The Role of SOX2 and SOX9 in Radioresistance and Tumor Recurrence.

Head and neck squamous cell carcinoma (HNSCC) exhibits considerable variability in patient outcome. It has been reported that SOX2 plays a role in proliferation, tumor growth, drug resistance, and metastasis in a variety of cancer types. Additionally, SOX9 has been implicated in immune tolerance and treatment failures. SOX2 and SOX9 induce treatment failure by a molecular mechanism that has not yet been elucidated. This study explores the inverse association of SOX2/SOX9 and their distinct expression in tumors, influencing the tumor microenvironment and radiotherapy responses. Through public RNA sequencing data, human biopsy samples, and knockdown cellular models, we explored the effects of inverted SOX2 and SOX9 expression. We found that patients expressing SOX2LowSOX9High showed decreased survival compared to SOX2HighSOX9Low. A survival analysis of patients stratified by radiotherapy and human papillomavirus brings additional clinical relevance. We identified a gene set signature comprising newly discovered candidate genes resulting from inverted SOX2/SOX9 expression. Moreover, the TGF-ß pathway emerges as a significant predicted contributor to the overexpression of these candidate genes. In vitro findings reveal that silencing SOX2 enhances tumor radioresistance, while SOX9 silencing enhances radiosensitivity. These discoveries lay the groundwork for further studies on the therapeutic potential of transcription factors in optimizing HNSCC treatment.

MiRNAs that target amyloid precursor protein processing machinery in extracellular vesicles and particles derived from oral squamous cells carcinoma.

BACKGROUND: Considering that microRNAs (miRNAs), extracellular vesicles and particles (EVPs) and the amyloid precursor protein (APP) processing have been shown to be altered in oral squamous cells carcinoma (OSCC), it is possible that miRNAs that target APP processing pathways in EVPs are impacted in tumor cells. Our aim was to evaluate miRNAs that target APP itself or disintegrin and metalloproteinase domain 10 (ADAM10), which generate a trophic compound, sAPPα, in EVPs derived from OSCC cell lines, an aggressive and non-invasive, compared to normal keratinocytes. METHODS: We used two OSCC cell lines, an aggressive human oral squamous cell carcinoma cell line (SCC09) and a less aggressive cell line (CAL27) compared with a keratinocyte lineage (HaCaT). Cells were maintained in cell media, from which we isolated EVPs. EVPs were evaluated regarding their size and concentration using Nanotracking Analysis. We measured the levels of miRNAs which had as potential downstream target APP or ADAM10, specifically miR-20a-5p, miR-103a-3p, miR-424-5p, miR-92b-3p, miR-31-5p, and miR-93-5. RESULTS: There were no differences on size distributions and concentration of isolated EVPs. OSCC cell lines-derived EVPs miR-20a-5p, miR-92b-3p, and miR-93-5p were upregulated in comparison to HaCaT-derived EVPs; while miR-31-5p was reduced in EVPs obtained from CAL27 cells. CONCLUSION: Our results indicate changes in miRNAs that target APP machinery processing in EVPs derived from OSCC cell lines of different aggressiveness, which may be involved with abnormal miRNA expression in OSCC tissue and/or releasing tumor suppressor miRNA.

Asymmetric mitosis contributes to different migratory performance in sister cells.

In cancer, cell migration contributes to the spread of tumor cells resulting in metastasis. Heterogeneity in the migration capacity can produce individual cells with heightened capacity leading to invasion and metastasis. Our hypothesis is that cell migration characteristics can divide asymmetrically in mitosis, allowing a subset of cells to have a larger contribution to invasion and metastasis. Therefore, our aim is to elucidate whether sister cells have different migratory capacity and analyze if this difference is defined by mitosis. Through time-lapse videos, we analyzed migration speed, directionality, maximum displacement of each trajectory, and velocity as well as cell area and polarity and then compared the values between mother-daughter cells and between sister cells of three tumor cell lines (A172, MCF7, SCC25) and two normal cell lines (MRC5 and CHO·K1 cells). We observed that daughter cells had a different migratory phenotype compared to their mothers, and one single mitosis is enough for the sisters behave like nonrelated cells. However, mitosis did not influence cell area and polarity dynamics. These findings indicates that migration performance is not heritable, and that asymmetric cell division might have an important impact on cancer invasion and metastasis, by producing cells with different migratory capacity.

