Hyperthermia reduces migration of osteosarcoma by suppression of autocrine motility factor

Nakajima,Kosei; Yanagawa,Takashi; Watanabe,Hideomi; Takagishi,Kenji

doi:10.3892/or.2012.2066

Hyperthermia reduces migration of osteosarcoma by suppression of autocrine motility factor

Authors:
- Kosei Nakajima
- Takashi Yanagawa
- Hideomi Watanabe
- Kenji Takagishi
View Affiliations

Affiliations: Department of Orthopedic Surgery, Graduate School of Medicine, Gunma University, Maebashi, Gunma, Japan, Department of Physical Therapy, Graduate School of Health Science, Gunma University, Maebashi, Gunma, Japan
Published online on: October 1, 2012 https://doi.org/10.3892/or.2012.2066
Pages: 1953-1958
Copyright: © Nakajima et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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

Autocrine motility factor (AMF) plays an important role in the development of metastasis by regulating tumor cell motility. The expression of AMF is associated with metastasis in malignant musculoskeletal tumors including osteosarcoma. Recent studies indicated that hyperthermia contributes to the improvement of the prognosis of patients with soft tissue sarcomas; however, few reports have evaluated the impact of hyperthermia on tumor cell motility, which is an important factor of metastasis. The purpose of this study was to evaluate the effect of hyperthermia with or without heat shock protein (HSP) inhibitors on the motility and AMF expression in an osteosarcoma cell line. Hyperthermia was carried out at 41˚C for 24 h. According to microarray results, HSP90, HSP70 and HSP27 expression was upregulated in osteosarcoma cells under hyperthermia. The intracellular, secreted AMF, mRNA of AMF and cell motility were evaluated by western blotting, ELISA, RT-PCR, wound healing and phagokinetic track assays, respectively. The protein secretion and mRNA levels of AMF and tumor cell motility were significantly decreased by hyperthermia. Of note, the downregulated AMF expression and motility were recovered by the addition of an HSP27 inhibitor. By contrast, the HSP90 and HSP70/72/105 inhibitors had no effect on AMF expression and motility downregulated by hyperthermia. In conclusion, hyperthermia reduced AMF expression and tumor cell motility via HSP27 and may therefore be applied as osteosarcoma treatment.

