FOSL1 promotes stem cell‑like characteristics and anoikis resistance to facilitate tumorigenesis and metastasis in osteosarcoma by targeting SOX2

Wang,Yang; Hu,Qin; Cao,Ya; Yao,Li; Liu,Haoran; Wen,Yafeng; Bao,Yixi; Zhang,Shun; Lv,Chuanzhu; Zhao,Guo-Sheng

doi:10.3892/ijmm.2024.5418

FOSL1 promotes stem cell‑like characteristics and anoikis resistance to facilitate tumorigenesis and metastasis in osteosarcoma by targeting SOX2

Authors:
- Yang Wang
- Qin Hu
- Ya Cao
- Li Yao
- Haoran Liu
- Yafeng Wen
- Yixi Bao
- Shun Zhang
- Chuanzhu Lv
- Guo-Sheng Zhao
View Affiliations

Affiliations: Department of Emergency Medicine Center, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan 610064, P.R. China, Department of Emergency Medicine Center, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan 610064, P.R. China, Department of Pathology, Xinqiao Hospital, The Third Military Medical University, Chongqing 400037, P.R. China, Department of Spine Surgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, P.R. China, Department of Clinical Laboratory, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, P.R. China, Department of Spine Surgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, P.R. China
Published online on: August 26, 2024 https://doi.org/10.3892/ijmm.2024.5418
Article Number: 94
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Metastasis is the leading cause of cancer‑related death in osteosarcoma (OS). OS stem cells (OSCs) and anoikis resistance are considered to be essential for tumor metastasis formation. However, the underlying mechanisms involved in the maintenance of a stem‑cell phenotype and anoikis resistance in OS are mostly unknown. Fos‑like antigen 1 (FOSL1) is important in maintaining a stem‑like phenotype in various cancers; however, its role in OSCs and anoikis resistance remains unclear. In the present study, the dynamic expression patterns of FOSL1 were investigated during the acquisition of cancer stem‑like properties using RNA sequencing, PCR, western blotting and immunofluorescence. Flow cytometry, tumor‑sphere formation, clone formation assays, anoikis assays, western blotting and in vivo xenograft and metastasis models were used to further investigate the responses of the stem‑cell phenotype and anoikis resistance to FOSL1 overexpression or silencing in OS cell lines. The underlying molecular mechanisms were evaluated, focusing on whether SOX2 is crucially involved in FOSL1‑mediated stemness and anoikis in OS. FOSL1 expression was observed to be upregulated in OSCs and promoted tumor‑sphere formation, clone formation and tumorigenesis in OS cells. FOSL1 expression correlated positively with the expression of stemness‑related factors (SOX2, NANOG, CD117 and Stro1). Moreover, FOSL1 facilitated OS cell anoikis resistance and promoted metastases by regulating the expression of apoptosis related proteins BCL2 and BAX. Mechanistically, FOSL1 upregulated SOX2 expression by interacting with the SOX2 promoter and activating its transcription. The results also showed that SOX2 is critical for FOSL1‑mediated stem‑like properties and anoikis resistance. The current findings indicated that FOSL1 is an important regulator that promotes a stem cell‑like phenotype and anoikis resistance to facilitate tumorigenesis and metastasis in OS by regulating the transcription of SOX2. Thus, FOSL1 might represent an attractive target for therapeutic interventions in OS.