Cell contractility drives mechanical memory of oral squamous cell carcinoma.

Matrix stiffening is ubiquitous in solid tumors and can direct epithelial-mesenchymal transition (EMT) and cancer cell migration. Stiffened niche can even cause poorly invasive oral squamous cell carcinoma (OSCC) cell lines to acquire a less adherent, more migratory phenotype, but mechanisms and durability of this acquired "mechanical memory" are unclear. Here, we observed that contractility and its downstream signals could underlie memory acquisition; invasive SSC25 cells overexpress myosin II (vs. noninvasive Cal27 cells) consistent with OSCC. However, prolonged exposure of Cal27 cells to a stiff niche or contractile agonists up-regulated myosin and EMT markers and enabled them to migrate as fast as SCC25 cells, which persisted even when the niche softened and indicated "memory" of their prior niche. Stiffness-mediated mesenchymal phenotype acquisition required AKT signaling and was also observed in patient samples, whereas phenotype recall on soft substrates required focal adhesion kinase (FAK) activity. Phenotype durability was further observed in transcriptomic differences between preconditioned Cal27 cells cultured without or with FAK or AKT antagonists, and such transcriptional differences corresponded to discrepant patient outcomes. These data suggest that mechanical memory, mediated by contractility via distinct kinase signaling, may be necessary for OSCC to disseminate.

Applicability of autofluorescence and fluorescent probes in the trans-surgical of oral carcinomas: A systematic review.

Oral cancer represents an important health problem, as it is the sixth most common type of cancer in the world and is associated with high rates of morbidity and mortality. The treatment considered the gold standard for this type of tumor is surgical resection with negative margins, with a distance of at least 5 mm from the tumor. This procedure is strongly associated with local control and disease-specific survival, however, in many cases, large amounts of healthy tissue are removed, resulting in surgical defects, compromising various functions and directly affecting the individual's quality of life. From this perspective, this systematic review aimed to evaluate the use of autofluorescence and fluorescent probes as potential adjuvant techniques to facilitate the delineation of surgical margins for oral cancers. A comprehensive search was performed in Pubmed, Scopus, Web of Science, LIVIVO, Embase, ProQuest Open Access Dissertations & Theses, Open Access Theses and Dissertations, and DART Europe databases, where 1948 articles were found. After the different stages of critical evaluation, 15 articles were selected, eligible for the inclusion criteria. Of these, 7 articles used autofluorescence, 7 used fluorescent probes and 1 article used both methods. As for autofluorescence, the most used device was the VELScope, and indocyanine green was the most used probe. Compared to histopathology, autofluorescence did not obtain significant and/or superiors results. In contrast to fluorescent probes that, most articles showed a good performance of margins during surgical resection, making them a promising alternative. However, it is still necessary to carry out the analysis of more articles, with more significant samples and sensitivity and specificity data to qualify the results.

Applicability of autofluorescence and fluorescent probes in early detection of oral potentially malignant disorders: A systematic review and meta-data analysis.

Oral potentially malignant disorders (OPMD) represent a group of lesions with increased risk for malignant transformation. The management of such injuries is based on surgical treatment or detailed follow-up throughout the patient's lifetime. This systematic review and meta-analysis investigated and critically evaluated the use of autofluorescence and fluorescent probes as potential techniques for the early detection of OPMD. A comprehensive search was performed on Pubmed, Scopus, Web of Science and LIVIVO databases. The gray literature was also consulted and included Google Scholar, Proquest and Open gray databases. 2715 articles were retrieved, and after the different stages of critical evaluation, were reduced to 25 articles that fully met the inclusion criteria. VELscope® was the most used equipment for autofluorescence, while aminolevulinic acid (5-ALA) was the main representative of the probes. The meta-analysis performed included 10 articles that used VELscope® as a method to detect oral disorders. A 95% confidence interval (CI) with a p value significance <0.05 was considered as a criterion for the statistical analysis. The combined sensitivity was 74% (CI95 60-76%, p = 0.0001) and the specificity was 57% (CI95 52-60%, p = 0.0000). The inclusion of these adjunct methods in clinical practice is very promising, since they are able to help both the clinician and the specialist in the early detection of potentially malignant oral disorders, favoring a better prognosis. However, it is still necessary to carry out further studies, with the aim of establishing a protocol for use and qualification of results.