View Figures

View References

1	Bielack SS, Kempf-Bielack B, Delling G, 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
2	Whelan JS, Jinks RC, McTiernan A, et al: Survival from high-grade localised extremity osteosarcoma: combined results and prognostic factors from three European Osteosarcoma Intergroup randomised controlled trials. Ann Oncol. Oct 19–2011.(Epub ahead of print). View Article : Google Scholar
3	Bacci G, Longhi A, Versari M, Mercuri M, Briccoli A and Picci P: Prognostic factors for osteosarcoma of the extremity treated with neoadjuvant chemotherapy: 15-year experience in 789 patients treated at a single institution. Cancer. 106:1154–1161. 2006.PubMed/NCBI
4	Ferrari S, Smeland S, Mercuri M, et al: Neoadjuvant chemotherapy with high-dose Ifosfamide, high-dose methotrexate, cisplatin, and doxorubicin for patients with localized osteosarcoma of the extremity: a joint study by the Italian and Scandinavian Sarcoma Groups. J Clin Oncol. 23:8845–8852. 2005. View Article : Google Scholar
5	Mirabello L, Troisi RJ and Savage SA: Osteosarcoma incidence and survival rates from 1973 to 2004: data from the Surveillance, Epidemiology, and End Results Program. Cancer. 115:1531–1543. 2009. View Article : Google Scholar : PubMed/NCBI
6	Tsutsumi S, Yanagawa T, Shimura T, et al: Regulation of cell proliferation by autocrine motility factor/phosphoglucose isomerase signaling. J Biol Chem. 278:32165–32172. 2003. View Article : Google Scholar : PubMed/NCBI
7	Silletti S, Watanabe H, Hogan V, Nabi IR and Raz A: Purification of B16-F1 melanoma autocrine motility factor and its receptor. Cancer Res. 51:3507–3511. 1991.PubMed/NCBI
8	Watanabe H, Kanbe K and Chigira M: Differential purification of autocrine motility factor derived from a murine protein-free fibrosarcoma. Clin Exp Metastasis. 12:155–163. 1994. View Article : Google Scholar : PubMed/NCBI
9	Funasaka T, Haga A, Raz A and Nagase H: Tumor autocrine motility factor is an angiogenic factor that stimulates endothelial cell motility. Biochem Biophys Res Commun. 284:1116–1125. 2001. View Article : Google Scholar : PubMed/NCBI
10	Funasaka T, Haga A, Raz A and Nagase H: Autocrine motility factor secreted by tumor cells upregulates vascular endothelial growth factor receptor (Flt-1) expression in endothelial cells. Int J Cancer. 101:217–223. 2002. View Article : Google Scholar
11	Tsutsumi S, Hogan V, Nabi IR and Raz A: Overexpression of the autocrine motility factor/phosphoglucose isomerase induces transformation and survival of NIH-3T3 fibroblasts. Cancer Res. 63:242–249. 2003.PubMed/NCBI
12	Haga A, Funasaka T, Niinaka Y, Raz A and Nagase H: Autocrine motility factor signaling induces tumor apoptotic resistance by regulations Apaf-1 and Caspase-9 apoptosome expression. Int J Cancer. 107:707–714. 2003. View Article : Google Scholar : PubMed/NCBI
13	Watanabe H, Takehana K, Date M, Shinozaki T and Raz A: Tumor cell autocrine motility factor is the neuroleukin/phosphohexose isomerase polypeptide. Cancer Res. 56:2960–2963. 1996.PubMed/NCBI
14	Faik P, Walker JI, Redmill AA and Morgan MJ: Mouse glucose-6-phosphate isomerase and neuroleukin have identical 3′ sequences. Nature. 332:455–457. 1988.
15	Niinaka Y, Paku S, Haga A, Watanabe H and Raz A: Expression and secretion of neuroleukin/phosphohexose isomerase/maturation factor as autocrine motility factor by tumor cells. Cancer Res. 58:2667–2674. 1998.PubMed/NCBI
16	Yanagawa T, Watanabe H, Takeuchi T, Fujimoto S, Kurihara H and Takagishi K: Overexpression of autocrine motility factor in metastatic tumor cells: possible association with augmented expression of KIF3A and GDI-beta. Lab Invest. 84:513–522. 2004. View Article : Google Scholar : PubMed/NCBI
17	Yanagawa T, Funasaka T, Tsutsumi S, Raz T, Tanaka N and Raz A: Differential regulation of phosphoglucose isomerase/autocrine motility factor activities by protein kinase CK2 phosphorylation. J Biol Chem. 280:10419–10426. 2005. View Article : Google Scholar
18	Baumann M, Kappl A, Lang T, Brand K, Siegfried W and Paterok E: The diagnostic validity of the serum tumor marker phosphohexose isomerase (PHI) in patients with gastrointestinal, kidney, and breast cancer. Cancer Invest. 8:351–356. 1990. View Article : Google Scholar : PubMed/NCBI
19	Filella X, Molina R, Jo J, Mas E and Ballesta AM: Serum phosphohexose isomerase activities in patients with colorectal cancer. Tumour Biol. 12:360–367. 1991. View Article : Google Scholar : PubMed/NCBI
20	Bodansky O: Serum phosphohexose isomerase in cancer. II. As an index of tumor growth in metastatic carcinoma of the breast. Cancer. 7:1200–1226. 1954. View Article : Google Scholar : PubMed/NCBI
21	Patel PS, Raval GN, Rawal RM, et al: Comparison between serum levels of carcinoembryonic antigen, sialic acid and phosphohexose isomerase in lung cancer. Neoplasma. 42:271–274. 1995.PubMed/NCBI
22	Dobashi Y, Watanabe H, Matsubara M, et al: Autocrine motility factor/glucose-6-phosphate isomerase is a possible predictor of metastasis in bone and soft tissue tumours. J Pathol. 208:44–53. 2006. View Article : Google Scholar : PubMed/NCBI
23	Niinaka Y, Harada K, Fujimuro M, et al: Silencing of autocrine motility factor induces mesenchymal-to-epithelial transition and suppression of osteosarcoma pulmonary metastasis. Cancer Res. 70:9483–9493. 2010. View Article : Google Scholar : PubMed/NCBI
