Human MG-63 osteosarcoma cells responses to long and short term hyper- and hypothermia stress
DOI:
https://doi.org/10.11113/mjfas.v16n5.1344Keywords:
hyperthermia, hypothermia, osteosarcoma, heat shock proteins, cold shock proteinsAbstract
Hyper- and hypothermia are utilized as treatment modalities in cancer treatment or as a protection against ischemia-reperfusion induced cell damage. The under-lying mechanism of hyper- and hypothermia, on cell death in osteosarcoma cells are not well understood. The aim of this study is to investigate the short- and long-term effects of various severities of hyper- and hypothermia on osteoblast-like osteosarcoma cells (MG-63). MG-63 cells were treated with mild and severe hyper- and hypothermia for short, medium and long-term periods. Severe hypothermia and hyperthermia showed a time-dependent toxicity; hence viability was reduced in a significant manner at all time points and the cells were undergoing apoptosis. Mild hypothermia, on the other hand, showed a protective effect and long term exposure increased the cell viability. Severe hyperthermia induced significant DNA damage at all time points. Caspase 3/7 activity showed a significant increase at 1 h of severe hyperthermia and 72 h of severe hypothermia (p<0.05). Hsp90 expression was significantly increased at 72 h of mild hyperthermia (p<0.01), whereas Hsp70 showed a significant increase after 24 and 72 h (p<0.01 and p<0.001). Hsp27 mRNA was increased significantly at 24 h only under mild hyperthermia (p<0.01). In conclusion, hyperthermia especially severe hyperthermia induced cellular stress in MG-63 cells leading to apoptosis. Hypothermia, on the other hand caused severe cell stress only when the cells were challenged for a prolonged period with severe low temperatures.
References
Matsuzaki Y, Edagawa M, Shimizu T, Hara M, Tomita M, Ayabe T, Onitsuka T (2004) Intrapleural hyperthermic perfusion with chemotherapy increases apoptosis in malignant pleuritis. Ann Thorac Surg. 78:1769-1772. http://dx.doi.org/10.1016/j.athoracsur.2004.05.025.
Hurwitz MD (2010) Today's thermal therapy: Not your father's hyperthermia: Challenges and opportunities in application of hyperthermia for the 21st century cancer patient. Am. J. Clin. Oncol. 33:96-100. doi: 10.1097/COC.0b013e3181817a75.
Asfour HZ, AlQahtani MH, Balto KA, ElAssouli SM (2011) Heat shock proteins, estrogen and progesterone receptors expression in breast carcinomas. J Med Med Sci. 2: 977-984
Zeng Y, Kulkarni P, Inoue T, Getzenberg RH (2009) Down‐regulating cold shock protein genes impairs cancer cell survival and enhances chemosensitivity. J Cell Biochem. 107: 179-188. doi: 10.1002/jcb.22114.
Kalamida D, Karagounis IV, Mitraka SA, Kalamida S, Giatromanolaki A, Koukourakis MI (2015) Fever-Range Hyperthermia vs. Hypothermia Effect on Cancer Cell Viability, Proliferation and HSP90 Expression. PLoSONE 10(1):e0116021. doi:10.1371/journal.pone.0116021
Hogendoorn PCW, Athanasou N, Bielack S, De Alava E, Dei Tos AP, Ferrari S, Gelderblom H, Grimer R, Hall KS, Hassan B, Jurgens H (2010) Bone sarcomas: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol. 21:204-213. doi: 10.1093/annonc/mdq223.
López-Martínez JJ, García-Sandoval PP, Fernández-Hernández JA, Valcárcel-Díaz, A (2012) Functionality and osteointegration of bone allografts in long bone osteosarcomas. Acta Ortop Mex. 26: 30-34.