The role of tumor acidification in aggressiveness, cell dissemination and treatment resistance of oral squamous cell carcinoma.

AIMS: To investigate the role of tumor acidification in cell behavior, migration, and treatment resistance of oral squamous cell carcinoma (OSCC). MAIN METHODS: The SCC4 and SCC25 cell lines were exposed to acidified (pH 6.8) cell culture medium for 7 days. Alternatively, a long-term acidosis was induced for 21 days. In addition, to mimic dynamic pH fluctuation of the tumor microenvironment, cells were reconditioned to neutral pH after experimental acidosis. This study assessed cell proliferation and viability by sulforhodamine B and flow cytometry. Individual and collective cell migration was analyzed by wound healing, time lapse, and transwell assays. Modifications of cell phenotype, EMT induction and stemness potential were investigated by qRT-PCR, western blot, and immunofluorescence. Finally, resistance to chemo- and radiotherapy of OSCC when exposed to acidified environmental conditions (pH 6.8) was determined. KEY FINDINGS: The exposure to an acidic microenvironment caused an initial reduction of OSCC cells viability, followed by an adaptation process. Acidic adapted cells acquired a mesenchymal-like phenotype along with increased migration and motility indexes. Moreover, tumoral extracellular acidity was capable to induce cellular stemness and to increase chemo- and radioresistance of oral cancer cells. SIGNIFICANCE: In summary, the results showed that the acidic microenvironment leads to a more aggressive and treatment resistant OSCC cell population.

The anti-tumor effects of imidazolium salts on oral squamous cell carcinoma.

BACKGROUND: Imidazolium salts (IS), ionic derivatives of neutral imidazoles, have properties that can be adjusted by structural modifications to their cations and anions, which makes this particular class of compounds a promising option for developing biologically active compounds. The anti-tumor effects of the IS 1-n-butyl-3-methylimidazolium chloride (C4 MImCl), 1-n-decyl-3-methylimidazolium chloride (C10 MImCl), 1-n-hexadecyl-3-methylimidazolium chloride (C16 MImCl), 1-n-hexadecyl-2,3-dimethylimidazolium chloride (C16 M2 ImCl), 1-n-octadecyl-3-methylimidazolium chloride (C18 MImCl), 1-n-hexadecyl-3-methylimidazolium methanesulfonate (C16 MImMeS), and 1-n-hexadecyl-2,3- dimethylimidazolium methanesulfonate (C16 M2 ImMeS) on oral squamous cell carcinoma (OSCC) have been studied here. METHODS: Oral squamous cell carcinoma cells (CAL27) were incubated with increasing IS doses and then submitted to proliferation (2D), cell death (2D) and spheroid assay (3D). RESULTS: The IS anti-tumor effect was dependent on both its N-alkyl chain length and anion, whereby C16 MImCl proved to be more effective in combination for inhibiting cell proliferation and cell-cell adhesion, outperforming the methylated C16 M2 ImCl derivative and, most importantly, the gold standard-cisplatin. In addition, C16 MImCl had little effect on keratinocytes and more pronounced effects on more aggressive tumor cells. It also exhibited similar effects on inducing cell death when compared to Cisplatin. This compound spread to a greater area of the tumor sphere and produced an enhanced number of apoptotic and necrotic cells in the tumor cell line, demonstrating only a small rise in the healthy cells. CONCLUSION: These data indicate that the effect of C16 MlmCl on OSCC is promising, as it is selective for cancer cells.