24	Issels RD, Lindner LH, Verweij J, et al: Neo-adjuvant chemotherapy alone or with regional hyperthermia for localised high-risk soft-tissue sarcoma: a randomised phase 3 multicentre study. Lancet Oncol. 11:561–570. 2010. View Article : Google Scholar
25	Alcaide M, Ramírez-Santillán C, Feito MJ, et al: In vitro evaluation of glass-glass ceramic thermoseed-induced hyperthermia on human osteosarcoma cell line. J Biomed Mater Res A. 100:64–71. 2012. View Article : Google Scholar : PubMed/NCBI
26	Trieb K, Blahovec H and Kubista B: Effects of hyperthermia on heat shock protein expression, alkaline phosphatase activity and proliferation in human osteosarcoma cells. Cell Biochem Funct. 25:669–672. 2007. View Article : Google Scholar : PubMed/NCBI
27	Shui C and Scutt A: Mild heat shock induces proliferation, alkaline phosphatase activity, and mineralization in human bone marrow stromal cells and Mg-63 cells in vitro. J Bone Miner Res. 16:731–741. 2001. View Article : Google Scholar
28	Shido Y, Nishida Y, Suzuki Y, Kobayashi T and Ishiguro N: Targeted hyperthermia using magnetite cationic liposomes and an alternating magnetic field in a mouse osteosarcoma model. J Bone Joint Surg Br. 92:580–585. 2010. View Article : Google Scholar
29	Bagatell R, Beliakoff J, David CL, Marron MT and Whitesell L: Hsp90 inhibitors deplete key anti-apoptotic proteins in pediatric solid tumor cells and demonstrate synergistic anticancer activity with cisplatin. Int J Cancer. 113:179–188. 2005. View Article : Google Scholar : PubMed/NCBI
30	Shin KD, Lee MY, Shin DS, et al: Blocking tumor cell migration and invasion with biphenyl isoxazole derivative KRIBB3, a synthetic molecule that inhibits Hsp27 phosphorylation. J Biol Chem. 280:41439–41448. 2005. View Article : Google Scholar : PubMed/NCBI
31	Yokota S, Kitahara M and Nagata K: Benzylidene lactam compound, KNK437, a novel inhibitor of acquisition of thermotolerance and heat shock protein induction in human colon carcinoma cells. Cancer Res. 60:2942–2948. 2000.PubMed/NCBI
32	Bagatell R, Gore L, Egorin MJ, et al: Phase I pharmacokinetic and pharmacodynamic study of 17-N-allylamino-17-demethoxygeldanamycin in pediatric patients with recurrent or refractory solid tumors: a pediatric oncology experimental therapeutics investigators consortium study. Clin Cancer Res. 13:1783–1788. 2007. View Article : Google Scholar
33	Koga F, Kihara K and Neckers L: Inhibition of cancer invasion and metastasis by targeting the molecular chaperone heat-shock protein 90. Anticancer Res. 29:797–807. 2009.PubMed/NCBI
34	Sato T, Sawaji Y, Matsui N, et al: Heat shock suppresses membrane type 1-matrix metalloproteinase production and progelatinase A activation in human fibrosarcoma HT-1080 cells and thereby inhibits cellular invasion. Biochem Biophys Res Commun. 265:189–193. 1999. View Article : Google Scholar
35	Funasaka T, Hu H, Yanagawa T, Hogan V and Raz A: Down-regulation of phosphoglucose isomerase/autocrine motility factor results in mesenchymal-to-epithelial transition of human lung fibrosarcoma cells. Cancer Res. 67:4236–4243. 2007. View Article : Google Scholar : PubMed/NCBI
36	Funasaka T, Hogan V and Raz A: Phosphoglucose isomerase/autocrine motility factor mediates epithelial and mesenchymal phenotype conversions in breast cancer. Cancer Res. 69:5349–5356. 2009. View Article : Google Scholar
37	Kugler W, Breme K, Laspe P, et al: Molecular basis of neurological dysfunction coupled with haemolytic anaemia in human glucose-6-phosphate isomerase (GPI) deficiency. Hum Genet. 103:450–454. 1998. View Article : Google Scholar : PubMed/NCBI
38	Yanagawa T, Funasaka T, Tsutsumi S, Hu H, Watanabe H and Raz A: Regulation of phosphoglucose isomerase/autocrine motility factor activities by the poly(ADP-ribose) polymerase family-14. Cancer Res. 67:8682–8689. 2007. View Article : Google Scholar : PubMed/NCBI
39	Nakano H, Tateishi A, Miki H, et al: Hyperthermic isolated regional perfusion for the treatment of osteosarcoma in the lower extremity. Am J Surg. 178:27–32. 1999. View Article : Google Scholar : PubMed/NCBI
40	Williams RR, Hassan-Walker AF, Lavender FL, Morgan M, Faik P and Ragoussis J: The minisatellite of the GPI/AMF/NLK/MF gene: interspecies conservation and transcriptional activity. Gene. 269:81–92. 2001. View Article : Google Scholar : PubMed/NCBI
41	Funasaka T, Yanagawa T, Hogan V and Raz A: Regulation of phosphoglucose isomerase/autocrine motility factor expression by hypoxia. FASEB J. 19:1422–1430. 2005. View Article : Google Scholar : PubMed/NCBI
42	Khalil AA, Kabapy NF, Deraz SF and Smith C: Heat shock proteins in oncology: diagnostic biomarkers or therapeutic targets? Biochim Biophys Acta. 1816:89–104. 2011.PubMed/NCBI
43	Jego G, Hazoumé A, Seigneuric R and Garrido C: Targeting heat shock proteins in cancer. Cancer Lett. Nov 13–2010.(Epub ahead of print).
44	Uozaki H, Horiuchi H, Ishida T, Iijima T, Imamura T and Machinami R: Overexpression of resistance-related proteins (metallothioneins, glutathione-S-transferase pi, heat shock protein 27, and lung resistance-related protein) in osteosarcoma. Relationship with poor prognosis. Cancer. 79:2336–2344. 1997. View Article : Google Scholar
45	Moon A, Bacchini P, Bertoni F, et al: Expression of heat shock proteins in osteosarcomas. Pathology. 42:421–425. 2010. View Article : Google Scholar : PubMed/NCBI
46	Têtu B, Lacasse B, Bouchard HL, Lagacé R, Huot J and Landry J: Prognostic influence of HSP-27 expression in malignant fibrous histiocytoma: a clinicopathological and immunohistochemical study. Cancer Res. 52:2325–2328. 1992.PubMed/NCBI