Skip to main content
Top
Published in: Lasers in Medical Science 1/2018

01-01-2018 | Original Article

cDNA microarray analysis of human keratinocytes cells of patients submitted to chemoradiotherapy and oral photobiomodulation therapy: pilot study

Authors: Heliton S. Antunes, Gabriel Wajnberg, Marcos B. Pinho, Natasha Andressa Nogueira Jorge, Joyce Luana Melo de Moraes, Claudio Gustavo Stefanoff, Daniel Herchenhorn, Carlos M. M. Araújo, Celia Maria Pais Viégas, Mariana P. Rampini, Fernando L. Dias, Patricia Savio de Araujo-Souza, Fabio Passetti, Carlos G. Ferreira

Published in: Lasers in Medical Science | Issue 1/2018

Login to get access

Abstract

Oral mucositis is an acute toxicity that occurs in patients submitted to chemoradiotherapy to treat head and neck squamous cell carcinoma. In this study, we evaluated differences in gene expression in the keratinocytes of the oral mucosa of patients treated with photobiomodulation therapy and tried to associate the molecular mechanisms with clinical findings. From June 2009 to December 2010, 27 patients were included in a randomized double-blind pilot study. Buccal smears from 13 patients were obtained at days 1 and 10 of chemoradiotherapy, and overall gene expression of samples from both dates were analyzed by complementary DNA (cDNA) microarray. In addition, samples from other 14 patients were also collected at D1 and D10 of chemoradiotherapy for subsequent validation of cDNA microarray findings by qPCR. The expression array analysis identified 105 upregulated and 60 downregulated genes in our post-treatment samples when compared with controls. Among the upregulated genes with the highest fold change, it was interesting to observe the presence of genes related to keratinocyte differentiation. Among downregulated genes were observed genes related to cytotoxicity and immune response. The results indicate that genes known to be induced during differentiation of human epidermal keratinocytes were upregulated while genes associated with cytotoxicity and immune response were downregulated in the laser group. These results support previous clinical findings indicating that the lower incidence of oral mucositis associated with photobiomodulation therapy might be correlated to the activation of genes involved in keratinocyte differentiation.
Appendix
Available only for authorised users
Literature
1.
go back to reference Adelstein DJ, Li Y, Adams GL et al (2003) An intergroup phase III comparison of standard radiation therapy and two schedules of concurrent chemoradiotherapy in patients with unresectable squamous cell head and neck cancer. J Clin Oncol 21(1):92–98CrossRefPubMed Adelstein DJ, Li Y, Adams GL et al (2003) An intergroup phase III comparison of standard radiation therapy and two schedules of concurrent chemoradiotherapy in patients with unresectable squamous cell head and neck cancer. J Clin Oncol 21(1):92–98CrossRefPubMed
2.
go back to reference Trotti A, Bellm LA, Epstein JB et al (2003) Mucositis incidence, severity and associated outcomes in patients with head and neck cancer receiving radiotherapy with or without chemotherapy: a systematic literature review. Radiother Oncol 66(3):253–262CrossRefPubMed Trotti A, Bellm LA, Epstein JB et al (2003) Mucositis incidence, severity and associated outcomes in patients with head and neck cancer receiving radiotherapy with or without chemotherapy: a systematic literature review. Radiother Oncol 66(3):253–262CrossRefPubMed
3.
go back to reference Sonis ST (2004) The pathobiology of mucositis. Nat Rev Cancer 4:277–284 Sonis ST (2004) The pathobiology of mucositis. Nat Rev Cancer 4:277–284
4.
go back to reference Sonis S, Peterson R, Edwards L (2003) Defining mechanisms of action of interleukin-11 on the progression of radiation-induced mucositis in hamsters. Oral Oncol 6:393–381 Sonis S, Peterson R, Edwards L (2003) Defining mechanisms of action of interleukin-11 on the progression of radiation-induced mucositis in hamsters. Oral Oncol 6:393–381
5.