Use of autofluorescence and fluorescent probes as a potential diagnostic tool for oral cancer: A systematic review.

INTRODUCTION: The prognosis of patients with Oral squamous cell carcinoma (OSCC) are directly related to the stage of development of the tumor at the time of diagnosis, but it is estimated an average delay in diagnosis of 2-5 months. New non-invasive techniques for the early diagnosis of OSCC are being developed, such as methodologies to detect spectral changes of tumor cells. We conducted a systematic review to analyze the potential use of autofluorescence and/or fluorescent probes for OSCC diagnosis. MATERIAL AND METHODS: Four databases (PubMed, Scopus, Embase and Web of Science) were used as research sources. Protocol was registered with PROSPERO. It was included studies that evaluated tissue autofluorescence and/or used fluorescent probes as a method of diagnosing and/or treatment of oral cancer in humans. RESULTS: Forty-five studies were selected for this systematic review, of which 28 dealt only with autofluorescence, 18 on fluorescent probes and 1 evaluated both methods. The VELscope® was the most used device for autofluorescence, exhibiting sensitivity (33%-100%) and specificity (12%-88.6%). 5-Aminolevulinic acid (5-ALA) was the most used fluorescent probe, exhibiting high sensitivity (90%-100%) and specificity (51.3%-96%). Hypericin, rhodamine 6 G, rhodamine 610, porphyrin and γ-glutamyl hydroxymethyl rhodamine green have also been reported. CONCLUSION: Thus, the autofluorescence and fluorescent probes can provide an accurate diagnosis of oral cancer, assisting the dentist during daily clinical activity, but it is not yet possible to suggest that this method may replace histopathological examination.

Flavonoids as anticancer therapies: A systematic review of clinical trials.

Flavonoids have been proposed as potential chemotherapeutic agents because they are toxic against cancer cells but not harmful to healthy cells. This systematic review analyzed flavonoid effectiveness in human cancer chemotherapy. Overall, 22 phase II and 1 phase III clinical trials (PubMed, Scopus, and Web of Science) that used flavonoids as a single agent or combined with other therapeutics against hematopoietic/lymphoid or solid cancer published by January 2019 were selected for analysis. Flavopiridol was the most commonly used flavonoid (at a dose of 50-mg/m2 IV) for all tumor types. Aside from the relatively low rate of complete response (CR) or partial response (PR) with any administration protocol, flavonoids showed higher positive outcomes for hematopoietic and lymphoid tissues (140 patients with CR and 88 with PR among 615 patients in 11 trials) than for solid tumors (4 patients with CR and 21 with PR among 525 patients in 12 trials). However, because of the high variety in administration schedule, more studies are needed to further understand how flavonoids can promote positive outcomes for cancer patients.

Periodontal Disease Impairs Muscle Recovery by Modulating the Recruitment of Leukocytes.

The purpose of this study is to analyze the impact of periodontal disease (PD) associated with physical exercise on inflammatory mediators and muscle repair. Twenty-four Wistar rats were divided into four groups: control (SH), healthy trained (TH), sedentary with PD (SP), and trained with PD (TP). PD was induced in groups SP and TP while the trained groups performed treadmill exercises for 8 weeks. For the analysis of IL-6, IL-10, TNF-α, and leukocyte count, we collected blood samples. Cryolesions were induced in the tibialis anterior and gastrocnemius, which were analyzed for morphological changes. The presence of PD modified leukocyte counts, while exercise showed an additive role. PD increased levels of IL-6, IL-10, and TNF-α, and physical exercise changed only values of IL-10. The association between physical exercise and PD was responsible for an increased concentration of leukocytes in the region of the inflammation. Serum levels of inflammatory markers were modified by PD and, when combined with exercise, may negatively modulate inflammation. The association between PD and physical exercise showed the most significant changes in the number of inflammatory cells and may negatively influence the process of muscle repair.

Keratinocyte Cytotoxicity of Peracetic Acid Used as Sterilizing Agent for Implant Scaffolds.

