Skip to main content

Advertisement

Log in

Effects of photobiomodulation on cellular viability and cancer stem cell phenotype in oral squamous cell carcinoma

  • Original Article
  • Published:
Lasers in Medical Science Aims and scope Submit manuscript

Abstract

Oral squamous cell carcinoma (OSCC) is the most common head and neck malignancy; it has been shown that cancer stem cells (CSC) are present in OSCC and associated with tumor growth, invasion, metastasis, and therapeutic resistance. Photobiomodulation (PBM) is an alternative tool for oncologic treatment adverse effects such as oral mucositis (OM); however, controversy exists regarding the undesirable effects of PBM on tumor or CSC. This study aimed to evaluate in vitro, the effects of PBM, with the same dosimetric parameters as those used in the clinic for OM prevention and treatment, on OSCC cellular viability, as well as PBM’s effect on CSC properties and its phenotype. OSCC cell lines were submitted to single or daily PBM with 3 J/cm2 and 6 J/cm2 and then the cellular viability was evaluated by MTT, NRU (neutral red uptake), and CVS (crystal violet staining). The CSC populations were evaluated by clonogenic formation assay, flow cytometry, and RT-qPCR. The single PBM with the 3 J/cm2 group was associated with increased cellular viability. Daily PBM with 3 J/cm2 and 6 J/cm2 was associated with a significant decrease in cellular viability. Additionally, daily PBM was not able to promote CSC self-renewal or the CD44high/ESAlow and CD44high/ESAhigh cellular phenotypes. Moreover, a decrease in the number of spheres and in the expression of the CSC related gene BMI1 was observed after daily PBM with 6 J/cm2. Daily PBM with 3 J/cm2 and 6 J/cm2 showed an inhibitory effect on cellular viability and was not able to promote the CSC self-renewal or phenotype.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Duprez F, Berwouts D, De Neve W, Bonte K, Boterberg T, Deron P, Huvenne W, Rottery S, Mareel M (2017) Distant metastases in head and neck cancer. Head Neck 39:1733–1743. https://doi.org/10.1002/hed.24687

    Article  PubMed  Google Scholar 

  2. de Vicente JC, Rodríguez-Santamarta T, Rosado P, Peña I, de Villalaín L (2012) Survival after free flap reconstruction in patients with advanced oral squamous cell carcinoma. J Oral Maxillofac Surg 70:453–459. https://doi.org/10.1016/j.joms.2011.02.020

    Article  PubMed  Google Scholar 

  3. Grégoire V, Lefebvre J-L, Licitra L, Felip E, Group ObotEEEGW (2010) Squamous cell carcinoma of the head and neck: EHNS–ESMO–ESTRO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol 21:184–186. https://doi.org/10.1093/annonc/mdq185

    Article  Google Scholar 

  4. Vermorken JB, Specenier P (2010) Optimal treatment for recurrent/metastatic head and neck cancer. Ann Oncol 21:252–261. https://doi.org/10.1093/annonc/mdq453

    Article  Google Scholar 

  5. Nör C, Zhang Z, Warner KA, Bernardi L, Visioli F, Helman JI, Roesler R, Nör JE (2014) Cisplatin induces Bmi-1 and Enhances the stem cell fraction in head and neck cancer. Neoplasia 16:137–138. https://doi.org/10.1593/neo.131744

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Prince ME, Sivanandan R, Kaczorowski A, Wolf GT, Kaplan MJ, Dalerba P, Weissman IL, Clarke MF, Ailles LE (2007) Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. J Proc Natl Acad Sci 104:973–978. https://doi.org/10.1073/pnas.0610117104

    Article  CAS  Google Scholar 

  7. Andrade NPD, Rodrigues MFSD, Rodini CO, Nunes FD (2017) Cancer stem cell, cytokeratins and epithelial to mesenchymal transition markers expression in oral squamous cell carcinoma derived from ortothopic xenoimplantation of CD44high cells. Pathol Res Pract 235:235–244. https://doi.org/10.1016/j.prp.2016.12.009

    Article  CAS  Google Scholar 

  8. Rodrigues MFSD, Xavier FCDA, Andrade NP, Lopes C, Miguita Luiz L, Sedassari BT, Ibarra AMC, López RVM, Kliemann Schmerling C, Moyses RA, Tajara da Silva EE, Nunes FD (2018) Prognostic implications of CD44, NANOG, OCT4, and BMI1 expression in tongue squamous cell carcinoma. Head Neck 40:1759–1773. https://doi.org/10.1002/hed.25158

    Article  PubMed  Google Scholar 

  9. Gemenetzidis E, Gammon L, Biddle A, Emich H, Mackenzie IC (2015) Invasive oral cancer stem cells display resistance to ionising radiation. Oncotarget 22:43964–43977. https://doi.org/10.18632/oncotarget.6268