go back to reference Xanthinaki A, Nicolatou-Galitis O, Athanassiadou P et al (2008) Apoptotic and inflammation markers in oral mucositis in head and neck cancer patients receiving radiotherapy: preliminary report. Support Care Cancer 16(9):1025–1033CrossRefPubMed Xanthinaki A, Nicolatou-Galitis O, Athanassiadou P et al (2008) Apoptotic and inflammation markers in oral mucositis in head and neck cancer patients receiving radiotherapy: preliminary report. Support Care Cancer 16(9):1025–1033CrossRefPubMed
6.
go back to reference Barasch A, Peterson DE, Tanzer JM et al (1995) Helium-neon laser effects on conditioning-induced oral mucositis in bone marrow transplantation patients. Cancer 76:2550–2556CrossRefPubMed Barasch A, Peterson DE, Tanzer JM et al (1995) Helium-neon laser effects on conditioning-induced oral mucositis in bone marrow transplantation patients. Cancer 76:2550–2556CrossRefPubMed
7.
go back to reference Cowen D, Tardieu C, Schubert M et al (1997) Low energy helium–neon laser in the prevention of oral mucositis in patients undergoing bone marrow transplantation: results of a double blind randomized trial. Int. J. Radiation Oncology Biol. Phys 38:697–703CrossRef Cowen D, Tardieu C, Schubert M et al (1997) Low energy helium–neon laser in the prevention of oral mucositis in patients undergoing bone marrow transplantation: results of a double blind randomized trial. Int. J. Radiation Oncology Biol. Phys 38:697–703CrossRef
8.
go back to reference Antunes HS, Azevedo AM, Bouzas LFS et al (2007) Low power laser in the prevention of induced oral mucositis in bone marrow transplantation patients: a randomized trial. Blood 109:2250–2255CrossRefPubMed Antunes HS, Azevedo AM, Bouzas LFS et al (2007) Low power laser in the prevention of induced oral mucositis in bone marrow transplantation patients: a randomized trial. Blood 109:2250–2255CrossRefPubMed
9.
go back to reference Schubert MM, Eduardo FP, Guthrie KA et al (2007) A phase III randomized double-blind placebo-controlled clinical trial to determine the efficacy of low level laser therapy for the prevention of oral mucositis in patients undergoing hematopoietic cell transplantation. Support Care Cancer 15:1145–1154CrossRefPubMed Schubert MM, Eduardo FP, Guthrie KA et al (2007) A phase III randomized double-blind placebo-controlled clinical trial to determine the efficacy of low level laser therapy for the prevention of oral mucositis in patients undergoing hematopoietic cell transplantation. Support Care Cancer 15:1145–1154CrossRefPubMed
10.
go back to reference Silva GBL, Mendonça EF, Bariani C, Antunes HS, Silva MAG (2011) The prevention of induced oral mucositis with low-level laser therapy in bone marrow transplantation patients: a randomized clinical trial. Photomed Laser Surg 29:27–31CrossRefPubMed Silva GBL, Mendonça EF, Bariani C, Antunes HS, Silva MAG (2011) The prevention of induced oral mucositis with low-level laser therapy in bone marrow transplantation patients: a randomized clinical trial. Photomed Laser Surg 29:27–31CrossRefPubMed
11.
go back to reference Bensadoun RJ, Franquin JC, Ciais G et al (1999) Low-energy he/ne laser in the prevention of radiation-induced mucositis: a multicenter phase III randomized study in patients with head and neck cancer. Suport Care Cancer 7:244–252CrossRef Bensadoun RJ, Franquin JC, Ciais G et al (1999) Low-energy he/ne laser in the prevention of radiation-induced mucositis: a multicenter phase III randomized study in patients with head and neck cancer. Suport Care Cancer 7:244–252CrossRef
12.