View Figures

View References

1	Siegel RL, Giaquinto AN and Jemal A: Cancer statistics, 2024. CA Cancer J Clin. 74:12–49. 2024. View Article : Google Scholar : PubMed/NCBI
2	Zheng R, Zhang S, Zeng H, Wang S, Sun K, Chen R, Li L, Wei W and He J: Cancer incidence and mortality in China, 2016. J Natl Cancer Cent. 2:1–9. 2022. View Article : Google Scholar : PubMed/NCBI
3	Ferguson JL and Turner SP: Bone Cancer: Diagnosis and treatment principles. Am Fam Physician. 98:205–213. 2018.PubMed/NCBI
4	Mensali N, Köksal H, Joaquina S, Wernhoff P, Casey NP, Romecin P, Panisello C, Rodriguez R, Vimeux L, Juzeniene A, et al: ALPL-1 is a target for chimeric antigen receptor therapy in osteosarcoma. Nat Commun. 14:33752023. View Article : Google Scholar : PubMed/NCBI
5	Zhao GS, Gao ZR, Zhang Q, Tang XF, Lv YF, Zhang ZS, Zhang Y, Tan QL, Peng DB, Jiang DM and Guo QN: TSSC3 promotes autophagy via inactivating the Src-mediated PI3K/Akt/mTOR pathway to suppress tumorigenesis and metastasis in osteosarcoma, and predicts a favorable prognosis. J Exp Clin Cancer Res. 37:1882018. View Article : Google Scholar : PubMed/NCBI
6	Kim YI, Tseng YC, Ayaz G, Wang S, Yan H, du Bois W, Yang H, Zhen T, Lee MP, Liu P, et al: SOX9 is a key component of RUNX2-regulated transcriptional circuitry in osteosarcoma. Cell Biosci. 13:1362023. View Article : Google Scholar : PubMed/NCBI
7	Zhang S, Zhu N, Li HF, Gu J, Zhang CJ, Liao DF and Qin L: The lipid rafts in cancer stem cell: A target to eradicate cancer. Stem Cell Res Ther. 13:4322022. View Article : Google Scholar : PubMed/NCBI
8	Gibbs CP, Kukekov VG, Reith JD, Tchigrinova O, Suslov ON, Scott EW, Ghivizzani SC, Ignatova TN and Steindler DA: Stem-like cells in bone sarcomas: Implications for tumorigenesis. Neoplasia. 7:967–976. 2005. View Article : Google Scholar : PubMed/NCBI
9	Yan GN, Lv YF and Guo QN: Advances in osteosarcoma stem cell research and opportunities for novel therapeutic targets. Cancer Lett. 370:268–274. 2016. View Article : Google Scholar
10	Martins-Neves SR, Sampaio-Ribeiro G and Gomes CMF: Self-Renewal and pluripotency in osteosarcoma stem cells' chemoresistance: Notch, Hedgehog, and Wnt/β-Catenin Interplay with Embryonic Markers. Int J Mol Sci. 24:84012023. View Article : Google Scholar
11	Ullmann P, Rodriguez F, Schmitz M, Meurer SK, Qureshi-Baig K, Felten P, Ginolhac A, Antunes L, Frasquilho S, Zügel N, et al: The miR-371~373 cluster represses colon cancer initiation and metastatic colonization by inhibiting the TGFBR2/ID1 signaling axis. Cancer Res. 78:3793–3808. 2018. View Article : Google Scholar : PubMed/NCBI
12	Taddei ML, Giannoni E, Fiaschi T and Chiarugi P: Anoikis: An emerging hallmark in health and diseases. J Pathol. 226:380–393. 2012. View Article : Google Scholar
13	Paoli P, Giannoni E and Chiarugi P: Anoikis molecular pathways and its role in cancer progression. Biochim Biophys Acta. 1833:3481–3498. 2013. View Article : Google Scholar : PubMed/NCBI
14	Han YH, Wang Y, Lee SJ, Jin MH, Sun HN and Kwon T: Regulation of anoikis by extrinsic death receptor pathways. Cell Commun Signal. 21:2272023. View Article : Google Scholar : PubMed/NCBI