    Article  Google Scholar 

  10. Wang SJ, Bourguignon LYW (2006) Hyaluronan and the interaction between CD44 and epidermal growth factor receptor in oncogenic signaling and chemotherapy resistance in head and neck cancer. JAMA Otolaryngol Head Neck Surg 132:771–778. https://doi.org/10.1001/archotol.132.7.771

    Article  Google Scholar 

  11. Huang C-F, Xu X-R, Wu T-F, Sun Z-J, Zhang W-F (2014) Correlation of ALDH1, CD44, OCT4 and SOX2 in tongue squamous cell carcinoma and their association with disease progression and prognosis. J Oral Pathol Med 43:492–498. https://doi.org/10.1111/jop.12159

    Article  CAS  PubMed  Google Scholar 

  12. Biddle A, Liang X, Gammon L, Fazil B, Harper LJ, Emich H, Costea DE, Mackenzie IC (2011) Cancer stem cells in squamous cell carcinoma switch between two distinct phenotypes that are preferentially migratory or proliferative. J Cancer Res Ther 71:5317–5326. https://doi.org/10.1158/0008-5472.CAN-11-1059

    Article  CAS  Google Scholar 

  13. De Sanctis V, Bossi P, Sanguineti G, Trippa F, Ferrari D, Bacigalupo A, Ripamonti CI, Buglione M, Pergolizzi S, Langendjik JA, Murphy B, Raber-Durlacher J, Russi EG, Lalla RV (2016) Mucositis in head and neck cancer patients treated with radiotherapy and systemic therapies: literature review and consensus statements. Crit Rev Oncol Hematol 100:147–166. https://doi.org/10.1016/j.critrevonc.2016.01.010

    Article  Google Scholar 

  14. Lalla RV, Bowen J, Barasch A, Elting L, Epstein J, Keefe DM, McGuire DB, Migliorati C, Nicolatou-Galitis O, Peterson DE, Raber-Durlacher JE, Sonis ST, Elad S, Cancer TMGLGotMAoSCi and Oncology. ISoO (2014) MASCC/ISOO clinical practice guidelines for the management of mucositis secondary to cancer therapy. Cancer Metastasis Rev 120:1453–1461. https://doi.org/10.1002/cncr.28592

    Article  Google Scholar 

  15. Antunes HS, Herchenhorn D, Small IA, Araújo CMM, Viégas CMP, de Assis RG, Dias FL, Ferreira CG (2017) Long-term survival of a randomized phase III trial of head and neck cancer patients receiving concurrent chemoradiation therapy with or without low-level laser therapy (LLLT) to prevent oral mucositis. Oral Oncol 71:11–15. https://doi.org/10.1016/j.oraloncology.2017.05.018

    Article  PubMed  Google Scholar 

  16. Zecha JAEM, Raber-Durlacher JE, Nair RG, Epstein JB, Elad S, Hamblin MR, Barasch A, Migliorati CA, Milstein DMJ, Genot M-T, Lansaat L, van der Brink R, Arnabat-Dominguez J, van der Molen L, Jacobi I, van Diessen J, de Lange J, Smeele LE, Schubert MM, Bensadoun R-J (2016) Low-level laser therapy/photobiomodulation in the management of side effects of chemoradiation therapy in head and neck cancer: part 2: proposed applications and treatment protocols. J Support Care Cancer 24:2793–2805. https://doi.org/10.1007/s00520-016-3153-y

    Article  Google Scholar 

  17. Bjordal JM, Bensadoun R-J, Tunèr J, Frigo L, Gjerde K, Lopes-Martins RA (2011) A systematic review with meta-analysis of the effect of low-level laser therapy (LLLT) in cancer therapy-induced oral mucositis. J Support Care Cancer 19:1069–1077. https://doi.org/10.1007/s00520-011-1202-0

    Article  Google Scholar 

  18. Zadik Y, Arany PR, Fregnani ER, Bossi P, Antunes HS, Bensadoun RJ, Gueiros LA, Majorana A, Nair RG, Ranna V, Tissing WJE, Vaddi A, Lubart R, Migliorati CA, Lalla RV, Cheng KKF, Elad S (2019) Systematic review of photobiomodulation for the management of oral mucositis in cancer patients and clinical practice guidelines. Support Care Cancer 27:3969–3983. https://doi.org/10.1007/s00520-019-04890-2

    Article  Google Scholar 

  19. Sonis ST, Hashemi S, Epstein JB, Nair RG, Raber-Durlacher JE (2016) Could the biological robustness of low level laser therapy (photobiomodulation) impact its use in the management of mucositis in head and neck cancer patients. Oral Oncol 54:7–14. https://doi.org/10.1016/j.oraloncology.2016.01.005