go back to reference Maiya GA, Sagar MS, Fernandes D (2006) Effect of low helium neon (He-Ne) laser therapy in the prevention and treatment of radiation-induced mucositis in head and neck cancer patients. Indian J Med Res 124:399–402 Maiya GA, Sagar MS, Fernandes D (2006) Effect of low helium neon (He-Ne) laser therapy in the prevention and treatment of radiation-induced mucositis in head and neck cancer patients. Indian J Med Res 124:399–402
13.
go back to reference Arora H, Pai KM, Maiya GA, Vidyasagar MS, Rajeev A (2008) Efficacy of He-Ne laser in the prevention and treatment of radiotherapy-induced oral mucositis in oral cancer patients. Oral Surg Oral Med Oral Pathol Oral Radiol Oral Endod 105:180–186CrossRef Arora H, Pai KM, Maiya GA, Vidyasagar MS, Rajeev A (2008) Efficacy of He-Ne laser in the prevention and treatment of radiotherapy-induced oral mucositis in oral cancer patients. Oral Surg Oral Med Oral Pathol Oral Radiol Oral Endod 105:180–186CrossRef
14.
go back to reference Gouveia de Lima A, Villar RG, Castro JRG et al (2012) Oral mucositis prevention by low-level laser therapy in head and neck cancer patients undergoing concurrent chemoradiotherapy: a phase III randomized study. Int J Radiation Oncology Biol Phys 82:270–275CrossRef Gouveia de Lima A, Villar RG, Castro JRG et al (2012) Oral mucositis prevention by low-level laser therapy in head and neck cancer patients undergoing concurrent chemoradiotherapy: a phase III randomized study. Int J Radiation Oncology Biol Phys 82:270–275CrossRef
15.
go back to reference Carvalho PA, Jaguar GC, Pellizzon AC et al (2011) Evaluation of low-level laser therapy in the prevention and treatment of radiation-induced mucositis: a double-blind randomized studyin head and neck cancer patients. Oral Oncol 47:1176–1181CrossRefPubMed Carvalho PA, Jaguar GC, Pellizzon AC et al (2011) Evaluation of low-level laser therapy in the prevention and treatment of radiation-induced mucositis: a double-blind randomized studyin head and neck cancer patients. Oral Oncol 47:1176–1181CrossRefPubMed
16.
go back to reference Gautam AP, Fernandes DJ, Vidyasagar MS, Maiya AG (2012) Low level helium neon laser therapy for chemoradiotherapy induced oral mucositis in oral cancer patients—a randomized controlled trial. Oral Oncol 48(9):893–897CrossRefPubMed Gautam AP, Fernandes DJ, Vidyasagar MS, Maiya AG (2012) Low level helium neon laser therapy for chemoradiotherapy induced oral mucositis in oral cancer patients—a randomized controlled trial. Oral Oncol 48(9):893–897CrossRefPubMed
17.
go back to reference Gautam AP, Fernandes DJ, Vidyasagar MS, Maiya AG, Vadhiraja BM (2012) Low level laser therapy for concurrent chemoradiotherapy induced oral mucositis in head and neck cancer patients. A triple blinded randomized controlled trial. Radiother Oncol 104(3):349CrossRefPubMed Gautam AP, Fernandes DJ, Vidyasagar MS, Maiya AG, Vadhiraja BM (2012) Low level laser therapy for concurrent chemoradiotherapy induced oral mucositis in head and neck cancer patients. A triple blinded randomized controlled trial. Radiother Oncol 104(3):349CrossRefPubMed
18.
go back to reference Antunes HS, Herchenhorn D, Small I et al (2013) Phase III trial of low-level laser therapy to prevent oral mucositis in head and neck cancer patients treated with concurrent chemoradiation. Radiother Oncol 109:297–302CrossRefPubMed Antunes HS, Herchenhorn D, Small I et al (2013) Phase III trial of low-level laser therapy to prevent oral mucositis in head and neck cancer patients treated with concurrent chemoradiation. Radiother Oncol 109:297–302CrossRefPubMed
19.