Peracetic acid (PAA) has been used to sterilize biomaterial scaffolds and allografts before their implantation. Although the antimicrobial effectiveness of PAA is widely known, there are no studies investigating its cytotoxicity on keratinocytes. This study aimed to investigate the cytotoxicity of PAA concentrations on keratinocytes by growing HaCaT cells in culture medium. Different concentrations of PAA (control-untreated, 0.01, 0.1, 1, 10, 100, 200, 400, 800, 1200, 1600, and 2000 ppm) were added to the culture wells and allowed to be in direct contact with cells for up to 24 hours. Cytotoxicity was quantitatively and qualitatively determined by cell viability assay and analysis of morphological changes. Statistical analysis was performed with 1-way analysis of variance and Tukey test at 5% significance. Cells treated with 0.01 and 0.1 ppm followed the same morphological pattern of untreated cells, whereas cells treated with 1.0 ppm presented about 20% of floating cells and dark cytoplasmic granules. More than 50% of the cells treated with 10 and 100 ppm were destroyed, whereas the attached ones showed unclear and interrupted cell membranes. Concentrations of 1 ppm or greater had less than 64.4% of viable cells compared with the control group. This study concluded that exposure of keratinocytes to concentrations of 1 ppm or greater of PAA resulted in strong cytotoxic effects.

Effects of low molecular weight heparin on alveolar bone loss in wistar rats

Abstract This study aimed to assess the effects of low molecular weight heparin (LMWH) on alveolar bone loss (ABL), blood count, and counting of megakaryocytes and adipocytes in male Wistar rats. Forty male 60-day Wistar rats were randomly divided into four groups: Control (C), Periodontal Disease (PD), Heparin (Hp) and Heparin + Periodontal Disease (Hp+PD). LMWH was applied for 60 days at doses of 1 ml/kg/day. Blood samples were collected at baseline, 30 and 60. On day-49, PD and Hp+PD groups were subjected to ligature-induced periodontitis around second upper right molar. The left side was assessed as spontaneous alveolar bone loss. Mean ABL in the side with ligature showed significantly different between C (0.35±0.07 mm) and Hp+DP (0.49±0.09 mm) groups (p<0.001), between PD (0.55±0.11 mm) and Hp (0.32±0.06 mm) groups (p<0.001) and between Hp and Hp+DP groups (p<0.001). No significant differences were found among groups for ABL in the side without ligature. Animal weight, food intake, and water consumption showed no statistically significant difference among groups. Megakaryocytes and adipocytes were counted using optical microscopy and no statistically significant differences were found. Within-groups, there were an increase and a decrease, respectively, in the counting of lymphocytes (p=0.005 for C and p=0.009 for Hp+PD groups only) and leukocytes (p=0.003 for C, p=0.001 for PD, p=0.002 for Hp, and p<0.001 for Hp+PD groups). There was no decrease in the number of platelets in the three collection periods. LMWH was not able to affect ABL, but it may change the blood counting, especially increasing lymphocytes.

Resumo O presente estudo objetivou verificar o efeito da heparina de baixo peso molecular (HBPM) sob a perda óssea alveolar (POA), contagem de células sanguíneas, megacariócitos e adipócitos em ratos Wistar machos. Quarenta ratos Wistar de 60 dias foram randomicamente divididos em quatro grupo: Controle (C), Doença Periodontal (DP), Heparina (Hp) e Heparina + Doença Periodontal (Hp+DP). HBPM foi aplicada durante 60 dias em doses de 1 mL/kg/dia. Coletas sanguíneas foram realizadas nos dias 0, 30 e 60. No dia 49, os grupos DP e Hp+DP receberam indução de doença periodontal por ligadura ao redor do segundo molar superior direito. No lado esquerdo, verificou-se perda óssea alveolar espontânea. A média de POA no lado com ligadura mostrou-se estatisticamente diferente entre os grupos C (0,35±0,07 mm) e Hp+PD (0,49±0,09 mm) (p<0,001), entre os grupos DP (0,55±0,11 mm) e Hp (0,32±0,06 mm) (p<0,001) e entre os grupos Hp e Hp+DP (p<0,001). Nenhuma diferente significativa foi observada entre os grupos no lado sem ligadura. Peso dos animais, consumo de ração e ingestão de água não mostraram diferenças significativas entre os grupos. Megacariócitos e adipócitos foram contados por microscopia óptica e nenhuma diferença significativa foi encontrada. Dentro dos grupos, houve um aumento e uma diminuição, respectivamente, na contagem de linfócitos (p=0,005 no grupo C e p=0,009 no grupo Hp+DP somente) e leucócitos (p=0,003 no grupo C, p=0.001 no grupo DP e p=0,002 no grupo Hp e Hp+DP). Não houve diminuição no número de plaquetas nos três períodos de coleta. HBPM não foi capaz de modificar a POA, porém modificou a contagem de células sanguíneas, especialmente aumentando o número de linfócitos.

