Antineoplastic effects of icaritin: molecular mechanisms and applications

Abstract

Icaritin (ICA) is a prenylflavonoid natural product extracted from plants of the Epimedium genus. The approval of ICA softgel capsules as a class 1.2 traditional Chinese medicine (TCM) innovative drug represents a major advancement offering a novel therapeutic approach for patients with advanced hepatocellular carcinoma (HCC). Class 1.2 TCM innovative drugs generally denote extracts and formulations derived from single plants, animals, minerals, or other substances. ICA exhibits diverse pharmacological effects, encompassing anti-inflammatory, immune-regulatory, anti-oxidation, anti-osteoporosis, anti-depression, and notably anti-cancer properties. This review presents a comprehensive overview of the molecular mechanisms underlying the anti-cancer properties of ICA, and further highlights recent progress in use of ICA in cancer research, discusses present challenges, and examines potential opportunities for ICA application and further development.

Keywords

traditional Chinese medicine / icaritin / hepatocellular carcinoma / anti-tumor / molecular mechanism / application

References

[1] Tsimberidou AM, Fountzilas E, Nikanjam M, Kurzrock R: Review of Precision Cancer Medicine: Evolution of the Treatment Paradigm. Cancer Treatment Review 2020, 86: 102019.

[2] Lee C, Raffaghello L, Longo VD: Starvation, Detoxification, and Multidrug Resistance in Cancer Therapy. Drug Resistance Updates: Reviews and Commentaries in Antimicrobial and Anticancer Chemotherapy 2012, 15: 114-122.

[3] Li K, Xiao K, Zhu S, Wang Y, Wang W: Chinese Herbal Medicine for Primary Liver Cancer Therapy: Perspectives and Challenges. Frontiers in Pharmacology 2022, 13: 889799.

[4] Vogel A, Meyer T, Sapisochin G, Salem R, Saborowski A: Hepatocellular Carcinoma. Lancet 2022, 400: 1345-1362.

[5] Pang X, Yin SS, Yu HY, Zhang Y, Wang T, Hu LM, et al.: Prenylated Flavonoids and Dihydrophenanthrenes from the Leaves of Epimedium Brevicornu and Their Cytotoxicity Against HepG2 Cells. Natural Product Research 2018, 32: 2253-2259.

[6] Anastasiou D, Poulogiannis G, Asara JM, Boxer MB, Jiang JK, Shen M, et al.: Inhibition of Pyruvate Kinase M2 by Reactive Oxygen Species Contributes to Cellular Antioxidant Responses. Science 2011, 334: 1278-1283.

[7] Qin L, Han T, Zhang Q, Cao D, Nian H, Rahman K, et al.: Antiosteoporotic Chemical Constituents from Er-Xian Decoction, a Traditional Chinese Herbal Formula. Journal of Ethnopharmacology 2008, 118: 271-279.

[8] Wu B, Feng JY, Yu LM, Wang YC, Chen YQ, Wei Y, et al.: Icariin Protects Cardiomyocytes Against Ischaemia/Reperfusion Injury by Attenuating Sirtuin 1-Dependent Mitochondrial Oxidative Damage. British Journal of Pharmacology 2018, 175: 4137-4153.

[9] Zhu T, Zhang F, Li H, He Y, Zhang G, Huang N, et al.: Long-term Icariin Treatment Ameliorates Cognitive Deficits via CD4(+) T Cell-Mediated Immuno-Inflammatory Responses in APP/PS1 Mice. Clinical Interventions in Aging 2019, 14: 817-826.

[10] Yang XJ, Xi YM, Li ZJ: Icaritin: A Novel Natural Candidate for Hematological Malignancies Therapy. BioMed Research International 2019, 2019: 4860268.

[11] Tan HL, Chan KG, Pusparajah P, Saokaew S, Duangjai A, Lee LH, et al.: Anti-Cancer Properties of the Naturally Occurring Aphrodisiacs: Icariin and Its Derivatives. Frontier in Pharmacology 2016, 7: 191.

[12] Pitt JM, Marabelle A, Eggermont A, Soria JC, Kroemer G, Zitvogel L: Targeting the Tumor Microenvironment: Removing Obstruction to Anticancer Immune Responses and Immunotherapy. Annals of Oncology: Official Journal of the European Society for Medical Oncology 2016, 27: 1482-1492.

