Hypoxia increases the expression of stem cell markers in human osteosarcoma cells

Lin,Jinluan; Wang,Xinwu; Wang,Xinwen; Wang,Shenglin; Shen,Rongkai; Yang,Yanbing; Xu,Jianyong; Lin,Jianhua

doi:10.3892/ol.2021.12478

Hypoxia increases the expression of stem cell markers in human osteosarcoma cells

Authors:
- Jinluan Lin
- Xinwu Wang
- Xinwen Wang
- Shenglin Wang
- Rongkai Shen
- Yanbing Yang
- Jianyong Xu
- Jianhua Lin
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Affiliations: Department of Orthopedics, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350005, P.R. China, Department of Orthopedics, The First Hospital of Putian City, Putian, Fujian 351100, P.R. China, Department of Orthopedics, The Third Affiliated Hospital of Southern Medical University, Guangzhou, Guangdong 510630, P.R. China, Department of Radiology, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, Fujian 350005, P.R. China, Department of Orthopedics, The People's Hospital of Guixi, Guixi, Jiangxi 335400, P.R. China
Published online on: January 20, 2021 https://doi.org/10.3892/ol.2021.12478
Article Number: 217
Copyright: © Lin et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Osteosarcoma (OS) is the most common primary malignant tumor of bone. It is a common phenomenon that osteosarcoma cells have a hypoxic microenvironment. Hypoxia can dedifferentiate cells of several malignant tumor types into stem cell‑like phenotypes. However, the role of hypoxia in stemness induction and the expression of cancer stem cell (CSC) markers in human osteosarcoma cells has not been reported. The present study examined the effects of hypoxia on stem‑like cells in the human osteosarcoma MNNG/HOS cells. Under the incubation with 1% oxygen, the expression of CSCs markers (Oct‑4, Nanog and CD133) in MNNG/HOS cells were increased. Moreover, MNNG/HOS cells cultured under hypoxic conditions were more likely to proliferate into spheres and resulted in larger xenograft tumor. Hypoxia also increased the mRNA and protein levels of hypoxia‑inducible factor (HIF)‑1α. Then rapamycin was used, which has been shown to lower HIF‑1α protein level, to inhibit the hypoxic response. Rapamycin suppressed the expression of HIF‑1α protein and CSCs markers (Oct4, Nanog and CD133) in MNNG/HOS cells. In addition, pretreatment with rapamycin reduced the efficiency of MNNG/HOS cells in forming spheres and xenograft tumors. The results demonstrated that hypoxia (1% oxygen) can dedifferentiate some of the MNNG/HOS cells into stem cell‑like phenotypes, and that the mTOR signaling pathway participates in this process via regulating the expression of HIF‑1α protein.