15	Sattari Fard F, Jalilzadeh N, Mehdizadeh A, Sajjadian F and Velaei K: Understanding and targeting anoikis in metastasis for cancer therapies. Cell Biol Int. 47:683–698. 2023. View Article : Google Scholar
16	Sun T, Zhong X, Song H, Liu J, Li J, Leung F, Lu WW and Liu ZL: Anoikis resistant mediated by FASN promoted growth and metastasis of osteosarcoma. Cell Death Dis. 10:2982019. View Article : Google Scholar : PubMed/NCBI
17	Zhao GS, Zhang Q, Cao Y, Wang Y, Lv YF, Zhang ZS, Zhang Y, Tan QL, Chang Y, Quan ZX, et al: High expression of ID1 facilitates metastasis in human osteosarcoma by regulating the sensitivity of anoikis via PI3K/AKT depended suppression of the intrinsic apoptotic signaling pathway. Am J Transl Res. 11:2117–2139. 2019.PubMed/NCBI
18	Zhang M, Hoyle RG, Ma Z, Sun B, Cai W, Cai H, Xie N, Zhang Y, Hou J, Liu X, et al: FOSL1 promotes metastasis of head and neck squamous cell carcinoma through super-enhancer-driven transcription program. Mol Ther. 29:2583–2600. 2021. View Article : Google Scholar : PubMed/NCBI
19	Jiang X, Xie H, Dou Y, Yuan J, Zeng D and Xiao S: Expression and function of FRA1 protein in tumors. Mol Biol Rep. 47:737–752. 2020. View Article : Google Scholar
20	Guo S, Ramar V, Guo AA, Saafir T, Akpobiyeri H, Hudson B, Li J and Liu M: TRPM7 transactivates the FOSL1 gene through STAT3 and enhances glioma stemness. Cell Mol Life Sci. 80:2702023. View Article : Google Scholar : PubMed/NCBI
21	Wang T, Song P, Zhong T, Wang X, Xiang X, Liu Q, Chen H, Xia T, Liu H, Niu Y, et al: The inflammatory cytokine IL-6 induces FRA1 deacetylation promoting colorectal cancer stem-like properties. Oncogene. 38:4932–4947. 2019. View Article : Google Scholar : PubMed/NCBI
22	Zhang K, Myllymäki SM, Gao P, Devarajan R, Kytölä V, Nykter M, Wei GH and Manninen A: Oncogenic K-Ras upregulates ITGA6 expression via FOSL1 to induce anoikis resistance and synergizes with αV-Class integrins to promote EMT. Oncogene. 36:5681–5694. 2017. View Article : Google Scholar : PubMed/NCBI
23	Shen H, Wang W, Ni B, Zou Q, Lu H and Wang Z: Exploring the molecular mechanisms of osteosarcoma by the integrated analysis of mRNAs and miRNA microarrays. Int J Mol Med. 42:21–30. 2018.PubMed/NCBI
24	Chandrashekar DS, Karthikeyan SK, Korla PK, Patel H, Shovon AR, Athar M, Netto GJ, Qin ZS, Kumar S, Manne U, et al: UALCAN: An update to the integrated cancer data analysis platform. Neoplasia. 25:18–27. 2022. View Article : Google Scholar : PubMed/NCBI
25	Zhao SJ, Jiang YQ, Xu NW, Li Q, Zhang Q, Wang SY, Li J, Wang YH, Zhang YL, Jiang SH, et al: SPARCL1 suppresses osteosarcoma metastasis and recruits macrophages by activation of canonical WNT/β-catenin signaling through stabilization of the WNT-receptor complex. Oncogene. 37:1049–1061. 2018. View Article : Google Scholar
26	Bartha Á and Győrffy B: TNMplot.com: A web tool for the comparison of gene expression in normal, tumor and metastatic tissues. Int J Mol Sci. 22:26222021. View Article : Google Scholar : PubMed/NCBI
27	Győrffy B: Discovery and ranking of the most robust prognostic biomarkers in serous ovarian cancer. Geroscience. 45:1889–1898. 2023. View Article : Google Scholar
