Cisplatin‑resistant osteosarcoma cells possess cancer stem cell properties in a mouse model

Yang,Jian; Guo,Weichun; Wang,Lu; Yu,Ling; Mei,Hongjun; Fang,Shuo; Ji,Peng; Liu,Yang; Liu,Gaiwei; Song,Qi

doi:10.3892/ol.2016.4956

Cisplatin‑resistant osteosarcoma cells possess cancer stem cell properties in a mouse model

Authors:
- Jian Yang
- Weichun Guo
- Lu Wang
- Ling Yu
- Hongjun Mei
- Shuo Fang
- Peng Ji
- Yang Liu
- Gaiwei Liu
- Qi Song
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Affiliations: Department of Orthopedics, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China, Department of Gynecology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China, Department of Orthopedics, Wuhan No. 5 Hospital, Wuhan, Hubei 430050, P.R. China
Published online on: August 5, 2016 https://doi.org/10.3892/ol.2016.4956
Pages: 2599-2605
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Osteosarcoma is the most common malignancy of the bones, and although advances in chemotherapy and surgery had been achieved in recent years, the long‑term survival rate has reached a plateau. The main reason for this is the aggressive malignant potential and poor response of the disease to chemotherapy. However, several studies have found that tumor resistance is associated with cancer stem cells (CSCs). To address this issue, in the present study, osteosarcoma cells were treated with specially designated concentrations of cisplatin (CDDP) in a mouse model. Hematoxylin and eosin staining analyses were performed to assess tissue structure, in vivo passaging and CDDP treatment. Drug resistance genes and well‑established stemness genes were detected by quantitative polymerase chain reaction. A serum‑starved sphere formation assay was adopted to evaluate the ability to generate spherical clones and flow cytometry as used to test the expression of the cluster of differentiation 117 and Stro‑1 surface markers, known as markers of CSCs. It was found that CDDP could induce an effect of resistance in the osteosarcoma cells, which possessed cancer stem CSC properties, as shown by the elevated expression of CSC marker genes and the higher expression of the cluster of differentiation 117 and Stro‑1 surface markers. Moreover, the cells that dissociated from the tumor tissues exhibited an increased ability to form sarcospheres. The results of this study provided a significant correlation between resistance and CSCs, and revealed a clue indicating that osteosarcoma recurrence is likely to be associated with CSCs.