The amyloid precursor protein (APP) processing as a biological link between Alzheimer's disease and cancer.

Aging is a risk factor for several illnesses, such as Alzheimer's Disease and various cancers. However, an inverse correlation between malignancies and Alzheimer's Disease has been suggested. This review addressed the potential role of non-amyloidogenic and amyloidogenic pathways of amyloid precursor protein processing as a relevant biochemical mechanism to clarify this association. Amyloidogenic and non-amyloidogenic pathways have been related to Alzheimer's Disease and certain malignancies, respectively. Several known molecules involved in APP processing, including its regulation and final products, were summarized. Among them some candidate mechanisms emerged, such as extracellular-regulated kinase (Erk) and protein kinase C (PKC). Therefore, the imbalance of APP processing may be involved with the negative correlation between cancer and Alzheimer Disease.

NADPH-oxidase-derived ROS alters cell migration by modulating adhesions dynamics.

BACKGROUND INFORMATION: Cell migration requires the coordinated activation of structural and signalling molecules, such as the RhoGTPase Rac1. It is known that the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex assembly, which generates reactive oxygen species (ROS) at the cell membrane, also relies on Rac1 activation, indicating a possible effect of ROS during cell migration. In this study, we evaluated the effect of NADPH-oxidase-derived ROS on the migration process. RESULTS: Using time-lapse videos of CHO.K1 cells plated on fibronectin (2 µg/ml) or collagen (5 µg/cm2 ), we observed that depletion of ROS by N-acetyl-cysteine (NAC, 10 mM), an unspecific antioxidant, or diphenyliodonium (DPI, 10 µM), a NADPH-oxidase inhibitor, induced a ∼50% decrease in migration speed and severely impacted migration directionality. Then, we analysed the effects of NADPH oxidase on three migratory events: protrusion rate, adhesion process and signalling pathways related to cell migration. DPI induced an increase of ∼3 protrusion/cell, which were 2× faster but had a ∼50% retraction when compared with control. By pull-down assay, we observed no changes on Rac1 activation, indicating that ROS-mediated effects were related to downstream molecules, such as adhesion-related molecules. A reduction of the adhesion marker FAK-Y397 levels in cells treated with NAC and DPI was observed. In order to analyse adhesion dynamics, CHO.K1 cells transfected with paxillin-GFP analysed with total internal reflectance fluorescence (TIRF) indicated that DPI (5 µM) induced larger adhesions when compared with control. CONCLUSION: These results indicate that the local generation of NADPH-oxidase-derived ROS can modulate cell migration due to changes on adhesion dynamics and signalling. SIGNIFICANCE: This study highlights the physiological requirement of ROS for cell migration and the potential use of these molecules as targets to modulate the cell migration process at different diseases.

