibiting survivin independent of JAK2/STAT3 phosphorylation. Br J Cancer. 104:781–789. 2011.
25	Yu K, Yang X, Li Y, Cui X, Liu B and Yao Q: Synthesis of cucurbitacin IIa derivatives with apoptosis-inducing capabilities in human cancer cells. RSC Adv. 10:3872–3881. 2020.
26	Kuang Z, Wu J, Tan Y, Zhu G, Li J and Wu M: MicroRNA in the diagnosis and treatment of doxorubicin-induced cardiotoxicity. Biomolecules. 13:5682023.
27	Yu S, Cai X, Wu C, Liu Y, Zhang J, Gong X, Wang X, Wu X, Zhu T, Mo L, et al: Targeting HSP90-HDAC6 regulating network implicates precision treatment of breast cancer. Int J Biol Sci. 13:505–517. 2017.
28	O'Donohue TJ, Ibáñez G, Coutinho DF, Mauguen A, Siddiquee A, Rosales N, Calder P, Ndengu A, You D, Long M, et al: Translational strategies for repotrectinib in neuroblastoma. Mol Cancer Ther. 20:2189–2197. 2021.
29	Bhagat A and Kleinerman ES: Anthracycline-induced cardiotoxicity: Causes, mechanisms, and prevention. Adv Exp Med Biol. 1257:181–192. 2020.
30	Ma X, Yang S, Zhang T, Wang S, Yang Q, Xiao Y, Shi X, Xue P, Kang Y, Liu G, et al: Bioresponsive immune-booster-based prodrug nanogel for cancer immunotherapy. Acta Pharm Sin B. 12:451–466. 2022.
31	Bao L, Hao C, Wang J, Wang D, Zhao Y, Li Y and Yao W: High-dose cyclophosphamide administration orchestrates phenotypic and functional alterations of immature dendritic cells and regulates Th cell polarization. Front Pharmacol. 11:7752020.
32	Shan Z, Wang H, Zhang Y and Min W: The role of tumor-derived exosomes in the abscopal effect and immunotherapy. Life (Basel). 11:3812021.
33	Deng Z, Zhang M, Zhu T, Zhili N, Liu Z, Xiang R, Zhang W and Xu Y: Dynamic changes in peripheral blood lymphocyte subsets in adult patients with COVID-19. Int J Infect Dis. 98:353–358. 2020.
34	Yu H, Zou W, Mi C, Wang Q, Dai G, Zhang T, Zhang G, Xie K, Wang J and Shi H: Research Note: Expression of T cell-related cytokines in chicken cecal and spleen tissues following Eimeria tenella infection in vivo. Poult Sci. 100:1011612021.
35	Sasidharan Nair V, Saleh R, Toor SM, Taha RZ, Ahmed AA, Kurer MA, Murshed K, Alajez NM, Abu Nada M and Elkord E: Transcriptomic profiling disclosed the role of DNA methylation and histone modifications in tumor-infiltrating myeloid-derived suppressor cell subsets in colorectal cancer. Clin Epigenetics. 12:132020.
36	Wellenstein MD and de Visser KE: Cancer-cell-intrinsic mechanisms shaping the tumor immune landscape. Immunity. 48:399–416. 2018.
37	Si W, Lyu J, Liu Z, Wang C, Huang J, Jiang L and Ma T: Cucurbitacin E inhibits cellular proliferation and enhances the chemo-response in gastric cancer by suppressing AKt activation. J Cancer. 10:5843–5851. 2019.
38	Yin S, Mai Z, Liu C, Xu L and Xia C: Label-free-based quantitative proteomic analysis of the inhibition of cisplatin-resistant ovarian cancer cell proliferation by cucurbitacin B. Phytomedicine. 111:1546692023.
39	Ni K, Lan G, Guo N, Culbert A, Luo T, Wu T, Weichselbaum RR and Lin W: Nanoscale metal-organic frameworks for x-ray activated in situ cancer vaccination. Sci Adv. 6:eabb52232020.
40	Sun D, Zou Y, Song L, Han S, Yang H, Chu D, Dai Y, Ma J, O'Driscoll CM, Yu Z and Guo J: A cyclodextrin-based nanoformulation achieves co-delivery of ginsenoside Rg3 and quercetin for chemo-immunotherapy in colorectal cancer. Acta Pharm Sin B. 12:378–393. 2022.
41	Yang Q, Shi G, Chen X, Lin Y, Cheng L, Jiang Q, Yan X, Jiang M, Li Y, Zhang H, et al: Nanomicelle protects the immune activation effects of Paclitaxel and sensitizes tumors to anti-PD-1 immunotherapy. Theranostics. 10:8382–8399. 2020.
42	Wu Q, Li B, Li J and Sun S, Yuan J and Sun S: Cancer-associated adipocytes as immunomodulators in cancer. Biomark Res. 9:22021.
43	Szczygieł A, Węgierek-Ciura K, Wróblewska A, Mierzejewska J, Rossowska J, Szermer-Olearnik B, Świtalska M, Anger-Góra N, Goszczyński TM and Pajtasz-Piasecka E: Combined therapy with methotrexate nanoconjugate and dendritic cells with downregulated IL-10R expression modulates the tumor microenvironment and enhances the systemic anti-tumor immune response in MC38 murine colon carcinoma. Front Immunol. 14:11553772023.
44	Evgin L and Vile RG: Parking CAR T cells in tumours: Oncolytic viruses as valets or vandals? Cancers (Basel). 13:11062021.
45	Li J, Zhao M, Liang W, Wu S, Wang Z and Wang D: Codelivery of Shikonin and siTGF-β for enhanced triple negative breast cancer chemo-immunotherapy. J Control Release. 342:308–320. 2022.
46	Shao Y, Lo CM, Ling CC, Liu XB, Ng KTP, Chu ACY, Ma YY, Li CX, Fan ST and Man K: Regulatory B cells accelerate hepatocellular carcinoma progression via CD40/CD154 signaling pathway. Cancer Lett. 355:264–272. 2014.
47	Liu M, Zhou J, Liu X, Feng Y, Yang W, Wu F, Cheung OKW, Sun H, Zeng X, Tang W, et al: Targeting monocyte-intrinsic enhancer reprogramming improves immunotherapy efficacy in hepatocellular carcinoma. Gut. 69:365–379. 2020.
48	He F, Furones AR, Landegren N, Fuxe J and Sarhan D: Sex dimorphism in the tumor microenvironment-from bench to bedside and back. Semin Cancer Biol. 86:166–179. 2022.
49	He J, Wang Y, Xu LH, Qiao J, Ouyang DY and He XH: Cucurbitacin IIa induces caspase-3-dependent apoptosis and enhances autophagy in lipopolysaccharide-stimulated RAW 264.7 macrophages. Int Immunopharmacol. 16:27–34. 2013.