In vitro and in silico validation of CA3 and FHL1 downregulation in oral cancer.

BACKGROUND: Aberrant methylation is a frequent event in oral cancer. METHODS: In order to better characterize these alterations, a search for genes downregulated by aberrant methylation in oral squamous cell carcinoma (OSCC) was conducted through the mining of ORESTES dataset. Findings were further validated in OSCC cell lines and patients' samples and confirmed using TCGA data. Differentially expressed genes were identified in ORESTES libraries and validated in vitro using RT-PCR in HNSCC cell-lines and OSCC tumor samples. Further confirmation of these results was performed using mRNA expression and methylation data from The Cancer Genome Atlas (TCGA) data. RESULTS: From the set of genes selected for validation, CA3 and FHL1 were downregulated in 60% (12/20) and 75% (15/20) of OSCC samples, respectively, and in HNSCC cell lines. The treatment of cell lines JHU-13 and FaDu with the demethylating agent 5'-aza-dC was efficient in restoring CA3 and FHL1 expression. TCGA expression and methylation data on OSCC confirms the downregulation of these genes in OSCC samples and also suggests that expression of CA3 and FHL1 is probably regulated by methylation. The downregulation of CA3 and FHL1 observed in silico was validated in HNSCC cell lines and OSCC samples, showing the feasibility of integrating different datasets to select differentially expressed genes in silico. CONCLUSIONS: These results showed that the downregulation of CA3 and FHL1 data observed in the ORESTES libraries was validated in HNSCC cell lines and OSCC samples and in a large cohort of samples from the TCGA database. Moreover, it suggests that expression of CA3 and FHL1 could probably be regulated by methylation having an important role the oral carcinogenesis.

Oral Mucositis in Cancer and Potential Use of Omega-3 Free Fatty Acids in Its Management: A Review.

Oral mucositis (OM) is a painful condition caused by chemotherapeutic or radiotherapeutic cancer treatments, occurring in patients with different tumour characteristics and locations. OM greatly impacts a patient's quality of life and cancer recovery. Current OM management strategies are not providing sufficient prevention and treatment; new approaches to injury management are needed. Studies on the benefit of omega-3 free fatty acids (FFA) in human health have increased significantly in recent years. FFA properties have been studied extensively, including their potential therapeutic use in inflammatory conditions. However, omega-3 FFA's use as a supplementary treatment for OM has not been clinically tested. Preliminary evidence suggests that utilising FFA to manage OM could be a useful strategy for lesion management, assisting with healthy oral mucosa recovery. This review will describe the incidence, risk factors, biology of OM and the current treatment strategies, leading to a discussion of the utility of omega-3 FFA as a novel therapeutic agent for OM.

Claudin-7 down-regulation is an important feature in oral squamous cell carcinoma.

AIMS: Claudins, a large family of essential tight junction (TJ) proteins, are abnormally regulated in human carcinomas, especially claudin-7. The aim of this study was to investigate claudin-7 expression and alterations in oral squamous cell carcinoma (OSCC). METHODS AND RESULTS: Expression of claudin-7 was analysed in 132 cases of OSCC organized in a tissue microarray. Claudin-7 mRNA transcript was evaluated using real-time polymerase chain reaction and the methylation status of the promoter was also assessed. Claudin-7 was negative in 58.3% of the cases. Loss of claudin-7 protein expression was associated with recurrence (P = 0.019), tumour size (P = 0.014), clinical stage of OSCC (P = 0.055) and disease-free survival (P = 0.015). Down-regulation of the claudin-7 mRNA transcripts was observed in 78% of the cases, in accordance with immunoexpression. Analysis of the methylation status of the promoter region of claudin-7 revealed that treatment of O28 cells (that did not express claudin-7 mRNA transcripts) with 5-Aza-2'-Deoxycytidine (5-Aza-dC) led to the re-expression of claudin-7 mRNA transcript. CONCLUSION: Loss of claudin-7 expression is associated with important subcellular processes in OSCC with impact on clinical parameters.

Three photobiomodulation protocols in the prevention/treatment of radiotherapy-induced oral mucositis.

PURPOSE: To compare three Photobiomodulation protocols to prevent/treat oral mucositis associated to radiotherapy. METHODS: Seventy-three patients with cancer in oral cavity, oropharynx, and nasopharynx, who underwent RT with dose in facial fields equal or higher than 6000 cGy were randomized into three groups (mean RT dose = 66 cGy ±4.9). Protocols of Group 1 was 660 nm, 15 mW, 3.8 J/cm2, Group 2 660 nm, 25 mW, 6.3 J/cm2 both starting on the first day of radiotherapy, and group 3 660 nm, 15 mW, 3.8 J/cm2 for therapeutic purpose. The patients of group 1 and 2 were irradiated at 40 points daily covering non-keratinizing oral mucosa. The spot size (probe's tip surface size) was 0.040 cm2 for all groups. Oral mucositis was evaluated according to both WHO and NCI scales, and pain related to oral mucositis was scored using the VAS. RESULTS: Patients from group 1 presented with grade II oral mucositis later than groups 2 and 3 (p < 0.001). Moreover, groups 2 and 3 also presented with a mean higher of oral mucositis grade than group 1, p < 0.001. Pain scores were lower in group 1 (p = 0.002). CONCLUSIONS: The Photobiomodulation used in Group 1 was more effective than the protocols used in groups 2 and 3 in controlling the grade II oral mucositis intensity, and mean pain scores.