Effects of diabetes and hypertension on oral mucosa and tgfß1 salivary levels

Abstract The aim of this study was to investigate salivary levels of TGFβ1 and proliferation/ maturation of epithelial mucosa cells in diabetic and hypertensive patients. Design: in this cross-sectional study, whole stimulated saliva and oral mucosa exfoliative cytology specimens were collected from 39 patients that were healthy (control, n=10) or presented history of arterial hypertension (HAS, n=9), diabetes mellitus (DM, n=10) or both (DM+HAS, n=10). Salivary flow rate (SFR), TGFβ1 level in saliva, AgNORs and the epithelial maturation were evaluated. Non-parametric Kruskal-Wallis test, followed by Dunn's multiple comparison post-test and the Spearman test correlation analysis were used. SFR showed a significant decreased in DM and DM+HAS (0.47±0.11 and 0.64±0.43 mL/min) when compared to control (1.4±0.38 mL/min). DM+HAS presented the highest value of TGFβ1 concentration (24.72±5.89 pg/mL). It was observed a positive correlation between TGFβ1 and glycaemia (R=0.6371; p<0.001) and a negative correlation between TGFβ1 and saliva (R=-0.6162; p<0.001) and glycaemia and SFR (R=-0.5654; P=0.001). AgNORs number and status of maturation of mucosa cells were similar for all conditions. DM and DM+HAS presented the lowest SFR, which correlated with increased TGFβ1 levels. Despite the higher TGFβ1 secretion it was not observed changes in the morphology or proliferation of epithelial cells when diabetes or hypertension was present.

Resumo O objetivo deste estudo foi investigar os níveis de TGFβ1 na saliva e a proliferação/maturação das células epiteliais da mucosa em paciente diabéticos e hipertensos. Neste estudo transversal, saliva estimulada e amostras de citologia exfoliativa de mucosa oral foram coletadas de um total de 39 pacientes que se apresentavam saudáveis (controle, n=10) ou com história de hipertensão arterial (HAS, n=9), diabetes mellitus (DM, n=10) ou ambos (DM+HAS, n=10). Taxa de fluxo salivar (SFR), níveis de TGFβ1 na saliva, AgNORs e maturação epitelial foram avaliados. Teste não-paramétrico de Kruskal-Wallis, seguido de comparação múltipla de Dunn e correlação de Spearman foram utilizados para as análises. SFR diminuiu significantemente em DM e DM+HAS (0,47±0,11 e 0,64±0,43 mL/min) quando comparado ao controle (1,4±0,38 mL/min). DM+HAS apresentou os maiores valores de concentração de TGFβ1 (24,72±5,89 pg/mL). Foi observada uma correlação positiva entre TGFβ1 e glicemia (R=0,6371; p<0,001) e uma correlação negativa entre TGFβ1 e saliva (R=-0,6162; p<0,001) e glicemia e SFR (R=-0,5654; p=0,001). Número de AgNORs e o padrão da maturação das células epiteliais foram similares entre os todos grupos. DM e DM+HAS apresentaram os menores valores de SFR, os quais foram correlacionados com o aumento nos níveis de TGFβ1. Apesar da maior secreção de TGFβ1, não foram observadas mudanças na morfologia ou proliferação das células epiteliais quando o paciente apresentava diabetes ou hipertensão.

Focal adhesion kinases in head and neck squamous cell carcinoma.

BACKGROUND: Focal adhesion kinase (FAK) phosphorylation contributes to the regulation of growth factors that promote cellular adhesion, mobility, and survival, being a key factor in tumor development. The objective of this study was to evaluate the immunohistochemical expression patterns of FAK and its phosphorylated forms, FAK Tyr-576 and FAK Tyr-925, in head and neck squamous cell carcinoma and non-neoplastic adjacent epithelial tissue (AE). METHODS: The percentage of immunohistochemistry stained cells and its correlation with clinicopathological variables and prognosis were determined using samples from 54 patients. RESULTS: FAK, FAK Tyr-576, and FAK Tyr-925 overexpression was observed in tumor zones and AE. FAK Tyr-576 immunostaining showed a relationship with tumor clinicopathological parameters. Moreover, positive immunostaining of FAK Tyr-576 in AEsue was associated with patients prognoses. CONCLUSIONS: Increased expression of FAK Tyr-576 could enable identification of tumors with a more aggressive behavior and epithelial alterations before the appearance of clinical or histological manifestations.

Macrophages and prognosis of oral squamous cell carcinoma: A systematic review.