go back to reference Yu H-S, Chang K-L, Yu C-L (1996) Low-energy hellion neon laser irradiation stimulates interleukin-1alfa and interleukin-8 release from cultured human keratinocytes. The Journal of Investigate Dermatology 107(4):593–596CrossRef Yu H-S, Chang K-L, Yu C-L (1996) Low-energy hellion neon laser irradiation stimulates interleukin-1alfa and interleukin-8 release from cultured human keratinocytes. The Journal of Investigate Dermatology 107(4):593–596CrossRef
21.
go back to reference Karu T (1999) Primary and secondary mechanisms of action of visible-to-near irradiation on cells. J Photochem Photobiol B 49(1):1–17CrossRefPubMed Karu T (1999) Primary and secondary mechanisms of action of visible-to-near irradiation on cells. J Photochem Photobiol B 49(1):1–17CrossRefPubMed
22.
go back to reference Karu T, Afanasyeva NI, Kolyakov SF, Pyatibrat LV, Welser L (2001) Changes in absorvance of monolayer of living cells induced by laser radiation at 633,670, and 820 nm. Quantum Eletronic 7(6):982–988 Karu T, Afanasyeva NI, Kolyakov SF, Pyatibrat LV, Welser L (2001) Changes in absorvance of monolayer of living cells induced by laser radiation at 633,670, and 820 nm. Quantum Eletronic 7(6):982–988
23.
go back to reference Karu TI, Pyatibrat LV, Afanasyeva NI (2004) A novel mitochondrial signaling pathway activated by visible-to-near infrared radiation. Photochem Photobiol 80(2):366–372CrossRefPubMed Karu TI, Pyatibrat LV, Afanasyeva NI (2004) A novel mitochondrial signaling pathway activated by visible-to-near infrared radiation. Photochem Photobiol 80(2):366–372CrossRefPubMed
24.
go back to reference Karu TI, Pyatibrat LV, Kolyakov SF, Afanasyeva NI (2008) Absorption measurements of cell monolayers relevant to mechanisms of laser phototherapy: reduction or oxidation of cytochrome c oxidase under laser radiation at 632.8 nm. Photomed Laser Surg 26(6):593–599CrossRefPubMed Karu TI, Pyatibrat LV, Kolyakov SF, Afanasyeva NI (2008) Absorption measurements of cell monolayers relevant to mechanisms of laser phototherapy: reduction or oxidation of cytochrome c oxidase under laser radiation at 632.8 nm. Photomed Laser Surg 26(6):593–599CrossRefPubMed
25.
go back to reference Alexandratou E, Yova D, Handris P, Kletsas D, Loukas S (2002) Human fibroblast alterations induced by low power laser irradiation at the single cell level using confocal microscopy. Photochem PhotobiolSci 1(8):547–552CrossRef Alexandratou E, Yova D, Handris P, Kletsas D, Loukas S (2002) Human fibroblast alterations induced by low power laser irradiation at the single cell level using confocal microscopy. Photochem PhotobiolSci 1(8):547–552CrossRef
26.
go back to reference Zhang Y, Song S, Fong CC, Tsang CH, Yang Z, Yang M (2003) cDNA microarray analisis of gene expression profiles in human fibroblast cells irradiated with red light. J Invest Dermatol 120(5):849–857CrossRefPubMed Zhang Y, Song S, Fong CC, Tsang CH, Yang Z, Yang M (2003) cDNA microarray analisis of gene expression profiles in human fibroblast cells irradiated with red light. J Invest Dermatol 120(5):849–857CrossRefPubMed
27.