[13] Pistritto G, Trisciuoglio D, Ceci C, Garufi A, D’Orazi G: Apoptosis as Anticancer Mechanism: Function and Dysfunction of its Modulators and Targeted Therapeutic Strategies. Aging (Albany NY) 2016, 8: 603-619.

[14] Fulda S: Targeting Apoptosis for Anticancer Therapy. Seminars in Cancer Biology 2015, 31: 84-88.

[15] Wong RS: Apoptosis in Cancer: From Pathogenesis to Treatment. Journal of Experimental & Clinical Cancer Research 2011, 30: 87.

[16] Metrailler-Ruchonnet I, Pagano A, Carnesecchi S, Ody C, Donati Y, Barazzone Argiroffo C: Bcl-2 Protects Against Hyperoxia-Induced Apoptosis Through Inhibition of the Mitochondria-Dependent Pathway. Free Radical Biology & Medicine 2007, 42: 1062-1074.

[17] Yao Y, Huang C, Li ZF, Wang AY, Liu LY, Zhao XG, et al.: Exogenous Phosphatidylethanolamine Induces Apoptosis of Human Hepatoma HepG2 Cells via the bcl-2/Bax Pathway. World Journal of Gastroenterology 2009, 15: 1751-1758.

[18] Li H, Liu Y, Jiang W, Xue J, Cheng Y, Wang J, et al.: Icaritin Promotes Apoptosis and Inhibits Proliferation by Down-Regulating AFP Gene Expression in Hepatocellular Carcinoma. BMC Cancer 2021, 21: 318.

[19] He J, Wang Y, Duan F, Jiang H, Chen MF, Tang SY: Icaritin Induces Apoptosis of HepG2 Cells via the JNK1 Signaling Pathway Independent of the Estrogen Receptor. Planta Medica 2010, 76: 1834-1839.

[20] Tong JS, Zhang QH, Huang X, Fu XQ, Qi ST, Wang YP, et al.: Icaritin Causes Sustained ERK1/2 Activation and Induces Apoptosis in Human Endometrial Cancer Cells. PLoS One 2011, 6: e16781.

[21] Lu PH, Chen MB, Liu YY, Wu MH, Li WT, Wei MX, et al.: Identification of Sphingosine Kinase 1 (SphK1) as a Primary Target of Icaritin in Hepatocellular Carcinoma Cells. Oncotarget 2017, 8: 22800-22810.

[22] Sun L, Peng Q, Qu L, Gong L, Si J: Anticancer Agent Icaritin Induces Apoptosis Through Caspase-Dependent Pathways in Human Hepatocellular Carcinoma Cells. Molecular Medicine Reports 2015, 11: 3094-3100.

[23] Zheng Q, Liu WW, Li B, Chen HJ, Zhu WS, Yang GX, et al.: Anticancer Effect of Icaritin on Human Lung Cancer Cells Through Inducing S Phase Cell Cycle Arrest and Apoptosis. Journal of Huazhong University of Science and Technology. Medical Science 2014, 34: 497-503.

[24] Zhao X, Lin Y, Jiang B, Yin J, Lu C, Wang J, et al.: Icaritin Inhibits Lung Cancer-Induced Osteoclastogenesis by Suppressing the Expression of IL-6 and TNF-a and Through AMPK/mTOR Signaling Pathway. Anticancer Drugs 2020, 31: 1004-1011.

[25] Gao L, Chen M, Ouyang Y, Li R, Zhang X, Gao X, et al.: Icaritin Induces Ovarian Cancer Cell Apoptosis Through Activation of p53 and Inhibition of Akt/mTOR Pathway. Life Sciences 2018, 202: 188-194.

[26] Hou KM, Li QR, Yang Q, Xu K, Chen R: [Effect of Icaritin on Proliferation and Apoptosis of Human Ovarian Cancer Cells SKOV3]. Zhongguo Zhong Yao Za Zhi 2021, 46: 183-189.

[27] Chen X, Song L, Hou Y, Li F: Reactive Oxygen Species Induced by Icaritin Promote DNA Strand Breaks and Apoptosis in Human Cervical Cancer Cells. Oncology Reports 2019, 41: 765-778.

[28] Yang C, Jin YY, Mei J, Hu D, Jiao X, Che HL, et al.: Identification of Icaritin Derivative IC2 as an SCD-1 Inhibitor with Anti-Breast Cancer Properties Through Induction of Cell Apoptosis. Cancer Cell International 2022, 22: 202.