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1	Bielack SS, Kempf-Bielack B, Delling G, Exner GU, Flege S, Helmke K, Kotz R, Salzer-Kuntschik M, Werner M, Winkelmann W, et al: Prognostic factors in high-grade osteosarcoma of the extremities or trunk: An analysis of 1,702 patients treated on neoadjuvant cooperative osteosarcoma study group protocols. J Clin Oncol. 20:776–790. 2002. View Article : Google Scholar : PubMed/NCBI
2	Cormier JN and Pollock RE: Soft tissue sarcomas. CA Cancer J Clin. 54:94–109. 2004. View Article : Google Scholar : PubMed/NCBI
3	Luetke A, Meyers PA, Lewis I and Juergens H: Osteosarcoma treatment-where do we stand? A state of the art review. Cancer Treat Rev. 40:523–532. 2014. View Article : Google Scholar : PubMed/NCBI
4	Janeway KA and Grier HE: Sequelae of osteosarcoma medical therapy: A review of rare acute toxicities and late effects. Lancet Oncol. 11:670–678. 2010. View Article : Google Scholar : PubMed/NCBI
5	Picci P, Ferrari S, Bacci G and Gherlinzoni F: Treatment recommendations for osteosarcoma and adult soft tissue sarcomas. Drugs. 47:82–92. 1994. View Article : Google Scholar : PubMed/NCBI
6	Meyers PA: Muramyl tripeptide (mifamurtide) for the treatment of osteosarcoma. Expert Rev Anticancer Ther. 9:1035–1049. 2009. View Article : Google Scholar : PubMed/NCBI
7	Gorlick R, Anderson P, Andrulis I, Arndt C, Beardsley GP, Bernstein M, Bridge J, Cheung NK, Dome JS, Ebb D, et al: Biology of childhood osteogenic sarcoma and potential targets for therapeutic development: Meeting summary. Clin Cancer Res. 9:5442–5453. 2003.PubMed/NCBI
8	Bonnet D and Dick JE: Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med. 3:730–737. 1997. View Article : Google Scholar : PubMed/NCBI
9	Park CY, Tseng D and Weissman IL: Cancer stem cell-directed therapies: Recent data from the laboratory and clinic. Mol Ther. 17:219–230. 2009. View Article : Google Scholar : PubMed/NCBI
10	Dean M, Fojo T and Bates S: Tumour stem cells and drug resistance. Nat Rev Cancer. 5:275–284. 2005. View Article : Google Scholar : PubMed/NCBI
11	Korkaya H and Wicha MS: Selective targeting of cancer stem cells: A new concept in cancer therapeutics. BioDrugs. 21:299–310. 2007. View Article : Google Scholar : PubMed/NCBI
12	Ma S, Lee TK, Zheng BJ, Chan KW and Guan XY: CD133⁺ HCC cancer stem cells confer chemoresistance by preferential expression of the Akt/PKB survival pathway. Oncogene. 27:1749–1758. 2008. View Article : Google Scholar : PubMed/NCBI
13	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
14	Luu HH, Kang Q, Park JK, Si W, Luo Q, Jiang W, Yin H, Montag AG, Simon MA, Peabody TD, et al: An orthotopic model of human osteosarcoma growth and spontaneous pulmonary metastasis. Clin Exp Metastasis. 22:319–329. 2005. 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	Savage S.A..Mirabello L: Using epidemiology and genomics to understand osteosarcoma etiology. Sarcoma, 2011. 2011.548151
17	Ek ET, Dass CR and Choong PF: Commonly used mouse models of osteosarcoma. Crit Rev Oncol Hematol. 60:1–8. 2006. View Article : Google Scholar : PubMed/NCBI
18	Chen X, Li Y, Xiong K, Aizicovici S, Xie Y, Zhu Q, et al: Cancer gene therapy by direct tumor injections of a nonviral T7 vector encoding a thymidine kinase gene. Human Gene Ther. 1998.9(5): 729–36. View Article : Google Scholar
19	Crnalic S, Hakansson I, Boquist L, Lofvenberg R and Brostrom LA: A novel spontaneous metastasis model of human osteosarcoma developed using orthotopic transplantation of intact tumor tissue into tibia of nude mice. Clin Exp Metastasis. 1997.15(2): 164–72. View Article : Google Scholar : PubMed/NCBI
20	Vergote J, Moretti JL, de Vries EG and Garnier-Suillerot A: Comparison of the kinetics of active efflux of 99mTc-MIBI in cells with P-glycoprotein-mediated and multidrug-resistance protein-associated multidrug-resistance phenotypes. Eur J Biochem. 252:140–146. 1998. View Article : Google Scholar : PubMed/NCBI
21	Suto R, Abe Y, Nakamura M, Ohnishi Y, Yoshimura M, Lee YH, Imanishi T, Yamazaki H, Kijima H, Tokunaga T, et al: Multidrug resistance mediated by overexpression of P-glycoprotein in human osteosarcoma in vivo. Int J Oncol. 12:287–291. 1998.PubMed/NCBI
22	Chan HS, Grogan TM, Haddad G, DeBoer G and Ling V: P-glycoprotein expression: Critical determinant in the response to osteosarcoma chemotherapy. J Natl Cancer Inst. 89:1706–1715. 1997. View Article : Google Scholar : PubMed/NCBI
23	Burak Z, Moretti JL, Ersoy O, Sanli U, Kantar M, Tamgac F and Basdemir G: 99mTc-MIBI imaging as a predictor of therapy response in osteosarcoma compared with multidrug resistance-associated protein and P-glycoprotein expression. J Nucl Med. 44:1394–1401. 2003.PubMed/NCBI
24	Dutour A, Leclers D, Monteil J, Paraf F, Charissoux JL, Rousseau R and Rigaud M: Non-invasive imaging correlates with histological and molecular characteristics of an osteosarcoma model: Application for early detection and follow-up of MDR phenotype. Anticancer Res. 27:4171–4178. 2007.PubMed/NCBI
25	Beachy PA, Karhadkar SS and Berman DM: Tissue repair and stem cell renewal in carcinogenesis. Nature. 432:324–331. 2004. View Article : Google Scholar : PubMed/NCBI
26	Ravandi F, Burnett AK, Agura ED and Kantarjian HM: Progress in the treatment of acute myeloid leukemia. Cancer. 110:1900–1910. 2007. View Article : Google Scholar : PubMed/NCBI
27	Woodward WA and Sulman EP: Cancer stem cells: Markers or biomarkers? Cancer Metastasis Rev. 27:459–470. 2008. View Article : Google Scholar : PubMed/NCBI
28	Hay DC, Sutherland L, Clark J and Burdon T: Oct-4 knockdown induces similar patterns of endoderm and trophoblast differentiation markers in human and mouse embryonic stem cells. Stem Cells. 22:225–235. 2004. View Article : Google Scholar : PubMed/NCBI
29	Boyer LA, Lee TI, Cole MF, Johnstone SE, Levine SS, Zucker JP, Guenther MG, Kumar RM, Murray HL, Jenner RG, et al: Core transcriptional regulatory circuitry in human embryonic stem cells. Cell. 122:947–956. 2005. View Article : Google Scholar : PubMed/NCBI
30	Wei H, Zhao MQ, Dong W, Yang Y and Li JS: Expression of c-kit protein and mutational status of the c-kit gene in osteosarcoma and their clinicopathological significance. J Int Med Res. 36:1008–1014. 2008. View Article : Google Scholar : PubMed/NCBI