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BIOLOGY MOLECULAR / CLINICAL RESEARCH
APOE suppresses osteosarcoma by modulating ferroptosis through the mTOR/Stat3 signaling pathway
1
Department of Orthopedics, Minhang Hospital, Fudan University, Minhang District, Shanghai, China
These authors had equal contribution to this work
Submission date: 2024-09-27
Final revision date: 2025-03-06
Acceptance date: 2025-04-13
Online publication date: 2025-04-20
Corresponding author
Liang Wu
Department of Orthopedics Minhang Hospital Fudan University No. 170 Xinsong Road Minhang District Shanghai, 201199, China
Department of Orthopedics Minhang Hospital Fudan University No. 170 Xinsong Road Minhang District Shanghai, 201199, China
KEYWORDS
TOPICS
ABSTRACT
Introduction:
Osteosarcoma (OS) is a highly malignant bone tumor with limited treatment options. The role of apolipoprotein E (APOE) in OS remains unclear. This study explores the impact of APOE overexpression on OS, particularly its effects on ferroptosis and autophagy.
Material and methods:
APOE was identified as a key gene through weighted gene co-expression network analysis (WGCNA) and protein-protein interaction (PPI) network analysis of the GSE28424 dataset. APOE was overexpressed in OS cell lines to evaluate its effects on cell behavior. The role of autophagy was investigated using the autophagy inhibitor 3-methyladenine (3-MA). The involvement of ferroptosis and the mTOR/Stat3 signaling pathway was investigated utilizing quantitative real-time reverse transcription PCR (qRT-PCR), Western blot (WB), and flow cytometry. A mouse xenograft model was employed to validate the in vitro results.
Results:
APOE overexpression significantly inhibited OS cell proliferation, invasion, migration, and epithelial-mesenchymal transition (EMT), with 3-MA partially reversing these effects. APOE overexpression also inhibited the mTOR and Stat3 expression, enhancing autophagy, as shown by increased LC3B-1, LC3B-2, and Beclin1 expression. Additionally, APOE overexpression promoted apoptosis, associated with increased reactive oxygen species (ROS) and intracellular Fe²+ levels, and altered ferroptosis-related gene expression, including upregulation of TfR1 and downregulation of FPN, GPX4, and SLC7A11. In vivo, APOE overexpression in a mouse xenograft model resulted in significantly smaller tumors, with changes in autophagy and ferroptosis markers consistent with in vitro findings.
Conclusions:
APOE overexpression suppresses osteosarcoma growth by promoting ferroptosis and autophagy through the mTOR/Stat3 signaling pathway, highlighting its promise as a target for OS therapeutic intervention.
Osteosarcoma (OS) is a highly malignant bone tumor with limited treatment options. The role of apolipoprotein E (APOE) in OS remains unclear. This study explores the impact of APOE overexpression on OS, particularly its effects on ferroptosis and autophagy.
Material and methods:
APOE was identified as a key gene through weighted gene co-expression network analysis (WGCNA) and protein-protein interaction (PPI) network analysis of the GSE28424 dataset. APOE was overexpressed in OS cell lines to evaluate its effects on cell behavior. The role of autophagy was investigated using the autophagy inhibitor 3-methyladenine (3-MA). The involvement of ferroptosis and the mTOR/Stat3 signaling pathway was investigated utilizing quantitative real-time reverse transcription PCR (qRT-PCR), Western blot (WB), and flow cytometry. A mouse xenograft model was employed to validate the in vitro results.
Results:
APOE overexpression significantly inhibited OS cell proliferation, invasion, migration, and epithelial-mesenchymal transition (EMT), with 3-MA partially reversing these effects. APOE overexpression also inhibited the mTOR and Stat3 expression, enhancing autophagy, as shown by increased LC3B-1, LC3B-2, and Beclin1 expression. Additionally, APOE overexpression promoted apoptosis, associated with increased reactive oxygen species (ROS) and intracellular Fe²+ levels, and altered ferroptosis-related gene expression, including upregulation of TfR1 and downregulation of FPN, GPX4, and SLC7A11. In vivo, APOE overexpression in a mouse xenograft model resulted in significantly smaller tumors, with changes in autophagy and ferroptosis markers consistent with in vitro findings.