Strauss SJ, Ng T, Mendoza-Naranjo A, Whelan J, Sorensen PH (2010) Understanding micrometastatic disease and Anoikis resistance in ewing family of tumors and osteosarcoma. Oncologist.15:627-635. doi: 10.1634/theoncologist.2010-0093
Trieb K, Blahovec H, Kubista B (2007) Effects of hyperthermia on heat shock protein expression, alkaline phosphatase activity and proliferation in human osteosarcoma cells. Cell Biochem Funct. 25:669-672. doi: 10.1002/cbf.1371
Rong Y, Mack P (2000) Apoptosis induced by hyperthermia in Dunn osteosarcoma cell line in vitro. Int J Hyperthermia. 16: 19-27
Kanamori M, Ohmori K, Yasuda T, Yudoh K (2003) Effects of hyperthermia and differentiation on cultured Dunn osteosarcoma cells. Cancer Detect. Prev. 27:76-81
Andresen M, Gazmuri JT, Marín A, Regueira T, Rovegno M (2015) Therapeutic hypothermia for acute brain injuries. SJTREM. 23:42. doi: 10.1186/s13049-015-0121-3
Fujita J (1999) Cold shock response in mammalian cells. J Mol Microbiol Biote chnol. 1:243-255
Sonna LA, Fujita J, Gaffin SL, Lilly CM, (2002) Invited review: effects of heat and cold stress on mammalian gene expression. J Appl Physiol. 92:1725-1742. doi: 10.1152/japplphysiol.01143.2001
Sonna LA, Kuhlmeier MM, Carter HC, Hasday JD, Lilly CM, Fairchild KD (2006) Effect of moderate hypothermia on gene expression by THP-1 cells: a DNA microarray study. Physiol Genomics. 26: 91-98. doi: 10.1152/physiolgenomics.00296.2005
Nishiyama H, Danno S, Kaneko Y, Itoh K, Yokoi H, Fukumoto M, Okuno H, Millan JL, Matsuda T, Yoshida O, Fujita J (1998) Decreased expression of cold-inducible RNA-binding protein (CIRP) in male germ cells at elevated temperature. Am J Pathol. 289-296
Aisha MD, Nor-Ashikin MNK, Sharaniza AB, Nawawi HM, Kapitonova MY, Froemming GRA (2014) Short-term moderate hypothermia stimulates alkaline phosphatase activity and osteocalcin expression in osteoblasts by upregulating Runx2 and Osterix in vitro. Exp Cell Res. 326:46-56.doi: 10.1016/j.yexcr.2014.06.003. Epub 2014 Jun 11.
Aisha, M. D., Kapitonova, M., Nor-Ashikin, M. N. K., Sharaniza, A. R., Nawawi, H., & Froemming, G. R. A (2012). Acute moderate and severe hypo-and hyperthermia down-regulate gene expression of molecular chaperones and inflammatory cytokines in osteoblast cells. Bone, 50, S81.
Aisha, M. D., Ab Rahim, S., Nawawi, H., Nor-Ashikin, M. N., & Froemming, G. R. A (2012) Short Term Effects Of Moderate And Severe Hypo-And Hyperthermia On Normal Human Osteoblast Cellular Gene Function. In Osteoporosis International (Vol. 23, pp. S760-S760). 236 GRAYS INN RD, 6TH Floor, London WC1X 8HL, England: Springer London LTD.
Mosmann, T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 65:55-63.
Singh, Narendra P, Michael T McCoy, Raymond R Tice, and Edward L Schneider. 1988. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res. 175:184-191.
Al-Salahi OS, Ji D, Majid AM, Kit-Lam C, Abdullah WZ, Zaki A, Jamal Din SK, Yusoff NM, Majid AS. (2014) Anti-tumor activity of Eurycoma longifolia root extracts against K-562 cell line: in vitro and in vivo study. PloS one. 9:e83818. doi: 10.1371/journal.pone.0083818. eCollection 2014.
Nassar ZD, Aisha AF, Al Suede FS, Abdul Majid AS, Abdul Majid AM (2012) In vitro antimetastatic activity of koetjapic acid against breast cancer cells. Biol Pharm Bull. 35:503-508.