[29] Yang JG, Lu R, Ye XJ, Zhang J, Tan YQ, Zhou G: Icaritin Reduces Oral Squamous Cell Carcinoma Progression via the Inhibition of STAT3 Signaling. International Journal of Molecular Sciences 2017, 18: 132.

[30] Liu Y, Shi L, Liu Y, Li P, Jiang G, Gao X, et al.: Activation of PPARγ Mediates Icaritin-Induced Cell Cycle Arrest and Apoptosis in Glioblastoma Multiforme. Biomedicine & Pharmacotherapy 2018, 100: 358-366.

[31] Han H, Xu B, Hou P, Jiang C, Liu L, Tang M, et al.: Icaritin Sensitizes Human Glioblastoma Cells to TRAIL-Induced Apoptosis. Cell Biochemistry and Biophysics 2015, 72: 533-542.

[32] Zhu J, Li Z, Zhang G, Meng K, Kuang W, Li J, et al.: Icaritin Shows Potent Anti-Leukemia Activity on Chronic Myeloid Leukemia In Vitro and In Vivo by Regulating MAPK/ERK/JNK and JAK2/STAT3/AKT Signalings. PLoS One 2011, 6: e23720.

[33] Li Q, Huai L, Zhang C, Wang C, Jia Y, Chen Y, et al.: Icaritin Induces AML Cell Apoptosis via the MAPK/ERK and PI3K/AKT Signal Pathways. International Journal of Hematology 2013, 97: 617-623.

[34] Zhu S, Wang Z, Li Z, Peng H, Luo Y, Deng M, et al.: Icaritin Suppresses Multiple Myeloma, by Inhibiting IL-6/JAK2/STAT3. Oncotarget 2015, 6: 10460-10472.

[35] Li ZJ, Yao C, Liu SF, Chen L, Xi YM, Zhang W, et al.: Cytotoxic Effect of Icaritin and its Mechanisms in Inducing Apoptosis in Human Burkitt Lymphoma Cell Line. BioMed Research International 2014, 2014: 391512.

[36] Wu T, Wang S, Wu J, Lin Z, Sui X, Xu X, et al.: Icaritin Induces Lytic Cytotoxicity in Extranodal NK/T-Cell Lymphoma. Journal of Experimental & Clinical Cancer Research 2015, 34: 17.

[37] Lu X, Xue B, Zhang T, Zhou X, Zhang Y: Down-Regulation of MicroRNA-10a Mediates the Anti-Tumor Effect of Icaritin in A549 Cells via the PTEN/AKT and ERK Pathway. General Physiology and Biophysics 2019, 38: 525-533.

[38] Jin L, Miao J, Liu Y, Li X, Jie Y, Niu Q, et al.: Icaritin Induces Mitochondrial Apoptosis by Up-Regulating miR-124 in Human Oral Squamous Cell Carcinoma Cells. Biomedicine & Pharmacotherapy 2017, 85: 287-295.

[39] Li S, Priceman SJ, Xin H, Zhang W, Deng J, Liu Y, et al.: Icaritin Inhibits JAK/STAT3 Signaling and Growth of Renal Cell Carcinoma. PLoS One 2013, 8: e81657.

[40] Hu J, Zhu W, Wei B, Wen H, Mao S, Xu H, et al.: Antitumoral Action of Icaritin in LNCaP Prostate Cancer Cells by Regulating PEA3/HER2/AR Signaling. Anticancer Drugs 2016, 27: 944-952.

[41] Wu J, Du J, Fu X, Liu B, Cao H, Li T, et al.: Iciartin, a Novel FASN Inhibitor, Exerts Anti-Melanoma Activities Through IGF-1R/STAT3 Signaling. Oncotarget 2016, 7: 51251-51269.

[42] Wang C, Li Y, Yan S, Wang H, Shao X, Xiao M, et al.: Interactome Analysis Reveals that lncRNA HULC Promotes Aerobic Glycolysis Through LDHA and PKM2. Nature Communications 2020, 11: 3162.

[43] Debnath J, Gammoh N, Ryan KM: Autophagy and Autophagy-Related Pathways in Cancer. Nature Reviews. Molecular Cell Biology 2023, 24: 560-575.

[44] Yu Z, Guo J, Hu M, Gao Y, Huang L: Icaritin Exacerbates Mitophagy and Synergizes with Doxorubicin to Induce Immunogenic Cell Death in Hepatocellular Carcinoma. ACS Nano 2020, 14: 4816-4828.

