Critical Role for the Protons in FRTL-5 Thyroid Cells: Nuclear Sphingomyelinase Induced-Damage
Abstract
:1. Introduction
2. Results and Discussion
2.1. Proton Beams Promote Bax Expression in FRTL-5 Cells in Relation to Their Physiological State
2.2. Proton Beams Act on Nuclear Sphingomyelin Metabolism
SM Species | TSH−C | TSH−P | TSH+C | TSH+P |
---|---|---|---|---|
SM 18:1 12:0 | 0.7 (0.5–0.8) | 0,9 | 0.3 (0.2–0.6) | 0.15 |
SM 18:1 16:0 | 46 (41–58) | 50 | 64 (55–71) | 20 |
SM 18:1 18:1 | 5 (3.3–5.6) | 2,2 | 1.4 (1.1–1.5) | 1.4 |
SM 24:0 | 172 (160–203) | 184 | 188 (168–202) | 74 |
ceramide 18:1 16:0 | 117 (102–139) | 147 | 72 (62–68) | 25 |
ceramide 18:1 18:0 | 31 (23–36) | 48 | 2 (1.6–2.3) | 8 |
ceramide 18:1 20:0 | 45 (18–54) | 118 | 62 (40–71) | 27 |
3. Experimental Section
3.1. Reagents and Standards
3.2. Cell Cultures and Treatments
3.3. Lipid Extraction
3.4. Ultra Fast Liquid Chromatography Tandem Mass Spectrometry (UFLC-MS/MS)
3.5. Neutral-Sphingomyelinase Assay
3.6. Sphingomyelin-Synthase Assay
3.7. Immunoblotting Analysis
3.8. Statistical Analysis
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
- Geard, C.R.; Colvett, R.D.; Rohrig, N. On the mechanics of chromosomal aberrations: A study with single and multiple spatially-associated protons. Mutat. Res. 1980, 69, 81–99. [Google Scholar]
- Wu, H.; Durante, M.; George, K.; Yang, T.C. Induction of chromosome aberrations in human cells by charged particles. Radiat. Res. 1997, 48, 102–107. [Google Scholar]
- Skarsgard, L.D.; Wouters, B.G. Substructure in the cell survival response at low radiation dose: Effect of different subpopulations. Int. J. Radiat. Biol. 1997, 71, 737–749. [Google Scholar]
- Ristic-Fira, A.; Nikolic, D.; Petrovic, I.; Ruzdijic, S.; Raffaele, L.; Sabini, M.G.; Cirrone, G.A.; Cuttone, G.; Farruggia, G.; Masotti, L.; et al. The late effects of proton irradiation on cell growth, cell cycle arrest and apoptosis in a human melanoma cell line. J. Exp. Clin. Cancer Res. 2001, 20, 135–143. [Google Scholar]
- Green, L.M.; Tran, D.T.; Murray, D.K.; Rightnar, S.S.; Todd, S.; Nelson, G.A. Response of thyroid follicular cells to gamma irradiation compared to proton irradiation: II. The role of connexin 32. Radiat. Res. 2002, 158, 475–485. [Google Scholar]
- Albi, E.; Cataldi, S.; Rossi, G.; Viola Magni, M.; Toller, M.; Casani, S.; Perrella, G. The nuclear ceramide/diacylglycerol balance depends on the physiological state of thyroid cells and changes during UV-C radiation-induced apoptosis. Arch. Biochem. Biophys. 2008, 478, 52–58. [Google Scholar]
- Ambesi-Impiombato, F.S.; Parks, L.A.; Coon, H.G. Culture of hormone-dependent functional epithelial cells from rat thyroids. Proc. Natl. Acad. Sci. USA 1980, 77, 3455–3459. [Google Scholar]
- De Grassi, A.; Monaco, M.C.; Lisignoli, G.; Belvedere, O.; Toneguzzi, S.; Malangone, W.; Bonora, M.L.; Piacentini, A.; Lavaroni, S.; Scarbolo, M.; et al. Cell cycle synchronization of FRTL5 cells. A physiological model system. J. Exp. Clin. Cancer Res. 1998, 17, 527–532. [Google Scholar]
