Connexin 43, Bcl-2, Bax, Ki67, and E-cadherin patterns in oral squamous cell carcinoma and its relationship with GJA1 rs12197797 C/G

Background: To our knowledge, there is no useful and accurate prognostic biomarker or biomarkers for patientswith oral squamous cell carcinoma (OSCC), a tumor with uncertain biological behavior, and unpredictable clinical progress. The purposes of this study were: a) to determine the expresión profile of Connexin 43, Bcl-2, Bax,E-cadherin, and Ki67 in patients with OSCC; b) identify the GJCA1 rs12197797 genotypic composition.Material and Methods: A cross-sectional study using genomic DNA and biopsy samples extracted from the oralmucosa with/without OSCC, older than 18 years, both genders, attended at Facultad de Odontología, UniversidadNacional Córdoba. Immunostaining for Cx43, Bcl-2, Bax, E-cadherin, and Ki67 and genotyping GJA1 rs12197797by RFLP were performed. Odds Ratio (95% CI), Spearman Coefficient were estimated. Mann-Whitney test wasapplied to analyze immunostaining between controls/cases (p <0.05 was set for statistical significance).Results: GG (mutant) was the most frequent genotype in patients with OSCC diagnosis (53.2%) in relation toCC “healthy” genotype (p=0.00487; OR=7.33; CI95% [1.1-54.7]). And, the allele G (mutant) had a presence in75.5% of OSCC patients. However, no significant association was observed between alleles C/G and diagnosis(p=0.0565). The heterozygous genotype was the most frequent in the patients of both groups Cx43 and E-cadherinmarkers were lower in OSCCs in relation to controls. Ki67 and Bcl-2 immunolabeling were high on OSCC, andBax immunomarker was diminished in OSCC.Conclusions: We hypothesized that the oral epithelium losses Connexin 43 and E-cadherin in the membrane, whichmodifies cell differentiation. The Ki67 and Bcl2 overexpression would increase the cell density in the tissue, by promoting proliferation and decreasing apoptosis. And, this study shows evidence that patients who carry on allele G ofGJA1rs12197797 could be at risk of developing OSCC. (AU)

Exfoliative cytology as a tool for monitoring pre-malignant and malignant lesions based on combined stains and morphometry techniques.

BACKGROUND: Prevention and early diagnosis have the greatest potential for public health and are the most effective method in the long-term to control oral cancer. The aim was to apply PAP staining together with AgNOR staining and morphometric analysis in oral exfoliative cytology, to determine the sensitivity and specificity of these methods in the detection of malignant changes for the purposes of both initial population monitoring and follow-up. METHODS: AgNOR, Papanicolau, and morphometric tests were conducted in samples of patients with oral cancer, oral potentially malignant disorders and controls (opposite side of lesions). Specificity and sensitivity values for each stain method and the curve under ROC area were estimated. RESULTS: The diagnostic variables which allowed greatest accuracy in identifying malignancy relative to the healthy control were cluster (76.92%), satellite (75.64%), and total (90%). The diagnosis was seen to be associated with PAP and total AgNOR, total AgNOR and PAP, total AgNOR and satellites and clusters, and total AgNOR nuclear area/cytoplasmic area ratio. CONCLUSIONS: The total number of AgNOR is a reliable marker for detecting neoplastic cells; this method increases sensitivity and specificity by decreasing the likelihood of false negatives or positives, as the accuracy obtained was 90%. It is also a low-cost, non-invasive, simple methodology that can be recommended to help the early detection of oral cancer and monitoring of patients with a first diagnosis of cancer.

Estudio citomorfométrico de pacientes con cáncer y desórdenes potencialmente malignos de la cavidad bucal / Citomorphometric study of patients with cancer and potentially malignant disorders of the oral cavity

La detección temprana de lesiones bucales promete aumentar sobrevivencia y reducir la morbilidad de los pacientes que sufren esta condición. Un método alternativo para el examen de lesiones en la cavidad bucal, es la citología. Objetivo: valorar las relaciones del tamaño núcleo/citoplasma de células de mucosa bucal sana, de lesiones bucales potencialmente malignas y de cáncer bucal, obtenidas con citología exfoliativa utilizando citomorfometría. Material y método: Se realizó citología a 22 pacientes, de ambos sexos, con edades entre 23 y 81 años utilizando cytobrush. Las muestras se dividieron en tres grupos: 1) citologías de pacientes con lesiones de cáncer bucal (n=7); 2) citologías de pacientes con desórdenes bucales potencialmente malignos (leucoplasia y líquen) (n=15); 3) citologías del lado sano de pacientes con lesiones de cáncer y desórdenes bucales potencialmente malignos (grupo control) (n=15). Se seleccionaron 30 células de cada paciente y se midió el área nuclear (AN), la citoplasmática (AC) y se calculó la relación AN/AN. Se utilizó el Test de Kruskal Wallis y el Sofware estadístico Infostat. Resultados: No se encontraron diferencias significativas entre los tres grupos estudiados al valorar la relación AN/AC. Teniendo en cuenta el género, se observó que en las mujeres se diferenciaron significativamente las células del grupo control de las de los grupos de lesiones. En el género masculino se observaron diferencias significativas entre los tres grupos celulares. No hubo diferencias significativas entre los diferentes grupos etarios .Discusión: A pesar de las diferencias significativas entre los géneros, no es posible hacer una buena separación de los tres grupos de estudio, utilizando solamente citología exfoliativa.

Early detection of bucal lesions promises to increase survival and to reduce morbidity in patients suffering from this condition. An alternative method for examining lesions in the bucal cavity is cytology. Objective: To assess by cytomorphometry the relationship of size nucleus / cytoplasm of superficial cells from healthy bucal mucosa, from potentially malignant bucal disordes lesions and bucal cancer using exfoliative cytology. Material and methods: PAP smear was performed in 22 patients of both gender, aged between 23 and 81. Cytobrush was used. The samples were divided into three study groups: 1) smears of patients with bucal cancer lesions (n = 7), 2) smears of patients with potentially malignant bucal disordes lesions (n = 15), lesions considered were leukoplakia and lichens ; 3) (control group) (n=15): smears of the healthy side of patients with cancer and potentially malignant bucal disordes lesions.Thirty cells were selected for each patient and the nuclear area (NA), cytoplasmic area (CA) were measured and the ratio NA / CA was calculated. We used Kruskal Wallis and Statistical Software InfoStat. Results: No significant differences were found between the three groups studied the relationship NA / CA was assesed . When comparing cells from each of the three groups considering the gender of the patients separately, we found that in women differ significantly from the control group cells and groups of injuries, we could not discriminate between cells obtained from potentially malignant bucal disordes lesions and cancer lesions . In the male gender differences were significant among the three cell groups. Discusion: Despite the significant gender differences, we could not difference the three study groups using only exfoliative cytology.