[45] Luo P, An Y, He J, Xing X, Zhang Q, Liu X, et al.: Icaritin with Autophagy/Mitophagy Inhibitors Synergistically Enhances Anticancer Efficacy and Apoptotic Effects Through PINK1/Parkin-Mediated Mitophagy in Hepatocellular Carcinoma. Cancer Letters 2024, 587: 216621.

[46] Li Z, Meng X, Jin L: Icaritin Induces Apoptotic and Autophagic Cell Death in Human Glioblastoma Cells. American Journal of Translational Research 2016, 8: 4628-4643.

[47] Pan XW, Li L, Huang Y, Huang H, Xu DF, Gao Y, et al.: Icaritin Acts Synergistically with Epirubicin to Suppress Bladder Cancer Growth Through Inhibition of Autophagy. Oncology Reports 2016, 35: 334-342.

[48] Zhou C, Gu J, Zhang G, Dong D, Yang Q, Chen MB, et al.: AMPK-Autophagy Inhibition Sensitizes Icaritin-Induced Anti-Colorectal Cancer Cell Activity. Oncotarget 2017, 8: 14736-14747.

[49] Zhang J, Xiang Q, Wu M, Lao YZ, Xian YF, Xu HX, et al.: Autophagy Regulators in Cancer. International Journal of Molecular Sciences 2023, 24: 10944.

[50] Su Z, Yang Z, Xu Y, Chen Y, Yu Q: Apoptosis, Autophagy, Necroptosis, and Cancer Metastasis. Molecular Cancer 2015, 14: 48.

[51] Vaseva AV, Marchenko ND, Ji K, Tsirka SE, Holzmann S, Moll UM: p53 Opens the Mitochondrial Permeability Transition Pore to Trigger Necrosis. Cell 2012, 149: 1536-1548.

[52] Zhou C, Chen Z, Lu X, Wu H, Yang Q, Xu D: Icaritin Activates JNK-Dependent mPTP Necrosis Pathway in Colorectal Cancer Cells. Tumour Biology: The Journal of the International Society for Oncodevelopmental Biology and Medicine 2016, 37: 3135-3144.

[53] Wang L, Lankhorst L: Exploiting Senescence for the Treatment of Cancer. Nature Review Cancer 2022, 22: 340-355.

[54] Zhu S, Xing C, Zhang G, Peng H, Wang Z: Icaritin Induces Cellular Senescence by Accumulating the ROS Production and Regulation of the Jak2/Stat3/p21 Pathway in Imatinib-Resistant, Chronic Myeloid Leukemia Cells. American Journal of Translational Research 2021, 13: 8860-8872.

[55] Wang S, Wang Q, Wang H, Qin C, Cui X, Li L, et al.: Induction of ROS and DNA Damage-Dependent Senescence by Icaritin Contributes to its Antitumor Activity in Hepatocellular Carcinoma Cells. Pharmaceutical Biology 2019, 57: 424-431.

[56] Jamasbi E, Hamelian M, Hossain MA, Varmira K: The Cell Cycle, Cancer Development and Therapy. Molecular Biology Reports 2022, 49: 10875-10883.

[57] Matthews HK, Bertoli C, de Bruin RAM: Cell Cycle Control in Cancer. Nature Review. Molecular Cell Biology 2022, 23: 74-88.

[58] Hong J, Zhang Z, Lv W, Zhang M, Chen C, Yang S, et al.: Icaritin Synergistically Enhances the Radiosensitivity of 4T1 Breast Cancer Cells. PLoS One 2013, 8: e71347.

[59] Zhu YH, Zhang XR, Zhang Q, Chai J: Icaritin-Elevated circ_0000190 Suppresses the Malignant Progression of Multiple Myeloma by Targeting miR-301a. Kaohsiung Journal of Medical Sciences 2022, 38: 447-456.

[60] Li C, Peng W, Song X, Wang Q, Wang W: Anticancer Effect of Icaritin Inhibits Cell Growth of Colon Cancer Through Reactive Oxygen Species, Bcl-2 and Cyclin D1/E Signaling. Oncology Letters 2016, 12: 3537-3542.

[61] Xiao Y, Yao W, Lin M, Huang W, Li B, Peng B, et al.: Icaritin-Loaded PLGA Nanoparticles Activate Immunogenic Cell Death and Facilitate Tumor Recruitment in Mice with Gastric Cancer. Drug Delivery 2022, 29: 1712-1725.