Desjardins P, Hansen JB, Allen M (2009 Microvolume protein concentration determination using the NanoDrop 2000c spectrophotometer. J Vis Exp.4:e1610. doi: 10.3791/1610.
Garcia MP, Cavalheiro JRT, Fernandes MH (2012) Acute and long-term effects of hyperthermia in B16-F10 melanoma cells. PloS one. 7. doi.org/10.1371/journal.pone.0035489
Luchetti F, Mannello F, Canonico B, Battistelli M, Burattini S, Falcieri E, Papa S (2004) Integrin and cytoskeleton behaviour in human neuroblastoma cells during hyperthermia-related apoptosis. Apoptosis. 9:635-648. doi:10.1023/B:APPT.0000038043.03799.6f.
O'neill KL, Fairbairn DW, Smith MJ, Poe BS (1998) Critical parameters influencing hyperthermia-induced apoptosis in human lymphoid cell lines. Apoptosis. 3:369-375.
Elengoe A, Hamdan S (2014) Evaluation of hyperthermia effect on cell viability using Crystal violet staining, LDH and Trypan blue assays. Adv. Environ. Biol.744-748
Pawlik A, Nowak JM, Grzanka D, Gackowska L, Michalkiewicz J, Grzanka A (2013) Hyperthermia induces cytoskeletal alterations and mitotic catastrophe in p53-deficient H1299 lung cancer cells. Acta histochem. 115:8-15. doi: 10.1016/j.acthis.2012.02.006
Fukao H, Ikeda M, Ichikawa T, Inufusa H, Okada K, Ueshima S, Matsuo O (2000) Effect of hyperthermia on the viability and the fibrinolytic potential of human cancer cell lines. Clin Chim Acta, 296:17-33
Matijasevic, Z (2002) Selective protection of non-cancer cells by hypothermia. Anticancer research. 22: 3267-3272.
Rainey JB, Davies PW, Williamson RC (1987) Effect of hypothermia on intestinal adaptation and carcinogenesis in the rat. Br J Cancer. 55: 265-268
Sunley K, Tharmalingam T, Butler M (2008) CHO cells adapted to hypothermic growth produce high yields of recombinant β‐interferon. Biotechnol Prog. 24:898-906. doi: 10.1002/btpr.9.
Shi M, Xie Z, Yu M, Shen B, Guo N, (2005) Controlled growth of Chinese hamster ovary cells and high expression of antibody-IL-2 fusion proteins by temperature manipulation. Biotechnol Lett. 27:1879-1884. doi:10.1007/s10529-005-3897-9
Van Rijn J, Van den Berg J, Kipp JB, Schamhart DH, Van Wijk R (1985) Effect of hypothermia on cell kinetics and response to hyperthermia and X rays. Radiat Res. 101: 292-305. doi: 10.2307/3576395
Takahashi A, Ohnishi T (2005) Does γH2AX foci formation depend on the presence of DNA double strand breaks?. Cancer Lett. 229:171-179. http://dx.doi.org/10.1016/j.canlet.2005.07.016
Roti Roti JL, Pandita RK, Mueller JD, Novak P, Moros EG, Laszlo A (2010) Severe, short-duration (0–3 min) heat shocks (50–52 C) inhibit the repair of DNA damage Int J Hyperthermia. 26: 67-78. doi: 10.3109/02656730903417947
Milhas D, Cuvillier O, Therville N, Clavé P, Thomsen M, Levade T, Benoist H, Ségui B (2005) Caspase-10 triggers Bid cleavage and caspase cascade activation in FasL-induced apoptosis. J. Biol. Chem. 280:19836-19842. doi: 10.1074/jbc.M414358200
Schmitt E, Gehrmann M, Brunet M, Multhoff G, Garrido C (2007) Intracellular and extracellular functions of heat shock proteins: repercussions in cancer therapy. J Leukoc Biol. 81:15-27. doi:10.1189/jlb.0306167