Oral Squamous Cell Carcinoma (OSCC) presents a tumor microenvironment rich in inflammatory cells. Depending on the stimulus, macrophages can polarize in M1 or M2 profile, where M1 acts as proinflammatory and antitumor, and M2 is anti-inflammatory and shows protumor activity. Several studies have shown that macrophages are important to the prognosis of patients with different types of cancer. Our aim was to conduct a systematic review to evaluate the role of macrophages in the prognosis of OSCC patients. A search in the Pubmed, Scopus, and ISI Web of Knowledge database was performed, and it was included only studies that evaluated the importance of macrophages in the prognosis of OSCC patients. From initial 286 articles, 14 fully attended the inclusion criteria. In the majority of the articles, it was evaluated only CD68, a panmacrophage marker, or CD163, a M2 marker. Only one article evaluated the M1 marker, CD11c. Besides, 5 articles analyzed the presence of macrophages in different areas of the tumor. Higher concentrations of CD68 and CD163 were associated with worse survival. In conclusion, macrophages are important to OSCC patients' prognosis; however, it is necessary to address in which tumor region the presence of polarized macrophage is more important to the outcome.

Low Doses of Curcuma longa Modulates Cell Migration and Cell-Cell Adhesion.

Cell invasion and metastasis are involved in clinical failures in cancer treatment, and both events require the acquisition of a migratory behavior by tumor cells. Curcumin is a promising natural product with anti-proliferative activity, but its effects on cell migration are still unclear. We evaluated the effects of curcumin on the proliferation, apoptosis, migration, and cell-cell adhesion of keratinocyte, oral squamous cell carcinoma (OSCC), and fibroblast cell lines, as well as in a xenograft model of OSCC. Curcumin (2 µM) decreased cell proliferation in cell lines with mesenchymal characteristics, while cell death was detected only at 50 µM. We observed that highly migratory cells showed a decrease on migration speed and directionality when treated with 2 or 5 µM of curcumin (50% and 40%, respectively, p < 0.05). Using spheroids, we observed that curcumin dose dependently decreased cell-cell adhesion, especially on tumor-derived spheroids. Also, in a xenograft model with patient-derived OSCC cells, the administration of curcumin decreased tumor growth and aggressiveness when compared with untreated tumors, indicating the potential antitumor effect in oral cancer. These results suggest that lower doses of curcumin can influence several steps involved in tumorigenesis, including migration properties, suggesting a possible use in cancer therapy. Copyright © 2017 John Wiley & Sons, Ltd.

Aging process alters hippocampal and cortical secretase activities of Wistar rats.

A growing body of evidence has demonstrated amyloid plaques in aged brain; however, little attention has been given to amyloid precursor protein (APP) processing machinery during the healthy aging process. The amyloidogenic and non-amyloidogenic pathways, represented respectively by ß- and α-secretases (BACE and TACE), are responsible for APP cleavage. Our working hypothesis is that the normal aging process could imbalance amyloidogenic and non-amyloidogenic pathways specifically BACE and TACE activities. Besides, although it has been showed that exercise can modulate secretase activities in Alzheimer Disease models the relationship between exercise effects and APP processing during healthy aging process is rarely studied. Our aim was to investigate the aging process and the exercise effects on cortical and hippocampal BACE and TACE activities and aversive memory performance. Young adult and aged Wistar rats were subjected to an exercise protocol (20min/day for 2 weeks) and to inhibitory avoidance task. Biochemical parameters were evaluated 1h and 18h after the last exercise session in order to verify transitory and delayed exercise effects. Aged rats exhibited impaired aversive memory and diminished cortical TACE activity. Moreover, an imbalance between TACE and BACE activities in favor of BACE activity was observed in aged brain. Moderate treadmill exercise was unable to alter secretase activities in any brain areas or time points evaluated. Our results suggest that aging-related aversive memory decline is partly linked to decreased cortical TACE activity. Additionally, an imbalance between secretase activities can be related to the higher vulnerability to neurodegenerative diseases induced by aging.