[62] Zhang C, Wang X, Zhang C: Icaritin Inhibits CDK2 Expression and Activity to Interfere with Tumor Progression. iScience 2022, 25: 104991.

[63] Huang X, Zhu D, Lou Y: A Novel Anticancer Agent, Icaritin, Induced Cell Growth Inhibition, G1 Arrest and Mitochondrial Transmembrane Potential Drop in Human Prostate Carcinoma PC-3 Cells. European Journal of Pharmacology 2007, 564: 26-36.

[64] Guo Y, Zhang X, Meng J, Wang ZY: An Anticancer Agent Icaritin Induces Sustained Activation of the Extracellular Signal-Regulated Kinase (ERK) Pathway and Inhibits Growth of Breast Cancer Cells. European Journal of Pharmacology 2011, 658: 114-122.

[65] Siegel RL, Miller KD, Jemal A: Cancer Statistics, 2020. CA: A Cancer Journal for Clinicians 2020, 70: 7-30.

[66] Ganesh K, Massague J: Targeting Metastatic Cancer. Nature Medicine 2021, 27: 34-44.

[67] Stoletov K, Beatty PH, Lewis JD: Novel Therapeutic Targets for Cancer Metastasis. Expert Review of Anticancer Therapy 2020, 20: 97-109.

[68] Li X, Zhang W, Liang L, Duan X, Deng J, Zhou Y: Natural Product-Derived Icaritin Exerts Anti-Glioblastoma Effects by Positively Modulating Estrogen Receptor β. Experimental and Therapeutic Medicine 2020, 19: 2841-2850.

[69] Pan C, Fujiwara Y, Horlad H, Shiraishi D, Iriki T, Tsuboki J, et al.: Flavonoid Compounds Contained in Epimedii Herba Inhibit Tumor Progression by Suppressing STAT3 Activation in the Tumor Microenvironment. Frontiers in Pharmacology 2020, 11: 262.

[70] Xu B, Jiang C, Han H, Liu H, Tang M, Liu L, et al.: Icaritin Inhibits the Invasion and Epithelial-to-Mesenchymal Transition of Glioblastoma Cells by Targeting EMMPRIN via PTEN/AKt/HIF-1alpha Signalling. Clinical and Experimental Pharmacology & Physiology 2015, 42: 1296-1307.

[71] Gao L, Ouyang Y, Li R, Zhang X, Gao X, Lin S, et al.: Icaritin Inhibits Migration and Invasion of Human Ovarian Cancer Cells via the Akt/mTOR Signaling Pathway. Frontiers in Oncology 2022, 12: 843489.

[72] Ramos RI, Bustos MA, Wu J, Jones P, Chang SC, Kiyohara E, et al.: Upregulation of Cell Surface GD3 Ganglioside Phenotype is Associated with Human Melanoma Brain Metastasis. Molecular Oncology 2020, 14: 1760-1778.

[73] Han S, Gou Y, Jin D, Ma J, Chen M, Dong X: Effects of Icaritin on the Physiological Activities of Esophageal Cancer Stem Cells. Biochemical and Biophysical Research Communications 2018, 504: 792-796.

[74] Tu S, Mao D, Shi M, Zhang H, Liu C, Li X, et al.: Icaritin Ameliorates Extracellular Microparticles-Induced Inflammatory Pre-Metastatic Niche via Modulating the cGAS-STING Signaling. Phytotherapy Research 2022, 36: 2127-2142.

[75] Guo Z, Jing X, Sun X, Sun S, Yang Y, Cao Y: Tumor Angiogenesis and Anti-Angiogenic Therapy. Chinese Medical Journal (Engl) 2024, 137: 2043-2051.

[76] Pathak A, Pal AK: Role of Angiogenesis and its Biomarkers in Development of Targeted Tumor Therapies. Stem Cells International 2024, 2024: 9077926.

[77] Yu X, Tong Y, Han XQ, Kwok HF, Yue GG, Lau CB, et al.: Anti-Angiogenic Activity of Herba Epimedii on Zebrafish Embryos In Vivo and HUVECs In Vitro. Phytotherapy Research 2013, 27: 1368-1375.

[78] Shi P, Hong J, Huang Y, Zhang Z, Zhang M, Zhang L: Automated Computational Framework of Blood Vessel Quantification in Chick Chorioallantoic Membrane Angiogenesis. Journal of Biomedical Optics 2014, 19: 106005.

