UVC Radiation-induced Effect on Human Primary Thyroid Cell Proliferation and HLA-DR Expression

Abstract

The aim of this study was to examine how UVC irradiation will affect normal human thyroid cell proliferation and HLA-DR expression. Primary human thyroid cells were exposed to UVC (254 nm wavelength) irradiation. In some experiments 0.5 mM buthionine sulfoximine (BSO) was added. Apoptosis was detected measuring annexin V, proteins involved in apoptotic process (p53, Bax, Bcl-2, caspase 3, and 9) by immunoblot analysis and HLA-DR expression by FACS. UVC induced a cell cycle arrest in G0/G1 phase in the first 24 h, accumulation of cells in the S phase 72 h after treatment, and an increase of apoptotic cells. BSO pretreatment showed an earlier appearance and a higher percentage of apoptosis. p53, caspase 3 and 9 were increased, while Bax and Bcl-2 were decreased. We also observed a transient significant increase in HLA-DR expression. UVC inhibited cell proliferation and induced apoptosis in normal human primary thyroid cells. An inhibitor of glutathione synthesis induced an earlier appearance and higher percentage of apoptosis suggesting that oxidative stress may play a role. Apoptotis involved components of the intrinsic mitochondrial pathway. A transient increase in HLA-DR expression after UVC irradiation could play a role in inducing AITD.