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1	Huang CY, Wei PL, Wang JW, Makondi PT, Huang MT, Chen HA and Chang YJ: Glucose-regulated protein 94 modulates the response of osteosarcoma to chemotherapy. Dis Markers. 2019:45697182019. View Article : Google Scholar : PubMed/NCBI
2	Urciuoli E, Giorda E, Scarsella M, Petrini S and Peruzzi B: Osteosarcoma-derived extracellular vesicles induce a tumor-like phenotype in normal recipient cells. J Cell Physiol. 233:6158–6172. 2018. View Article : Google Scholar : PubMed/NCBI
3	Li Y, Zeng C, Tu M, Jiang W, Dai Z, Hu Y, Deng Z and Xiao W: MicroRNA-200b acts as a tumor suppressor in osteosarcoma via targeting ZEB1. Onco Targets Ther. 9:3101–3111. 2016.PubMed/NCBI
4	Zhang Z, Luo G, Yu C, Yu G, Jiang R and Shi X: MicroRNA-493-5p inhibits proliferation and metastasis of osteosarcoma cells by targeting Kruppel-like factor 5. J Cell Physiol. 234:13525–13533. 2019. View Article : Google Scholar : PubMed/NCBI
5	Corrò C and Moch H: Biomarker discovery for renal cancer stem cells. J Pathol Clin Res. 4:3–18. 2018. View Article : Google Scholar : PubMed/NCBI
6	Wahab SMR, Islam F, Gopalan V and Lam AK: The identifications and clinical implications of cancer stem cells in colorectal cancer. Clin Colorectal Cancer. 16:93–102. 2017. View Article : Google Scholar : PubMed/NCBI
7	Yi M, Li J, Chen S, Cai J, Ban Y, Peng Q, Zhou Y, Zeng Z, Peng S, Li X, Xiong W, Li G and Xiang B: Correction to: Emerging role of lipid metabolism alterations in Cancer stem cells. J Exp Clin Cancer Res. 37:1552018. View Article : Google Scholar : PubMed/NCBI
8	Jiang P, Zhang Y, Zhu C, Zhang W, Mao Z and Gao C: Fe₃O₄/BSA particles induce osteogenic differentiation of mesenchymal stem cells under static magnetic field. Acta Biomater. 46:141–150. 2016. View Article : Google Scholar : PubMed/NCBI
9	Prasad P, Mittal SA, Chongtham J, Mohanty S and Srivastava T: Hypoxia-mediated epigenetic regulation of stemness in brain tumor cells. Stem Cells. 35:1468–1478. 2017. View Article : Google Scholar : PubMed/NCBI
10	Lin Q and Yun Z: Impact of the hypoxic tumor microenvironment on the regulation of cancer stem cell characteristics. Cancer Biol Ther. 9:949–956. 2010. View Article : Google Scholar : PubMed/NCBI
11	Couvelard A, O'Toole D, Turley H, Leek R, Sauvanet A, Degott C, Ruszniewski P, Belghiti J, Harris AL, Gatter K and Pezzella F: Microvascular density and hypoxia-inducible factor pathway in pancreatic endocrine tumours: negative correlation of microvascular density and VEGF expression with tumour progression. Br J Cancer. 92:94–101. 2005. View Article : Google Scholar : PubMed/NCBI
12	Li Z, Bao S, Wu Q, Wang H, Eyler C, Sathornsumetee S, Shi Q, Cao Y, Lathia J, McLendon RE, et al: Hypoxia-inducible factors regulate tumorigenic capacity of glioma stem cells. Cancer Cell. 15:501–513. 2009. View Article : Google Scholar : PubMed/NCBI
13	Kim Y, Lin Q, Glazer PM and Yun Z: Hypoxic tumor microenvironment and cancer cell differentiation. Curr Mol Med. 9:425–434. 2009. View Article : Google Scholar : PubMed/NCBI
14	Yuan G, Nanduri J, Khan S, Semenza GL and Prabhakar NR: Induction of HIF-1alpha expression by intermittent hypoxia: Involvement of NADPH oxidase, Ca2⁺ signaling, prolyl hydroxylases, and mTOR. J Cell Physiol. 217:674–685. 2008. View Article : Google Scholar : PubMed/NCBI
15	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
16	Lin X, Sun B, Zhu D, Zhao X, Sun R, Zhang Y, Zhang D, Dong X, Gu Q, Li Y, et al: Notch4⁺ cancer stem-like cells promote the metastatic and invasive ability of melanoma. Cancer Sci. 107:1079–1091. 2016. View Article : Google Scholar : PubMed/NCBI
17	Fan Z, Li M, Chen X, Wang J, Liang X, Wang H, Wang Z, Cheng B and Xia J: Prognostic value of cancer stem cell markers in head and neck squamous cell carcinoma: a meta-analysis. Sci Rep. 7:430082017. View Article : Google Scholar : PubMed/NCBI
18	Bradshaw A, Wickremsekera A, Tan ST, Peng L, Davis PF and Itinteang T: Cancer stem cell hierarchy in glioblastoma multiforme. Front Surg. 3:212016. View Article : Google Scholar : PubMed/NCBI
19	Wang L, Park P, Zhang H, La Marca F and Lin CY: Prospective identification of tumorigenic osteosarcoma cancer stem cells in OS99-1 cells based on high aldehyde dehydrogenase activity. Int J Cancer. 128:294–303. 2011. View Article : Google Scholar : PubMed/NCBI
20	Zhao Y, Zhao W, Lim YC and Liu T: Salinomycin-loaded gold nanoparticles for treating cancer stem cells by ferroptosis-induced cell death. Mol Pharm. 16:2532–2539. 2019. View Article : Google Scholar : PubMed/NCBI
21	Zeng L, Cen Y and Chen J: Long non-coding RNA MALAT-1 contributes to maintenance of stem cell-like phenotypes in breast cancer cells. Oncol Lett. 15:2117–2122. 2018.PubMed/NCBI