[79] Guo J, Zeng H, Liu Y, Shi X, Liu Y, Liu C, et al.: Multicomponent Thermosensitive Lipid Complexes Enhance Desmoplastic Tumor Therapy Through Boosting Anti-Angiogenesis and Synergistic Strategy. International Journal of Pharmaceutics 2021, 601: 120533.

[80] Biserova K, Jakovlevs A: Cancer Stem Cells: Significance in Origin, Pathogenesis and Treatment of Glioblastoma. Cells 2021, 10: 621.

[81] Oskarsson T, Batlle E, Massague J: Metastatic Stem Cells: Sources, Niches, and Vital Pathways. Cell Stem Cell 2014, 14: 306-321.

[82] Zhao H, Guo Y, Li S, Han R, Ying J, Zhu H, et al.: A Novel Anti-Cancer Agent Icaritin Suppresses Hepatocellular Carcinoma Initiation and Malignant Growth Through the IL-6/Jak2/Stat3 Pathway. Oncotarget 2015, 6: 31927-31943.

[83] Chen M, Turhan AG, Ding H, Lin Q, Meng K, Jiang X: Targeting BCR-ABL+ Stem/Progenitor Cells and BCR-ABL-T315I Mutant Cells by Effective Inhibition of the BCR-ABL-Tyr177-GRB2 Complex. Oncotarget 2017, 8: 43662-43677.

[84] Salvadori G, Mirisola MG: Intermittent and Periodic Fasting, Hormones, and Cancer Prevention. Cancers (Basel) 2021, 13: 4587.

[85] Liang J, Shang Y: Estrogen and Cancer. Annual Review Physiology 2013, 75: 225-240.

[86] Wang ZQ, Lou YJ: Proliferation-Stimulating Effects of Icaritin and Desmethylicaritin in MCF-7 Cells. European Journal of Pharmacology 2004, 504: 147-153.

[87] Tiong CT, Chen C, Zhang SJ, Li J, Soshilov A, Denison MS, et al.: A Novel Prenylflavone Restricts Breast Cancer Cell Growth Through AhR-Mediated Destabilization of ERα Protein. Carcinogenesis 2012, 33: 1089-1097.

[88] Ma HR, Wang J, Chen YF, Chen H, Wang WS, Aisa HA: Icariin and Icaritin Stimulate the Proliferation of SKBr3 Cells Through the GPER1-Mediated Modulation of the EGFR-MAPK Signaling Pathway. International Journal of Molecular Medicien 2014, 33: 1627-1634.

[89] Yin L, Qi XW, Liu XZ, Yang ZY, Cai RL, Cui HJ, et al.: Icaritin Enhances the Efficacy of Cetuximab Against Triple-Negative Breast Cancer Cells. Oncology Letters 2020, 19: 3950-3958.

[90] Sun F, Zhang ZW, Tan EM, Lim ZLR, Li Y, Wang XC, et al.: Icaritin Suppresses Development of Neuroendocrine Differentiation of Prostate Cancer Through Inhibition of IL-6/STAT3 and Aurora Kinase A Pathways in TRAMP Mice. Carcinogenesis 2016, 37: 701-711.

[91] Sun F, Indran IR, Zhang ZW, Tan MH, Li Y, Lim ZL, et al.: A Novel Prostate Cancer Therapeutic Strategy Using Icaritin-Activated Arylhydrocarbon-Receptor to Co-Target Androgen Receptor and its Splice Variants. Carcinogenesis 2015, 36: 757-768.

[92] Jafari R, Almqvist H, Axelsson H, Ignatushchenko M, Lundbäck T, Nordlund P, et al.: The Cellular Thermal Shift Assay for Evaluating Drug Target Interactions in Cells. Nature Protocols 2014, 9: 2100-2122.

[93] Aurilio G, Cimadamore A, Mazzucchelli R, Lopez-Beltran A, Verri E, Scarpelli M, et al.: Androgen Receptor Signaling Pathway in Prostate Cancer: From Genetics to Clinical Applications. Cells 2020, 9: 2653.

[94] Sahoo BM, Banik BK, Borah P, Jain A: Reactive Oxygen Species (ROS): Key Components in Cancer Therapies. Anticancer Agents in Medicine Chemistry 2022, 22: 215-222.