UVC irradiation - human primary thyroid cells - cell cycle - apoptosis - HLA-DR

References

• 1 Godar DE, Lucas AD. Spectral dependence of UV-induced immediate and delayed apoptosis: The role of membrane and DNA damage.  Photochem Photobiol. 1995;  62 108-113
• 2 Young AR. The sunburn cell.  Photodermatol. 1987;  4 127-134
• 3 Allan LA, Fried M. p53-dependent apoptosis or growth arrest induced by different forms of radiation in U20S cells: p21WAF1/CIP1 repression in UV induced apoptosis.  Oncogene. 1999;  18 5403-5412
• 4 Radziszewska E, Piwocka K, Skierski J, Sikora E. UVC-induced cell death of IL-2-dependent human lymphocytes.  Cell Biol Int. 1999;  23 97-103
• 5 Kimura C, Zhao QL, Kondo T, Amatsu M, Fujiwara Y. Mechanism of UV-induced apoptosis in human leukemia cells: Roles of Ca²⁺/Mg²⁺-dependent endonuclease, caspase-3, and stress-activated protein kinases.  Exp Cell Res. 1998;  239 411-422
• 6 Shaulian E, Schreiber M, Piu F, Beeche M, Wagner EF, Karin M. The mammalian UV response: c-Jun induction is required for exit from p53-imposed growth arrest.  Cell. 2000;  103 897-907
• 7 Sarasin A, Bounacer A, Lepage F, Schlumberger M, Guillermo Suarez H. Mechanisms of mutagenesis in mammalian cells. Application to human thyroid tumors.  CR Acad Sci III. 1999;  322 143-149
• 8 Lautier D, Luscher P, Tyrrel RM. Endogenous glutathione levels modulate both constitutive and UV-A radiation/hydrogen peroxide inducible expression of the human heme oxigenase gene.  Carcinogenesis. 1992;  13 227-232
• 9 Devary Y, Gottlieb RA, Smeal T, Karin M. The mammalian ultraviolet response is triggered by activation of src thyrosine kinases.  Cell. 1992;  71 1081-1091
• 10 Hartwell LH, Kastani MB. Cell cycle control and cancer.  Science. 1994;  266 1821-1828
• 11 Mach B, Steimle V, Martinez-Soria E, Reith W. Regulation of MHC class II genes: lessons from a disease.  Ann Rev Immunol. 1996;  14 301-331
• 12 Wu Z, Biro PA, Mirakian R, Hammond L, Curcio F, Ambesi-Impiombato FS. HLA-DMB expression by thyrocytes: indication of the antigen-processing and possible presenting capability of thyroid cells.  Clin Exp Immunol. 1999;  116 62-69
• 13 Czirjak L, Danko K, Gaulton GN, Stadecker MJ. Thyroid-derived epithelial cells acquire alloantigen-presenting capabilities following X-irradiation and class II antigen induction.  Eur J Immunol. 1990;  20 2597-2601
• 14 Botazzo GF, Pujol-Borrell R, Hanafusa T, Feldmann M. Role of aberrant HLA-DR expression and antigen presentation in induction of endocrine autoimmunity.  Lancet. 1983;  2 ((8359)) 1115-1119
• 15 Del Terra E, Francesconi A, Donnini D, Curcio F, Ambesi-Impiombato FS. Thyrotropin effects on UV-dependent apoptosis in FRTL-5 cells.  Thyroid. 2003;  13 747-753
• 16 Lonigro R, Donnini D, Fabbro D, Perrella G, Damante G, Ambesi-Impiombato FS, Curcio F. Thyroid-specific gene expression is differentially influenced by intracellular glutathione level in FRTL-5 cells.  Endocrinology. 2000;  141 901-909
• 17 Sato H, Abe Y, Noguchi M, Kurosawa K, Sakai H. Inhibitory effect of thyrotropic hormone on apoptosis induced by actinomycin D in a functioning rat thyroid cell line.  Endocr J. 1999;  46 309-315
• 18 Curcio F, Ambesi-Impiombato FS, Perrella G, Coon HG. Long-term culture and functional characterization of follicular cells from adult normal human thyroids.  Proc Natl Acad Sci USA. 1994;  91 9004-9008
• 19 Kim H, Lee TH, Park ES, Suh JM, Park SJ, Chung HK, Kwon OY, Kim YK, Ro HK, Shong M. Role of peroxiredoxins in regulating intracellular hydrogen peroxide and hydrogen peroxide-induced apoptosis in thyroid cells.  J Biol Chem. 2000;  275 18266-18270
• 20 McKay BC, Becerril C, Spronck JC, Ljungman M. Ultraviolet light-induced apoptosis is associated with S-phase in primary human fibroblasts.  DNA Repair. 2002;  1 811-820
• 21 Kostic I, Toffoletto B, Fontanini E, Moretti M, Cesseli D, Beltrami CA, Ambesi Impiombato FS, Curcio F. Influence of iodide excess and IFN-γ on human primary thyroid cell proliferation, thyroglobulin secretion, and ICAM-1 and HLA DR expression.  Thyroid. 2009;  19 283-291
• 22 Reinke V, Lozano G. Differential activation of p53 targets in cells treated with ultraviolet radiation that undergo both apoptosis and growth arrest.  Radiat Res. 1997;  148 115-122
• 23 Namba H, Hara T, Tukazaki T, Migita K, Ishikawa N, Ito K, Nagataki S, Yamashita S. Radiation-induced G1 arrest is selectively mediated by the p53-WAF1/Cip1 pathway in human thyroid cells.  Cancer Res. 1995;  55 2075-2080
• 24 McKay BC, Ljungman M, Rainbow AJ. Persistant DNA damage induced by ultraviolet light inhibits p21waf1 and bax expression: implications for DNA repair, UV sensitivity and the induction of apoptosis.  Oncogene. 1998;  17 545-555
• 25 Chang D, Chen F, Zhang F, McKay BC, Ljungman M. Dose-dependent affects of DNA-damaging agents on p53-mediated cell cycle arrest.  Cell Growth and Differentiation. 1999;  10 155-162
• 26 Polyak K, Waldman T, He TC, Kinzler KW, Vogelstein B. Genetic determinants of p53-induced apoptosis and growth arrest.  Genes Dev. 1996;  10 1945-1952
• 27 Kaneko YS, Ikeda K, Nakanishi M. Phorbol ester inhibits DNA damage-induced apoptosis in U937 cells through activation of protein kinase C.  Life Sci. 1999;  65 2251-2258
• 28 Montani V, Taniguchi SI, Shong M, Suzuki K, Ohmori M, Giuliani C, Napolitano G, Saji M, Fiorentino B, Reimold AM, Ting JP, Kohn LD, Singer DS. Major histocompatibility class II HLA DR gene expression in thyrocytes: counter regulation by the class II transactivator and the thyroid Y box protein.  Endocrinology. 1998;  139 280-289
• 29 Otten LA, Steimle V, Bontron S, Mach B. Quantitative control of MHC class II expression by the transactivator CIITA.  Eur J Immunol. 1998;  28 473-478
• 30 Domberg J, Chao L, Papewalis C, Pfleger C, Xu K, Willenberg HS, Hermsen D, Scherbaum WA, Schloot NC, Schott M. Circulating chemokines in patients with autoimmune thyroid diseases.  Horm Metab Res. 2008;  40 416-421
• 31 Liu C, Papewalis C, Domberg J, Scherbaum WA, Schott M. Chemokines and autoimmune thyroid diseases.  Horm Metab Res. 2008;  40 361-368
• 32 McKay BC, Ljungman M. Role for p53 in the recovery of transcription and protection against apoptosis induced by ultraviolet light.  Neoplasia. 1999;  1 276-284
• 33 Wang SH, Koenig RJ, Giordano TJ, Myc A, Thompson HW, Baker JR Jr. 1Alpha, 25-dihydroxyvitamin D3 up-regulates Bcl-2 expression and protects normal human thyrocytes from programmed cell death.  Endocrinology. 1999;  140 1649-1656
• 34 Adams JM, Cory S. Life-or-death decisions by the Bcl-2 protein family.  Trends Biochem Sci. 2001;  26 61-66
• 35 Voehringer DW, Meyn RE. Redox aspects of Bcl-2 function.  Antioxid Redox Signal. 2000;  2 537-550
• 36 Meister A. Glutathione metabolism.  Methods Enzymol. 1995;  251 3-7
• 37 Dunkern TR, Fritz G, Kaina B. Ultraviolet light-induced DNA damage triggers apoptosis in nucleotide excision repair-deficient cells via Bcl-2 decline and caspase-3/-8 activation.  Oncogene. 2001;  20 6026-6038
• 38 Gross A, McDonnell JM, Korsmeyer SJ. BCL-2 family members and the mitochondria in apoptosis.  Genes Dev. 1999;  13 1899-1911
• 39 Tomicic MT, Christmann M, Kaina B. Apoptosis in UV-C light irradiated p53 wild-type, apaf-1 and p53 knockout mouse embryonic fibroblasts: interplay of receptor and mitochondrial pathway.  Apoptosis. 2005;  10 1295-1304
• 40 Kim E, Ma WL, Lin DL, Inui S, Chen YL, Chang C. TR4 orphan receptor functions as an apoptosis modulator via regulation of Bcl-2 gene expression.  Biochem Biophys Res Commun. 2007;  361 323-328

Correspondence