    Article  PubMed  Google Scholar 

  20. Sperandio FF, Giudice FS, Corrêa L, Pinto DS Jr, Hamblin MR, de Sousa SCOM (2013) Low-level laser therapy can produce increased aggressiveness of dysplastic and oral cancer cell lines by modulation of Akt/mTOR signaling pathway. J Biophotonics 6:839–847. https://doi.org/10.1002/jbio.201300015

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Bamps M, Dok R, Nuyts S (2018) Low-level laser therapy stimulates proliferation in head and neck squamous cell carcinoma cells. Front Oncol 8:1–6. https://doi.org/10.3389/fonc.2018.00343

    Article  Google Scholar 

  22. Gomes Henriques ÁC, Ginani F, Oliveira RM, Keesen TSL, Galvão Barboza CA, Oliveira Rocha HA, de Castro JFL, Della Coletta R, de Almeida FR (2014) Low-level laser therapy promotes proliferation and invasion of oral squamous cell carcinoma cells. Lasers Med Sci 29:1385–1395. https://doi.org/10.1007/s10103-014-1535-2

    Article  PubMed  Google Scholar 

  23. Dias Schalch T, Porta Santos Fernandes K, Costa-Rodrigues J, Pereira Garcia M, Agnelli Mesquita-Ferrari R, Kalil Bussadori S, Fernandes MH (2016) Photomodulation of the osteoclastogenic potential of oral squamous carcinoma cells. J Biophotonics 9:1136–1147. https://doi.org/10.1002/jbio.201500292

    Article  CAS  PubMed  Google Scholar 

  24. Schartinger VH, Galvan O, Riechelmann H, Dudás J (2012) Differential responses of fibroblasts, non-neoplastic epithelial cells, and oral carcinoma cells to low-level laser therapy. Support Care Cancer 20:523–529. https://doi.org/10.1007/s00520-011-1113-0

    Article  PubMed  Google Scholar 

  25. Schalch TD, Fernandes MH, Destro Rodrigues MFS, Guimarães DM, Nunes FD, Rodrigues JC, Garcia MP, Mesquita Ferrari RA, Bussadori SK, Fernandes KPS (2019) Photobiomodulation is associated with a decrease in cell viability and migration in oral squamous cell carcinoma. Lasers Med Sci 34:629–636. https://doi.org/10.1007/s10103-018-2640-4

    Article  PubMed  Google Scholar 

  26. Ottaviani G, Martinelli V, Rupel K, Caronni N, Naseem A, Zandonà L, Perinetti G, Gobbo M, Di Lenarda R, Bussani R, Benvenuti F, Giacca M, Biasotto M, Zacchigna S (2016) Laser therapy inhibits tumor growth in mice by promoting immune surveillance and vessel normalization. EBioMedicine 11:165–172. https://doi.org/10.1016/j.ebiom.2016.07.028

    Article  PubMed  PubMed Central  Google Scholar 

  27. de Pauli PM, Araújo ALD, Arboleda LPA, Palmier NR, Fonsêca JM, Gomes-Silva W, Madrid-Troconis CC, Silveira FM, Martins MD, Faria KM, Ribeiro ACP, Brandão TB, Lopes MA, Leme AFP, Migliorati CA, Santos-Silva AR (2019) Tumor safety and side effects of photobiomodulation therapy used for prevention and management of cancer treatment toxicities. A systematic review. Oral Oncol 93:21–28. https://doi.org/10.1016/j.oraloncology.2019.04.004

    Article  CAS  Google Scholar 

  28. Mackenzie IC (2004) Growth of malignant oral epithelial stem cells after seeding into organotypical cultures of normal mucosa. J Oral Pathol Med 33:71–78. https://doi.org/10.1111/j.1600-0714.2004.00157.x

    Article  PubMed  Google Scholar 

  29. Silva DFT, Mesquita-Ferrari RA, Fernandes KPS, Raele MP, Wetter NU, Deana AM (2012) Effective transmission of light for media culture, plates and tubes. Photochem Photobiol 88:1211–1216. https://doi.org/10.1111/j.1751-1097.2012.01166.x

    Article  CAS  PubMed  Google Scholar 

  30. Martins WK, Severino D, Souza C, Stolf BS, Baptista MS (2013) Rapid screening of potential autophagic inductor agents using mammalian cell lines. Biotechnol J 8:730–737. https://doi.org/10.1002/biot.201200306

    Article  CAS  PubMed  Google Scholar 

  31. Rodrigues MFSD, Miguita L, De Andrade NP, Heguedusch D, Rodini CO, Moyses RA, Toporcov TN, Gama RR, Tajara EE, Nunes FD (2018) GLI3 knockdown decreases stemness, cell proliferation and invasion in oral squamous cell carcinoma. Int J Oncol 53:2458–2472. https://doi.org/10.3892/ijo.2018.4572