28	Yang Y, Huang H, Li L and Yang Y: Multiplex Immunohistochemistry Staining for Paraffin-embedded lung cancer tissue. J Vis Exp. 2023. View Article : Google Scholar
29	Wang Y, Zhang L, Zhao G, Zhang Y, Zhan F, Chen Z, He T, Cao Y, Hao L, Wang Z, et al: Homologous targeting nanoparticles for enhanced PDT against osteosarcoma HOS cells and the related molecular mechanisms. J Nanobiotechnology. 20:832022. View Article : Google Scholar : PubMed/NCBI
30	Yan GN, Tang XF, Zhang XC, He T, Huang YS, Zhang X, Meng G, Guo DY, Lv YF and Guo QN: TSSC3 represses self-renewal of osteosarcoma stem cells and Nanog expression by inhibiting the Src/Akt pathway. Oncotarget. 8:85628–85641. 2017. View Article : Google Scholar : PubMed/NCBI
31	Naito S, von Eschenbach AC, Giavazzi R and Fidler IJ: Growth and metastasis of tumor cells isolated from a human renal cell carcinoma implanted into different organs of nude mice. Cancer Res. 46:4109–4115. 1986.PubMed/NCBI
32	Hu Y and Smyth GK: ELDA: Extreme limiting dilution analysis for comparing depleted and enriched populations in stem cell and other assays. J Immunol Methods. 347:70–78. 2009. View Article : Google Scholar : PubMed/NCBI
33	Adhikari AS, Agarwal N, Wood BM, Porretta C, Ruiz B, Pochampally RR and Iwakuma T: CD117 and Stro-1 identify osteosarcoma tumor-initiating cells associated with metastasis and drug resistance. Cancer Res. 70:4602–4612. 2010. View Article : Google Scholar : PubMed/NCBI
34	Marques C, Unterkircher T, Kroon P, Oldrini B, Izzo A, Dramaretska Y, Ferrarese R, Kling E, Schnell O, Nelander S, et al: NF1 regulates mesenchymal glioblastoma plasticity and aggressiveness through the AP-1 transcription factor FOSL1. Elife. 10:e648462021. View Article : Google Scholar : PubMed/NCBI
35	Gambera S, Abarrategi A, Rodríguez-Milla MA, Mulero F, Menéndez ST, Rodriguez R, Navarro S and García-Castro J: Role of Activator Protein-1 complex on the phenotype of human osteosarcomas generated from mesenchymal stem cells. Stem Cells. 36:1487–1500. 2018. View Article : Google Scholar : PubMed/NCBI
36	Whittle SB, Offer K, Roberts RD, LeBlanc A, London C, Majzner RG, Huang AY, Houghton P, Alejandro Sweet Cordero E, Grohar PJ, et al: Charting a path for prioritization of novel agents for clinical trials in osteosarcoma: A report from the Children's Oncology Group New Agents for Osteosarcoma Task Force. Pediatr Blood Cancer. 68:e291882021. View Article : Google Scholar : PubMed/NCBI
37	Hyakusoku H, Sawakuma K, Sano D, Takahashi H, Hatano T, Sato K, Isono Y, Shimada S, Takada K, Kuwahara T, et al: FosL1 regulates regional metastasis of head and neck squamous cell carcinoma by promoting cell migration, invasion, and proliferation. Anticancer Res. 41:3317–3326. 2021. View Article : Google Scholar : PubMed/NCBI
38	Dai Y, Zhang X, Ou Y, Zou L, Zhang D, Yang Q, Qin Y, Du X, Li W, Yuan Z, et al: Anoikis resistance-protagonists of breast cancer cells survive and metastasize after ECM detachment. Cell Commun Signal. 21:1902023. View Article : Google Scholar
39	Celià-Terrassa T and Kang Y: Distinctive properties of metastasis-initiating cells. Genes Dev. 30:892–908. 2016. View Article : Google Scholar : PubMed/NCBI