22	Mukherjee S, Baidoo JNE, Sampat S, Mancuso A, David L, Cohen LS, Zhou S and Banerjee P: Liposomal TriCurin, a synergistic combination of curcumin, epicatechin gallate and resveratrol, repolarizes tumor-associated microglia/macrophages, and eliminates glioblastoma (GBM) and GBM stem cells. Molecules. 23:2012018. View Article : Google Scholar
23	Krishnapriya S, Sidhanth C, Manasa P, Sneha S, Bindhya S, Nagare RP, Ramachandran B, Vishwanathan P, Murhekar K, Shirley S, et al: Cancer stem cells contribute to angiogenesis and lymphangiogenesis in serous adenocarcinoma of the ovary. Angiogenesis. 22:441–455. 2019. View Article : Google Scholar : PubMed/NCBI
24	Fourneaux B, Bourdon A, Dadone B, Lucchesi C, Daigle SR, Richard E, Laroche-Clary A, Le Loarer F and Italiano A: Identifying and targeting cancer stem cells in leiomyosarcoma: Prognostic impact and role to overcome secondary resistance to PI3K/mTOR inhibition. J Hematol Oncol. 12:112019. View Article : Google Scholar : PubMed/NCBI
25	Mohlin S, Wigerup C, Jögi A and Påhlman S: Hypoxia, pseudohypoxia and cellular differentiation. Exp Cell Res. 356:192–196. 2017. View Article : Google Scholar : PubMed/NCBI
26	Wang P, Lan C, Xiong S, Zhao X, Shan Y, Hu R, Wan W, Yu S, Liao B, Li G, et al: HIF1α regulates single differentiated glioma cell dedifferentiation to stem-like cell phenotypes with high tumorigenic potential under hypoxia. Oncotarget. 8:28074–28092. 2017. View Article : Google Scholar : PubMed/NCBI
27	Maeda K, Ding Q, Yoshimitsu M, Kuwahata T, Miyazaki Y, Tsukasa K, Hayashi T, Shinchi H, Natsugoe S and Takao S: CD133 modulate HIF-1α expression under hypoxia in EMT phenotype pancreatic cancer stem-like cells. Int J Mol Sci. 17:10252016. View Article : Google Scholar
28	Bar EE, Lin A, Mahairaki V, Matsui W and Eberhart CG: Hypoxia increases the expression of stem-cell markers and promotes clonogenicity in glioblastoma neurospheres. Am J Pathol. 177:1491–1502. 2010. View Article : Google Scholar : PubMed/NCBI
29	Conley SJ, Gheordunescu E, Kakarala P, Newman B, Korkaya H, Heath AN, Clouthier SG and Wicha MS: Antiangiogenic agents increase breast cancer stem cells via the generation of tumor hypoxia. Proc Natl Acad Sci U S A. 109:2784–2789. 2012. View Article : Google Scholar : PubMed/NCBI
30	Guo Q, Lan F, Yan X, Xiao Z, Wu Y and Zhang Q: Hypoxia exposure induced cisplatin resistance partially via activating p53 and hypoxia inducible factor-1α in non-small cell lung cancer A549 cells. Oncol Lett. 16:801–808. 2018.PubMed/NCBI
31	Yang ZL, Tian W, Wang Q, Zhao Y, Zhang YL, Tian Y, Tang YX, Wang SJ, Liu Y, Ni QQ, et al: Oxygen-evolving mesoporous organosilica coated prussian blue nanoplatform for highly efficient photodynamic therapy of tumors. Adv Sci (Weinh). 5:17008472018. View Article : Google Scholar : PubMed/NCBI
32	Zhao H, Wu Y, Chen Y and Liu H: Clinical significance of hypoxia-inducible factor 1 and VEGF-A in osteosarcoma. Int J Clin Oncol. 20:1233–1243. 2015. View Article : Google Scholar : PubMed/NCBI
33	Manresa MC and Taylor CT: Hypoxia inducible factor (HIF) hydroxylases as regulators of intestinal epithelial barrier function. Cell Mol Gastroenterol Hepatol. 3:303–315. 2017. View Article : Google Scholar : PubMed/NCBI
34	Gorga A, Rindone G, Regueira M, Riera MF, Pellizzari EH, Cigorraga SB, Meroni SB and Galardo MN: HIF involvement in the regulation of rat Sertoli cell proliferation by FSH. Biochem Biophys Res Commun. 502:508–514. 2018. View Article : Google Scholar : PubMed/NCBI
35	Bao L, Chen Y, Lai HT, Wu SY, Wang JE, Hatanpaa KJ, Raisanen JM, Fontenot M, Lega B, Chiang CM, et al: Methylation of hypoxia-inducible factor (HIF)-1α by G9a/GLP inhibits HIF-1 transcriptional activity and cell migration. Nucleic Acids Res. 46:6576–6591. 2018. View Article : Google Scholar : PubMed/NCBI
36	Guo M, Cai C, Zhao G, Qiu X, Zhao H, Ma Q, Tian L, Li X, Hu Y, Liao B, et al: Hypoxia promotes migration and induces CXCR4 expression via HIF-1α activation in human osteosarcoma. PLoS One. 9:e905182014. View Article : Google Scholar : PubMed/NCBI
37	Dodd KM, Yang J, Shen MH, Sampson JR, Tee AR and KM D: mTORC1 drives HIF-1α and VEGF-A signalling via multiple mechanisms involving 4E-BP1, S6K1 and STAT3. Oncogene. 34:2239–2250. 2015. View Article : Google Scholar : PubMed/NCBI
38	Cornu M, Albert V, Hall MN and M C: mTOR in aging, metabolism, and cancer. Curr Opin Genet Dev. 23:53–62. 2013. View Article : Google Scholar : PubMed/NCBI
39	Verheul HM, Salumbides B, Van Erp K, Hammers H, Qian DZ, Sanni T, Atadja P and Pili R: Combination strategy targeting the hypoxia inducible factor-1 alpha with mammalian target of rapamycin and histone deacetylase inhibitors. Clin Cancer Res. 14:3589–3597. 2008. View Article : Google Scholar : PubMed/NCBI
40	Hou H, Miao J, Cao R, Han M, Sun Y, Liu X and Guo L: Rapamycin ameliorates experimental autoimmune encephalomyelitis by suppressing the mTOR-STAT3 pathway. Neurochem Res. 42:2831–2840. 2017. View Article : Google Scholar : PubMed/NCBI