Samali A, Robertson JD, Peterson E, Manero F, van Zeijl L, Paul C, Cotgreave IA, Arrigo AP, Orrenius S (2001) Hsp27 protects mitochondria of thermotolerant cells against apoptotic stimuli. Cell Stress & Chaperones. 6: 49-58
Concannon CG, Orrenius STEN, Samali AFSHIN (2001) Hsp27 inhibits cytochrome c-mediated caspase activation by sequestering both pro-caspase-3 and cytochrome c. Gene Expr. 9:195-201. DOI: https://doi.org/10.3727/000000001783992605
Nakajima K, Yanagawa T, Watanabe H, Takagishi K (2012) Hyperthermia reduces migration of osteosarcoma by suppression of autocrine motility factor. Oncol Rep. 28:1953-1958. doi: 10.3892/or.2012.2066
Trieb K, Lechleitner T, Lang S, Windhager R, Kotz R, Dirnhofer S. 1998. Heat shock protein 72 expression in osteosarcomas correlates with good response to neoadjuvant chemotherapy. Hum Pathol. 29: 1050-1055
Uozaki H, Ishida T, Kakiuchi C, Horiuchi H, Gotoh T, Iijima T, Imamura T, Machinami R (2000) Expression of heat shock proteins in osteosarcoma and its relationship to prognosis. Pathol Res Pract. 196:665-673. doi:10.1016/S0344-0338(00)80118-1
Morii T, Ohtsuka K, Ohnishi H, Mochizuki K, Satomi K (2010) Inhibition of heat-shock protein 27 expression eliminates drug resistance of osteosarcoma to zoledronic acid. Anticancer Res, 30: 3565-3571.
Lieonart ME (2010) A new generation of proto-oncogenes: cold-inducible RNA binding proteins. Biochim. Biophys. Acta (BBA)-Reviews on Cancer. 1805:43-52. http://dx.doi.org/10.1016/j.bbcan.2009.11.001
Dresios J, Aschrafi A, Owens GC, Vanderklish PW, Edelman GM, Mauro VP (2005) Cold stress-induced protein Rbm3 binds 60S ribosomal subunits, alters microRNA levels, and enhances global protein synthesis. Proc. Natl. Acad. Sci. U.S.A.102:1865-1870. doi: 10.1073/pnas.0409764102
Jögi A, Brennan DJ, Rydén L, Magnusson K, Fernö M, Stål O, Borgquist S, Uhlen M, Landberg G, Påhlman S, Pontén F (2009) Nuclear expression of the RNA-binding protein RBM3 is associated with an improved clinical outcome in breast cancer.Modern Patho. 22:1564-1574. doi: 10.1038/modpathol.2009.124.
Ehlen A, Brennan DJ, Nodin B, O'Connor DP, Eberhard J, Alvarado-Kristensson M, Jeffrey IB, Manjer J, Brandstedt J, Uhlén M, Pontén F (2010) Expression of the RNA-binding protein RBM3 is associated with a favourable prognosis and cisplatin sensitivity in epithelial ovarian cancer. J Transl Med. 8:10-1186. doi: 10.1186/1479-5876-8-78
Jonsson L, Gaber A, Ulmert D, Uhlén M, Bjartell A, Jirström K (2011) High RBM3 expression in prostate cancer independently predicts a reduced risk of biochemical recurrence and disease progression. Diagn Pathol 6:91. doi: 10.1186/1746-1596-6-91
Zeng Y, Wodzenski D, Gao D, Shiraishi T, Terada N, Li Y, Vander Griend DJ, Luo J, Kong C, Getzenberg RH, Kulkarni P (2013) Stress-response protein RBM3 attenuates the stem-like properties of prostate cancer cells by interfering with CD44 variant splicing. Cancer Res. 73:4123-4133. doi: 10.1158/0008-5472.CAN-12-1343.