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Djavid GE, Bigdeli B, Goliaei B, Nikoofar A, Hamblin MR (2017) Photobiomodulation leads to enhanced radiosensitivity through induction of apoptosis and autophagy in human cervical cancer cells. J Biophotonics 10:1732–1742. https://doi.org/10.1002/jbio.201700004

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Werneck CE, Pinheiro ALB, Pacheco MTT, Soares CP, Castro JLFD (2005) Laser light is capable of inducing proliferation of carcinoma cells in culture: a spectroscopic in vitro study. Photomed Laser Ther 23:300–303. https://doi.org/10.1089/pho.2005.23.300

    Article  Google Scholar 

  34. Diniz IMA, Souto GR, Freitas IDP, de Arruda JAA, da Silva JM, Silva TA, Mesquita RA (2020) Photobiomodulation enhances cisplatin cytotoxicity in a culture model with oral cell lineages. Photochem Photobiol 96:182–190. https://doi.org/10.1111/php.13152

    Article  CAS  PubMed  Google Scholar 

  35. Pinheiro ALB, Nascimento SC, Vieira ALDB, Brugnera A Jr, Zanin FA, Rolim AB, Silva PSD (2002) Effects of low-level laser therapy on malignant cells: in vitro study. J Clin Laser Med Surg 20:23–26. https://doi.org/10.1089/104454702753474977

    Article  PubMed  Google Scholar 

  36. Al-Watban FAH, Andres BL (2012) Laser biomodulation of normal and neoplastic cells. Lasers Med Sci 27:1039–1043. https://doi.org/10.1007/s10103-011-1040-9

    Article  PubMed  Google Scholar 

  37. Castro JLFD, Pinheiro DALB, Werneck CE, Soares CP (2005) The effect of laser therapy on the proliferation of oral KB carcinoma cells: an in vitro study. Photomed Laser Surg 23:586–589. https://doi.org/10.1089/pho.2005.23.586

    Article  PubMed  Google Scholar 

  38. Powell K, Low P, McDonnell PA, Laakso E-L, Ralph SJ (2010) The effect of laser irradiation on proliferation of human breast carcinoma, melanoma, and immortalized mammary epithelial cells. Photomed Laser Surg 28:115–123. https://doi.org/10.1089/pho.2008.2445

    Article  PubMed  Google Scholar 

  39. Hamblin MR, Demidova TN (2006) Mechanisms of low level light therapy. SPIE. https://doi.org/10.1117/12.646294

  40. Crous A, Heidi A (2016) Low-intensity laser irradiation at 636 nm induces increased viability and proliferation in isolated lung cancer stem cells. Photomed Laser Surg 34:525–532. https://doi.org/10.1089/pho.2015.3979

    Article  PubMed  Google Scholar 

  41. Abrahamse H, Crous A (2016) Biochemical responses of isolated lung CSCs after application of low intensity laser irradiation. Proc. SPIE 9695:96950 J. https://doi.org/10.1117/12.2228902

  42. Huang Y-Y, Chen ACH, Carroll JD and Hamblin MR (2009) Biphasic dose response in low level light therapy. Dose-response: a publication of International Hormesis Society 7:358-383. https://doi.org/10.2203/dose-response.09-027.

  43. Kiro NE, Hamblin MR, Abrahamse H (2017) Photobiomodulation of breast and cervical cancer stem cells using low-intensity laser irradiation. Tumour Biol 39:1010428317706913–1010428317706913. https://doi.org/10.1177/1010428317706913

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Sparmann A, van Lohuizen M (2006) Polycomb silencers control cell fate, development and cancer. Nat Rev Cancer 6:846–856. https://doi.org/10.1038/nrc1991

    Article  CAS  PubMed  Google Scholar 

  45. Hu J, Mirshahidi S, Simental A, Lee SC, De Andrade Filho PA, Peterson NR, Duerksen-Hughes P, Yuan X (2019) Cancer stem cell self-renewal as a therapeutic target in human oral cancer. Oncogene. https://doi.org/10.1038/s41388-019-0800-z

Download references

Funding

This study was financed in part by the Nove de Julho University (UNINOVE) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brasil (CAPES) – Finance Code 001.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Maria Fernanda Setúbal Destro Rodrigues.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

ESM 1

(DOCX 14 kb)

ESM 2

(JPG 44 kb)

ESM 3

(JPG 211 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ibarra, A.M.C., Garcia, M.P., Ferreira, M. et al. Effects of photobiomodulation on cellular viability and cancer stem cell phenotype in oral squamous cell carcinoma. Lasers Med Sci 36, 681–690 (2021). https://doi.org/10.1007/s10103-020-03131-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10103-020-03131-x

Keywords

Navigation