Conclusions:
APOE overexpression suppresses osteosarcoma growth by promoting ferroptosis and autophagy through the mTOR/Stat3 signaling pathway, highlighting its promise as a target for OS therapeutic intervention.
REFERENCES (55)
1.
Czarnecka AM, Synoradzki K, Firlej W, et al. Molecular biology of osteosarcoma. Cancers 2020; 12: 2130.
2.
Beird HC, Bielack SS, Flanagan AM, et al. Osteosarcoma. Nat Rev Dis Primers 2022; 8: 77.
3.
Zarghooni K, Bratke G, Landgraf P, Simon T, Maintz D, Eysel P. The diagnosis and treatment of osteosarcoma and Ewing’s sarcoma in children and adolescents. Deutsches Ärzteblatt Int 2023; 120: 405.
4.
Tsukamoto S, Errani C, Angelini A, Mavrogenis AF. Current treatment considerations for osteosarcoma metastatic at presentation. Orthopedics 2020; 43: e345-58.
5.
Vitto VAM, Bianchin S, Zolondick AA, et al. Molecular mechanisms of autophagy in cancer development, progression, and therapy. Biomedicines 2022; 10: 1596.
6.
Liu K, Ren T, Huang Y, et al. Apatinib promotes autophagy and apoptosis through VEGFR2/STAT3/BCL-2 signaling in osteosarcoma. Cell Death Dis 2017; 8: e3015-e.
7.
Pu Y, Wang J, Wang S. Role of autophagy in drug resistance and regulation of osteosarcoma. Mol Clin Oncol 2022; 16: 72.
8.
Wang Yt, Tang F, Hu X, et al. Role of crosstalk between STAT3 and mTOR signaling in driving sensitivity to chemotherapy in osteosarcoma cell lines. IUBMB Life 2020; 72: 2146-53.
9.
Marafie SK, Al-Mulla F, Abubaker J. mTOR: its critical role in metabolic diseases, cancer, and the aging process. Int J Mol Sci 2024; 25: 6141.
10.
Song H, Liu J, Wu X, et al. LHX2 promotes malignancy and inhibits autophagy via mTOR in osteosarcoma and is negatively regulated by miR-129-5p. Aging (Albany NY) 2019; 11: 9794.
11.
Hu Y, Dong Z, Liu K. Unraveling the complexity of STAT3 in cancer: molecular understanding and drug discovery. J Exp Clin Cancer Res 2024; 43: 23.
12.
Liu Y, Liao S, Bennett S, et al. STAT3 and its targeting inhibitors in osteosarcoma. Cell Prolif 2021; 54: e12974.
13.
Ji Z, Shen J, Lan Y, Yi Q, Liu H. Targeting signaling pathways in osteosarcoma: mechanisms and clinical studies. MedComm 2023; 4: e308.
14.
Zhou Q, Meng Y, Li D, et al. Ferroptosis in cancer: from molecular mechanisms to therapeutic strategies. Signal Transduct Target Ther 2024; 9: 55.
15.
Consoli V, Fallica AN, Sorrenti V, Pittala V, Vanella L. Novel insights on ferroptosis modulation as potential strategy for cancer treatment: when nature kills. Antioxid Redox Signal 2024; 40: 40-85.
16.
Chen F, Cai X, Kang R, Liu J, Tang D. Autophagy-dependent ferroptosis in cancer. Antioxid Redox Signal 2023; 39: 79-101.
17.
Luo Y, Gao X, Zou L, Lei M, Feng J, Hu Z. Bavachin induces ferroptosis through the STAT3/P53/SLC7A11 axis in osteosarcoma cells. Oxid Med Cell Longev 2021; 2021: 1783485.
18.