40	Lee BK, Uprety N, Jang YJ, Tucker SK, Rhee C, LeBlanc L, Beck S and Kim J: Fosl1 overexpression directly activates trophoblast-specific gene expression programs in embryonic stem cells. Stem Cell Res. 26:95–102. 2018. View Article : Google Scholar :
41	Qi XT, Li YL, Zhang YQ, Xu T, Lu B, Fang L, Gao JQ, Yu LS, Zhu DF, Yang B, et al: KLF4 functions as an oncogene in promoting cancer stem cell-like characteristics in osteosarcoma cells. Acta Pharmacol Sin. 40:546–555. 2019. View Article : Google Scholar :
42	Yang L, Shi P, Zhao G, Xu J, Peng W, Zhang J, Zhang G, Wang X, Dong Z, Chen F and Cui H: Targeting cancer stem cell pathways for cancer therapy. Signal Transduct Target Ther. 5:82020. View Article : Google Scholar : PubMed/NCBI
43	Shonibare Z, Monavarian M, O'Connell K, Altomare D, Shelton A, Mehta S, Jaskula-Sztul R, Phaeton R, Starr MD, Whitaker R, et al: Reciprocal SOX2 regulation by SMAD1-SMAD3 is critical for anoikis resistance and metastasis in cancer. Cell Rep. 40:1110662022. View Article : Google Scholar : PubMed/NCBI
44	Batlle E and Clevers H: Cancer stem cells revisited. Nat Med. 23:1124–1134. 2017. View Article : Google Scholar : PubMed/NCBI
45	Guha D, Saha T, Bose S, Chakraborty S, Dhar S, Khan P, Adhikary A, Das T and Sa G: Integrin-EGFR interaction regulates anoikis resistance in colon cancer cells. Apoptosis. 24:958–971. 2019. View Article : Google Scholar : PubMed/NCBI
46	Chen Z, Wang S, Li HL, Luo H, Wu X, Lu J, Wang HW, Chen Y, Chen D, Wu WT, et al: FOSL1 promotes proneural-to-mesenchymal transition of glioblastoma stem cells via UBC9/CYLD/NF-κB axis. Mol Ther. 30:2568–2583. 2022. View Article : Google Scholar : PubMed/NCBI
47	Li R, Che W, Liang N, Deng S, Song Z and Yang L: Silent FOSL1 enhances the radiosensitivity of glioma stem cells by down-regulating miR-27a-5p. Neurochem Res. 46:3222–3246. 2021. View Article : Google Scholar : PubMed/NCBI
48	Talukdar S, Pradhan AK, Bhoopathi P, Shen XN, August LA, Windle JJ, Sarkar D, Furnari FB, Cavenee WK, Das SK, et al: MDA-9/Syntenin regulates protective autophagy in anoikis-resistant glioma stem cells. Proc Natl Acad Sci USA. 115:5768–5773. 2018. View Article : Google Scholar : PubMed/NCBI
49	Kim SY, Hong SH, Basse PH, Wu C, Bartlett DL, Kwon YT and Lee YJ: Cancer stem cells protect non-stem cells from Anoikis: Bystander effects. J Cell Biochem. 117:2289–2301. 2016. View Article : Google Scholar : PubMed/NCBI
50	Xiong G, Ouyang S, Xie N, Xie J, Wang W, Yi C, Zhang M, Xu X, Chen D and Wang C: FOSL1 promotes tumor growth and invasion in ameloblastoma. Front Oncol. 12:9001082022. View Article : Google Scholar : PubMed/NCBI
51	Basu-Roy U, Seo E, Ramanathapuram L, Rapp TB, Perry JA, Orkin SH, Mansukhani A and Basilico C: Sox2 maintains self renewal of tumor-initiating cells in osteosarcomas. Oncogene. 31:2270–2282. 2012. View Article : Google Scholar
52	Ren C, Ren T, Yang K, Wang S, Bao X, Zhang F and Guo W: Inhibition of SOX2 induces cell apoptosis and G1/S arrest in Ewing's sarcoma through the PI3K/Akt pathway. J Exp Clin Cancer Res. 35:442016. View Article : Google Scholar : PubMed/NCBI