go back to reference Safavi SM, Kazemi B, Esmaeili M, Fallah A, Modarresi A, Mir M (2008) Effects of low-level He–Ne laser irradiation on the gene expression of IL-1β, TNF-α, IFN-γ, TGF-β, bFGF, and PDGF in rat’s gingiva. Lasers Med Sci 23(3):331–335CrossRefPubMed Safavi SM, Kazemi B, Esmaeili M, Fallah A, Modarresi A, Mir M (2008) Effects of low-level He–Ne laser irradiation on the gene expression of IL-1β, TNF-α, IFN-γ, TGF-β, bFGF, and PDGF in rat’s gingiva. Lasers Med Sci 23(3):331–335CrossRefPubMed
28.
go back to reference Saygun I, Karacay S, Serdar M et al (2008) Effects of laser irradiation on the release of basic fibroblast growth factor (bFGF), insulin like growth factor-1 (IGF-1), and receptor of IGF-1 (IGFBP3) from gingival fibroblasts. Lasers Med Sci 23(2):211–215CrossRefPubMed Saygun I, Karacay S, Serdar M et al (2008) Effects of laser irradiation on the release of basic fibroblast growth factor (bFGF), insulin like growth factor-1 (IGF-1), and receptor of IGF-1 (IGFBP3) from gingival fibroblasts. Lasers Med Sci 23(2):211–215CrossRefPubMed
29.
go back to reference Mvula B, Moore TJ, Abrahamse H (2010) Effect of low-level laser irradiation and epidermal growthfactor on adult human adipose-derived stem cells. Lasers Med Sci 25(1):33–39CrossRefPubMed Mvula B, Moore TJ, Abrahamse H (2010) Effect of low-level laser irradiation and epidermal growthfactor on adult human adipose-derived stem cells. Lasers Med Sci 25(1):33–39CrossRefPubMed
30.
go back to reference WHO (1979) Handbook for reporting results of cancer treatment. WHO Offset Publication No. 48. World Health Organization, Geneva WHO (1979) Handbook for reporting results of cancer treatment. WHO Offset Publication No. 48. World Health Organization, Geneva
31.
go back to reference Sonis ST, Eilers JP, Epstein JB et al (1999) Validation of a new scoring system for the assessment of clinical trial research of oral mucositis induced by radiation or chemotherapy. Cancer 85:2103–2113CrossRefPubMed Sonis ST, Eilers JP, Epstein JB et al (1999) Validation of a new scoring system for the assessment of clinical trial research of oral mucositis induced by radiation or chemotherapy. Cancer 85:2103–2113CrossRefPubMed
33.
go back to reference Reimers M, Carey VJ (2006) Bioconductor: an open source framework for bioinformatics and computational biology. Methods Enzymol 411:119–134CrossRefPubMed Reimers M, Carey VJ (2006) Bioconductor: an open source framework for bioinformatics and computational biology. Methods Enzymol 411:119–134CrossRefPubMed
34.
go back to reference Zhang J (2017). tkWidgets: R based tk widgets. R package Zhang J (2017). tkWidgets: R based tk widgets. R package
35.
go back to reference Irizarry RA, Hobbs B, Collin F et al (2003) Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 4(2):249–264CrossRefPubMed Irizarry RA, Hobbs B, Collin F et al (2003) Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 4(2):249–264CrossRefPubMed
36.
go back to reference Gautier L, Cope L, Bolstad BM, Irizarry RA (2004) Affy-analysis of Affymetrix GeneChip data at the probe level. Bioinformatics 20(3):307–315CrossRefPubMed Gautier L, Cope L, Bolstad BM, Irizarry RA (2004) Affy-analysis of Affymetrix GeneChip data at the probe level. Bioinformatics 20(3):307–315CrossRefPubMed
37.
go back to reference Gentleman R, Carey V, Huber W and Hahne F. Genefilter: genefilter: methods for filtering genes from high-throughput experiments. R package version 1.50.0 Gentleman R, Carey V, Huber W and Hahne F. Genefilter: genefilter: methods for filtering genes from high-throughput experiments. R package version 1.50.0
38.
go back to reference Smyth GK (2005) Limma: linear models for microarray data. In: Gentleman R, Carey V, Dudoit S, Irizarry R, Huber W (eds) Bioinformatics and computational biology solutions using R and bioconductor. Springer, New York, pp 397–420CrossRef Smyth GK (2005) Limma: linear models for microarray data. In: Gentleman R, Carey V, Dudoit S, Irizarry R, Huber W (eds) Bioinformatics and computational biology solutions using R and bioconductor. Springer, New York, pp 397–420CrossRef
39.
go back to reference Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing (PDF). J R Stat Soc Ser B 57(1):289–300 Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing (PDF). J R Stat Soc Ser B 57(1):289–300
41.