Serrano-Pozo A, Das S, Hyman BT. APOE and Alzheimer’s disease: advances in genetics, pathophysiology, and therapeutic approaches. Lancet Neurol 2021; 20: 68-80.
19.
Zhao J, Gomes D, Yuan F, Feng J, Zhang X, O’Toole TE. Oral polystyrene consumption potentiates atherosclerotic lesion formation in ApoE(-/-) Mice. Circ Res 2024; 134: 1228-30.
20.
Alagarsamy J, Jaeschke A, Hui DY. Apolipoprotein E in cardiometabolic and neurological health and diseases. Int J Mol Sci 2022; 23: 9892.
21.
Wu L, Xian X, Tan Z, et al. The role of iron metabolism, lipid metabolism, and redox homeostasis in Alzheimer’s disease: from the perspective of ferroptosis. Mol Neurobiol 2023; 60: 2832-50.
22.
Peng X, Cai Z, Chen D, Ye F, Hong L. Prognostic significance and immune characteristics of APOE in gastric cancer. Aging (Albany NY) 2023; 15: 13840.
23.
Belaidi AA, Masaldan S, Southon A, et al. Apolipoprotein E potently inhibits ferroptosis by blocking ferritinophagy. Mol Psychiatry 2024; 29: 211-20.
24.
Argenziano M, Tortora C, Pota E, et al. Osteosarcoma in children: not only chemotherapy. Pharmaceuticals 2021; 14: 923.
25.
Synoradzki KJ, Bartnik E, Czarnecka AM, et al. TP53 in biology and treatment of osteosarcoma. Cancers 2021; 13: 4284.
26.
De Noon S, Ijaz J, Coorens TH, et al. MYC amplifications are common events in childhood osteosarcoma. J Pathol Clin Res 2021; 7: 425-31.
27.
Zhang Z, Liu B, Lin Z, Mei L, Chen R, Li Z. SPP1 could be an immunological and prognostic biomarker: from pan-cancer comprehensive analysis to osteosarcoma validation. FASEB J 2024; 38: e23783.
28.
Wang H, Han J, Dmitrii G, Ning K, Zhang X. KLF transcription factors in bone diseases. J Cell Mol Med 2024; 28: e18278.
29.
Lin H, Chen X, Zhang C, et al. EF24 induces ferroptosis in osteosarcoma cells through HMOX1. Biomed Pharmacother 2021; 136: 111202.
30.
Wu C, Li T, Cheng W. The correlation between APOE expression and the clinical characteristics and prognosis of patients with endometrial cancer. Medicine 2022; 101: e30536.
31.
Gan C, Zhang Y, Liang F, Guo X, Zhong Z. Effects of APOE gene ε4 allele on serum lipid profiles and risk of cardiovascular disease and tumorigenesis in southern Chinese population. World J Surg Oncol 2022; 20: 280.
32.
Zhou Y, Luo G. Apolipoproteins, as the carrier proteins for lipids, are involved in the development of breast cancer. Clin Transl Oncol 2020; 22: 1952-62.
33.
Liu S, Yao S, Yang H, Liu S, Wang Y. Autophagy: regulator of cell death. Cell Death Dis 2023; 14: 648.
34.
Ashrafizadeh M, Ahmadi Z, Farkhondeh T, Samarghandian S. Autophagy regulation using luteolin: new insight into its anti-tumor activity. Cancer Cell Int 2020; 20: 537.
35.
Lu Y, Wang Q, Zhou Y, et al. Overexpression of p62 is associated with poor prognosis and aggressive phenotypes in osteosarcoma. Oncol Letters 2018; 15: 9889-95.
36.
Liao YX, Lv JY, Zhou ZF, et al. CXCR4 blockade sensitizes osteosarcoma to doxorubicin by inducing autophagic cell death via PI3K Akt mTOR pathway inhibition. Int J Oncol 2021; 59: 49.
37.
Chang Z, Huo L, Li K, Wu Y, Hu Z. Blocked autophagy by miR-101 enhances osteosarcoma cell chemosensitivity in vitro. Sci World J 2014; 2014: 794756.