- Albi, E.; Ambesi-Impiombato, S.; Villani, M.; de Pol, I.; Spelat, R.; Lazzarini, R.; Perrella, G. Thyroid cell growth: Sphingomyelin metabolism as non-invasive marker for cell damage acquired during spaceflight. Astrobiology 2010, 10, 811–820. [Google Scholar]
- Albi, E.; Curcio, F.; Lazzarini, A.; Floridi, A.; Cataldi, S.; Lazzarini, R.; Loreti, E.; Ferri, I.; Ambesi-Impiombato, F.S. A firmer understanding of the effect of hypergravity on thyroid tissue: Cholesterol and thyrotropin receptor. PLoS One 2014, 27, e98250. [Google Scholar]
- Cascianelli, G.; Villani, M.; Tosti, M.; Marini, F.; Bartoccini, E.; Magni, M.V.; Albi, E. Lipid microdomains in cell nucleus. Mol. Biol. Cell 2008, 19, 5289–5295. [Google Scholar]
- Novello, F.; Muchmore, J.H.; Bonora, B.; Capitani, S.; Manzoli, F.A. Effect of phospholipids on the activity of DNA polymerase I from E. coli. Ital. J. Biochem. 1975, 24, 325–334. [Google Scholar]
- Albi, E.; Viola Magni, M.P. Sphingomyelin: A small-big molecule in the nucleus. Recent Res. Dev. Biophys. Biochem. 2006, 661, 211–227. [Google Scholar]
- Ledeen, R.W.; Wu, G. Nuclear sphingolipids: Metabolism and signaling. J. Lipid Res. 2008, 49, 1176–1186. [Google Scholar]
- Albi, E.; Lazzarini, A.; Cataldi, S.; Lazzarini, R.; Floridi, A.; Damaskopoulou, E.; Curcio, F. Nuclear lipid microdomain as place of interaction between sphingomyelin and DNA during Liver regeneration. Int. J. Mol. Sci. 2013, 14, 6529–6541. [Google Scholar]
- Barnes, D.W.H.; Esnouf, M.P.; Stocken, L.A. Some experiments in favour of the cellular hypothesis for the speen curative factor. Adv. Radiobiol. 1957, 211–213. [Google Scholar]
- Matyash, V.; Liebisch, G.; Kurzchalia, T.V.; Shevchenko, A.; Schwudke, D. Lipid extraction by methyl-tert-butyl ether for high-throughput lipidomics. J. Lipid Res. 2008, 49, 1137–1146. [Google Scholar]
- Rabagny, Y.; Herrmann, W.; Geisel, J.; Kirsch, S.H.; Obeid, R. Quantification of plasma phospholipids by ultra performance liquid chromatography tandem mass spectrometry. Anal. Bioanal. Chem. 2011, 401, 891–899. [Google Scholar]
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Albi, E.; Perrella, G.; Lazzarini, A.; Cataldi, S.; Lazzarini, R.; Floridi, A.; Ambesi-Impiombato, F.S.; Curcio, F. Critical Role for the Protons in FRTL-5 Thyroid Cells: Nuclear Sphingomyelinase Induced-Damage. Int. J. Mol. Sci. 2014, 15, 11555-11565. https://doi.org/10.3390/ijms150711555
Albi E, Perrella G, Lazzarini A, Cataldi S, Lazzarini R, Floridi A, Ambesi-Impiombato FS, Curcio F. Critical Role for the Protons in FRTL-5 Thyroid Cells: Nuclear Sphingomyelinase Induced-Damage. International Journal of Molecular Sciences. 2014; 15(7):11555-11565. https://doi.org/10.3390/ijms150711555
Chicago/Turabian StyleAlbi, Elisabetta, Giuseppina Perrella, Andrea Lazzarini, Samuela Cataldi, Remo Lazzarini, Alessandro Floridi, Francesco Saverio Ambesi-Impiombato, and Francesco Curcio. 2014. "Critical Role for the Protons in FRTL-5 Thyroid Cells: Nuclear Sphingomyelinase Induced-Damage" International Journal of Molecular Sciences 15, no. 7: 11555-11565. https://doi.org/10.3390/ijms150711555