42.
go back to reference Morse DL, Carroll D, Weberg L et al (2005) Determining suitable internal standards for mRNA quantification of increasing cancer progression in human breast cells by real-time reverse transcriptase polymerase chain reaction. Anal Biochem 342(1):69–77CrossRefPubMed Morse DL, Carroll D, Weberg L et al (2005) Determining suitable internal standards for mRNA quantification of increasing cancer progression in human breast cells by real-time reverse transcriptase polymerase chain reaction. Anal Biochem 342(1):69–77CrossRefPubMed
43.
go back to reference Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C (T)) method. Methods 25(4):402–408CrossRefPubMed Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C (T)) method. Methods 25(4):402–408CrossRefPubMed
44.
go back to reference Tsai CA, Wang SJ, Chen DT, Chen JJ (2005) Sample size for gene expression microarray experiments. Bioinformatics 21(8):1502–1508CrossRefPubMed Tsai CA, Wang SJ, Chen DT, Chen JJ (2005) Sample size for gene expression microarray experiments. Bioinformatics 21(8):1502–1508CrossRefPubMed
45.
go back to reference Gibbs S, Fijneman R, Wiegant J et al (1993) Molecular characterization and evolution of the SPRR family of keratinocyte differentiation markers encoding small proline-rich proteins. Genomics 16(3):630–637 [Internet]. Accessed 13 Jan 2015. Available from http://www.ncbi.nlm.nih.gov/pubmed/8325635 Gibbs S, Fijneman R, Wiegant J et al (1993) Molecular characterization and evolution of the SPRR family of keratinocyte differentiation markers encoding small proline-rich proteins. Genomics 16(3):630–637 [Internet]. Accessed 13 Jan 2015. Available from http://​www.​ncbi.​nlm.​nih.​gov/​pubmed/​8325635
46.
go back to reference Antunes HS, Ferreira EMS, Mattos VD, Pinheiro CT, Ferreira CG (2008) The impact of low power laser in the treatment of conditioning-induced oral mucositis: a report of 11 clinical cases and their review. Med Oral Patol Oral Cir Bucal 13(3):E189–E192PubMed Antunes HS, Ferreira EMS, Mattos VD, Pinheiro CT, Ferreira CG (2008) The impact of low power laser in the treatment of conditioning-induced oral mucositis: a report of 11 clinical cases and their review. Med Oral Patol Oral Cir Bucal 13(3):E189–E192PubMed
Metadata
Title
cDNA microarray analysis of human keratinocytes cells of patients submitted to chemoradiotherapy and oral photobiomodulation therapy: pilot study
Authors
Heliton S. Antunes
Gabriel Wajnberg
Marcos B. Pinho
Natasha Andressa Nogueira Jorge
Joyce Luana Melo de Moraes
Claudio Gustavo Stefanoff
Daniel Herchenhorn
Carlos M. M. Araújo
Celia Maria Pais Viégas
Mariana P. Rampini
Fernando L. Dias
Patricia Savio de Araujo-Souza
Fabio Passetti
Carlos G. Ferreira
Publication date
01-01-2018
Publisher
Springer London
Published in
Lasers in Medical Science / Issue 1/2018
Print ISSN: 0268-8921
Electronic ISSN: 1435-604X
DOI
https://doi.org/10.1007/s10103-017-2313-8

Other articles of this Issue 1/2018

Lasers in Medical Science 1/2018 Go to the issue