[95] Assi M: The Differential Role of Reactive Oxygen Species in Early and Late Stages of Cancer. American Journal of Physiology. Regulatory, Integrative and Comparative Physiology 2017, 313: R646-R653.

[96] Zhang J, Wang X, Vikash V, Ye Q, Wu D, Liu Y, et al.: ROS and ROS-Mediated Cellular Signaling. Oxidative Medicine and Cellular Longevity 2016, 2016: 4350965.

[97] Zou J, Xu MX, Li F, Wang YH, Li XQ, Yu DJ, et al.: Icaritin Alleviates Docetaxel-Induced Skin Injury by Suppressing Reactive Oxygen Species via Estrogen Receptors. Thoracic Cancer 2022, 13: 190-201.

[98] Gorrini C, Harris IS, Mak TW: Modulation of Oxidative Stress as an Anticancer Strategy. Nature Reviews. Drug Discovery 2013, 12: 931-947.

[99] Shin MH, Oh E, Kim Y, Nam DH, Jeon SY, Yu JH, et al.: Recent Advances in CAR-Based Solid Tumor Immunotherapy. Cells 2023, 12: 1606.

[100] DeNardo DG, Ruffell B: Macrophages as Regulators of Tumour Immunity and Immunotherapy. Nature Reviews. Immunology 2019, 19: 369-382.

[101] Mantovani A, Marchesi F, Malesci A, Laghi L, Allavena P: Tumour-Associated Macrophages as Treatment Targets in Oncology. Nature Reviews Clinicals Oncology 2017, 14: 399-416.

[102] Mo D, Zhu H, Wang J, Hao H, Guo Y, Wang J, et al.: Icaritin Inhibits PD-L1 Expression by Targeting Protein IκB Kinase α. European Journal of Immunology 2021, 51: 978-988.

[103] Tao H, Liu M, Wang Y, Luo S, Xu Y, Ye B, et al.: Icaritin Induces Anti-tumor Immune Responses in Hepatocellular Carcinoma by Inhibiting Splenic Myeloid-Derived Suppressor Cell Generation. Frontiers in Immunology 2021, 12: 609295.

[104] Zhou J, Wu J, Chen X, Fortenbery N, Eksioglu E, Kodumudi KN, et al.: Icariin and its Derivative, ICT, Exert Anti-Inflammatory, Anti-Tumor Effects, and Modulate Myeloid Derived Suppressive Cells (MDSCs) Functions. International Immunopharmacology 2011, 11: 890-898.

[105] Binnewies M, Roberts EW, Kersten K, Chan V, Fearon DF, Merad M, et al.: Understanding the Tumor Immune Microenvironment (TIME) for Effective Therapy. Nature Medicine 2018, 24: 541-550.

[106] Hao H, Zhang Q, Zhu H, Wen Y, Qiu D, Xiong J, et al.: Icaritin Promotes Tumor T-cell Infiltration and Induces Antitumor Immunity in Mice. European Journal of Immunology 2019, 49: 2235-2244.

[107] Qin SK, Li Q, Ming Xu J, Liang J, Cheng Y, Fan Y, et al.: Icaritin-Induced Immunomodulatory Efficacy in Advanced Hepatitis B Virus-Related Hepatocellular Carcinoma: Immunodynamic Biomarkers and Overall Survival. Cancer Science 2020, 111: 4218-4231.

[108] Dongye Z, Wu X, Wen Y, Ding X, Wang C, Zhao T, et al.: Icaritin and Intratumoral Injection of CpG Treatment Synergistically Promote T Cell Infiltration and Antitumor Immune Response in Mice. International Immunopharmacology 2022, 111: 109093.

[109] Fan Y, Li S, Ding X, Yue J, Jiang J, Zhao H, et al.: First-In-Class Immune-Modulating Small Molecule Icaritin in Advanced Hepatocellular Carcinoma: Preliminary Results of Safety, Durable Survival and Immune biomarkers. BMC Cancer 2019, 19: 279.

[110] Sun Y, Qin S, Li W, Guo Y, Zhang Y, Meng L, et al.: A Randomized, Double-Blinded, Phase III Study of Icaritin Versus Huachashu as the First-Line Therapy in Biomarker-Enriched HBV-Related Advanced Hepatocellular Carcinoma with Poor Conditions: Interim Analysis Result. Journal of Clinical Oncology 2021, 39(15 suppl): abstract 4077.