38.
Zou Z, Tao T, Li H, Zhu X. mTOR signaling pathway and mTOR inhibitors in cancer: progress and challenges. Cell Biosci 2020; 10: 31.
39.
Chen X, Cui Y, Ma Y. Long non-coding RNA BLACAT1 expedites osteosarcoma cell proliferation, migration and invasion via up-regulating SOX12 through miR-608. J Bone Oncol 2020; 25: 100314.
40.
Malla R, Marni R, Chakraborty A. Exploring the role of CD151 in the tumor immune microenvironment: therapeutic and clinical perspectives. Biochim Biophys Acta Rev Cancer 2023; 1878: 188898.
41.
Yang J, Wang L, Guan X, Qin JJ. Inhibiting STAT3 signaling pathway by natural products for cancer prevention and therapy: in vitro and in vivo activity and mechanisms of action. Pharmacol Res 2022; 182: 106357.
42.
Zhang W, Gong M, Zhang W, et al. Thiostrepton induces ferroptosis in pancreatic cancer cells through STAT3/GPX4 signalling. Cell Death Dis 2022; 13: 630.
43.
Li S, Huang Y. Ferroptosis: an iron-dependent cell death form linking metabolism, diseases, immune cell and targeted therapy. Clin Transl Oncol 2022; 24: 1-12.
44.
Li X, Liu J. FANCD2 inhibits ferroptosis by regulating the JAK2/STAT3 pathway in osteosarcoma. BMC Cancer 2023; 23: 179.
45.
Lv H, Zhen C, Liu J, Shang P. PEITC triggers multiple forms of cell death by GSH-iron-ROS regulation in K7M2 murine osteosarcoma cells. Acta Pharmacol Sin 2020; 41: 1119-32.
46.
Wen R, Dong X, Zhuang H, et al. Baicalin induces ferroptosis in osteosarcomas through a novel Nrf2/xCT/GPX4 regulatory axis. Phytomedicine 2023; 116: 154881.
47.
Stockwell BR, Friedmann Angeli JP, Bayir H, et al. Ferroptosis: a regulated cell death nexus linking metabolism, redox biology, and disease. Cell 2017; 171: 273-85.
48.
Marais AD. Apolipoprotein E in lipoprotein metabolism, health and cardiovascular disease. Pathology 2019; 51: 165-76.
49.
Huang Y, Liu XQ, Rall SC Jr, et al. Overexpression and accumulation of apolipoprotein E as a cause of hypertriglyceridemia. J Biol Chem 1998; 273: 26388-93.
50.
Ma HQ, Cui LH, Li CC, Yu Z, Piao JM. Effects of serum triglycerides on prostate cancer and breast cancer risk: a meta-analysis of prospective studies. Nutr Cancer 2016; 68: 1073-82.
51.
Wan Y, Zhang J, Chen M, Ma M, Sheng B. Elevated serum triglyceride levels may be a key independent predicting factor for gallbladder cancer risk in gallbladder stone disease patients: a case-control study. BMC Endocr Disord 2022; 22: 270.
52.
Broadfield LA, Pane AA, Talebi A, Swinnen JV, Fendt SM. Lipid metabolism in cancer: new perspectives and emerging mechanisms. Develop Cell 2021; 56: 1363-93.
53.
Kemp SB, Carpenter ES, Steele NG, et al. Apolipoprotein E promotes immune suppression in pancreatic cancer through NF-B-mediated production of CXCL1. Cancer Res 2021; 81: 4305-18.
54.
Sohn HY, Kim SI, Park JY, et al. ApoE4 attenuates autophagy via FoxO3a repression in the brain. Sci Rep 2021; 11: 17604.
55.
Parcon PA, Balasubramaniam M, Ayyadevara S, et al. Apolipoprotein E4 inhibits autophagy gene products through direct, specific binding to CLEAR motifs. Alzheimer’s Dementia 2018; 14: 230-42.
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