Efeitos da fotobiomodulação na imunidade de pacientes com mucosite oral / Effects of photobiomodulation in the immunity of patients with oral mucositis

A mucosite oral é a toxicidade aguda mais comum que acomete os pacientes submetidos à radioterapia em região de cabeça e pescoço. Das diversas formas de controle e tratamento desta lesão, a terapia com laser de baixa intensidade tem demonstrado, clinicamente, ser eficaz no controle da doença. Porém, não há um completo entendimento de como esta modalidade terapêutica atua na estimulação do sistema imunológico destes pacientes. Assim, o objetivo deste estudo foi investigar os mecanismos moleculares envolvidos na ação da laserterapia como tratamento da lesão no que se refere aos aspectos imunológicos. Para isso, foi realizado RT-PCR em tempo real (TaqMan® Gene Expression Assays), utilizando um painel de genes relacionados com o sistema imunológico, onde o nível de expressão gênica foi avaliado em 60 amostras de saliva coletadas de 23 pacientes portadores de câncer de boca, orofaringe e nasofaringe tratados por radioterapia exclusiva ou radioterapia associada à quimioterapia. Estes pacientes foram divididos em 3 grupos com diferentes intensidades de laser e modalidades terapêuticas: Grupo A preventivo (Potência de 15 mW e densidade de energia de 3,8 J/ cm²) com 8 pacientes e 23 amostras, Grupo B preventivo (Potência de 25 mW e densidade de energia de 6,3 J/ cm²) com 8 pacientes e 22 amostras e o Grupo C-controle- curativo (Potência de 15 mW e densidade de energia de 3,8 J/ cm²) com 7 pacientes e 15 amostras. Os resultados obtidos, através da análise de 52 genes, demonstraram que 42 genes apresentaram alteração de expressão ao longo da evolução da mucosite oral, sendo possível identificar que os genes CCL3, CCR7, SYK foram relacionados com a instalação e progressão da doença. Além disso, através das diferentes análises de expressão entre diferentes grupos estudados foi possível inferir que a modulação da expressão dos genes CCL2, CCL3, IL-1A, MIF, NFKB2, TNF podem dar respaldo molecular aos melhores resultados clínicos previamente observados na laserterapia preventiva, com potência de 15 mW e densidade de energia de 3,8 J/ cm². Assim, este trabalho gerou dados moleculares iniciais sobre os mecanismos da imunoregulação que estão envolvidos com a instalação e progressão da mucosite e com a fotobiomodulação usada no tratamento das lesões.

Oral mucositis is the most common acute toxicity affecting patients undergoing radiotherapy in head and neck. Of the different ways of controlling and treating this lesion, low intensity laser therapy has shown clinically effectiveness in controlling the disease. However, it is not fully understood how this therapeutic modality acts in stimulating the immune system of these patients. Therefore, the objective of this study was to investigate the molecular mechanisms involved in the action of laser therapy for the treatment of this lesion regarding the immunological aspects. For this purpose, real time RT-PCR (TaqMan Gene Expression Assays) was performed, using a panel of genes related to immune system, in which the level of gene expression was evaluated in 60 saliva samples collected from 23 patients harboring mouth, oropharynx and nasopharynx cancer treated by radiotherapy alone or radiotherapy combined with chemotherapy. These patients were divided into 3 groups with different laser intensities and treatment modalities: Group A preventive (power of 15 mW and energy density of 3.8 J / cm²) with 8 patients and 23 samples, preventive Group B (25 mW Power and energy density of 6.3 J / cm²) with 8 patients and 22 samples and Group C curative (15 mW of power and energy density of 3.8 J / cm²) with 7 patients and 15 samples. The results obtained, through the analysis of the 52 genes, showed that the expression of 42 genes was changed throughout the evolution of oral mucositis. It was possible to identify that the genes CCL3, CCR7, SYK were related to the onset and progression of the disease. Moreover, through different expression analyzes among the different groups studied it was possible to infer that gene expression modulation of CCL2, CCL3, IL-1A, MIF, NFKB2, TNF can give molecular backing to the best clinical results previously observed in preventive laser therapy, with power of 15 mW and energy density of 3.8 J / cm². Thus, this study has generated initial molecular data on the mechanisms of immunoregulation involved in the onset and progression of mucositis and with photobiomodulation being used as treatment for these lesions. the photobiomodulation used in treatment of injuries.