Evaluation of p16ink4a expression and presence of HPV-16 DNA by Real time PCR in patients with oral leukoplakia
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Data
2019-08-12
Autores
Biss, Stephanye Pinto [UNESP]
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Universidade Estadual Paulista (Unesp)
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Objetivo: Avaliar a expressão do p16Ink4a por imunohistoquímica e a presença do HPV16 pela Real time PCR em tecido fresco, plasma e saliva de pacientes com leucoplasia bucal (LB) e hiperplasia fibrosa inflamatória (HFI). Material e métodos: Foram incluídos 67 pacientes com o diagnóstico de LB e 44 pacientes com diagnóstico de HFI no estudo. Foram coletados dados sociodemográficos, clinicopatológicos, amostras de tecido fresco, sangue e saliva, que foram armazenados em um freezer a -80o C para realização de análise molecular. Também foi utilizado o tecido parafinizado para realização da imunohistoquímica para a p16Ink4a. As amostras de materiais biológicos obtidos dos pacientes foram submetidas à detecção do DNA do HPV-16 pela Real time PCR. O tecido parafinizado dos mesmos pacientes foram utilizados para avaliar a expressão da p16Ink4a pela imunohistoquímica. Resultados: Dos 67 pacientes incluídos de LB no estudo, 55,2% eram do sexo masculino com uma média de idade de 57,1 anos. Os pacientes idosos (>45 anos) compuseram 86,6% da amostra. As localizações anatômicas mais acometidas por LB foram a mucosa jugal (35,8%) e língua (20,9%). Sobre o tabagismo, 71,8% dos pacientes eram fumantes, onde a maioria (47,8%) foi classificada como tabagista leve. Em relação ao consumo de álcool, 47,4% eram alcoolistas, sendo a sua maioria classificada como alcoolista leve (59,3%). O grupo HFI foi composto por 54,5% pacientes do sexo masculino com uma média de idade de 57,3 anos. 90,9% dos pacientes eram idosos (>45 anos). A ocorrência das lesões foram em mucosa jugal (31,8%) e rebordo alveolar (25%). A expressão para p16Ink4a na LB foi fraca (41,8%) e para o grupo HFI, majoritariamente a expressão foi forte (68,2%). Real time PCR não detectou HPV-16 em nenhumas das amostras analisadas. Conclusão: Os achados em nosso estudo mostraram que a detecção de HPV de alto risco em tecido fresco, plasma e saliva para LB e HFI não se correlacionaram com a superexpressão da p16Ink4a.
Objective: To evaluate the expression of p16Ink4a by immunohistochemistry and the presence of HPV-16 by Real time PCR in fresh tissue, blood plasma, and saliva of patients with oral leukoplakia (OL) and inflammatory fibrous hyperplasia (IFH). Material and methods: Sixty-seven patients with the diagnosis of OL and 44 patients with the diagnosis of IFH were included in the study. Sociodemographic, clinicopathological, fresh tissue, blood, and saliva samples were collected and stored at - 80oC for molecular analysis. Paraffinized-embedded tissue was also used to perform the immunohistochemistry for p16Ink4a. Samples of biological material obtained from the patients were submitted to HPV-16 DNA detection by real time PCR, both with specific probes. Paraffinized-embedded tissue from the same patients were used to evaluate the expression of p16ink4a by immunohistochemistry. Results: Out of the 67 included OL patients, 55.2% were male with a mean age of 57.1 years. Elderly patients (> 45 years) comprised 86.6% of the sample. The prevalence of lesions was highest in the buccal mucosa (35.8%) followed by the mobile tongue (20.9%). 71.8% were smokers and the majority (47.8%) was classified as light smoker. Regarding alcohol consumption, 47.4% were alcoholics, most of whom were classified as light alcoholics (59.3%). The IFH group was composed of 54.5% of male patients with a mean age of 57.3 years. 90.9% of the sample were elderly patients (> 45 years). The highest occurrence of the lesions was in buccal mucosa (31.8%) and alveolar ridge (25%). The expression for p16Ink4a in LB was mostly weak (41.8%) and for the IFH group, the expression was mostly strong (68.2%). There was no positivity in any of the samples for HPV-16 by real time PCR. Conclusion: Our findings showed the HR-HPV detection in fresh tissue, plasma blood and saliva for OL and IFH does not correlate with p16Ink4a immunoexpression in paraffinized-embedded tissue.
Objective: To evaluate the expression of p16Ink4a by immunohistochemistry and the presence of HPV-16 by Real time PCR in fresh tissue, blood plasma, and saliva of patients with oral leukoplakia (OL) and inflammatory fibrous hyperplasia (IFH). Material and methods: Sixty-seven patients with the diagnosis of OL and 44 patients with the diagnosis of IFH were included in the study. Sociodemographic, clinicopathological, fresh tissue, blood, and saliva samples were collected and stored at - 80oC for molecular analysis. Paraffinized-embedded tissue was also used to perform the immunohistochemistry for p16Ink4a. Samples of biological material obtained from the patients were submitted to HPV-16 DNA detection by real time PCR, both with specific probes. Paraffinized-embedded tissue from the same patients were used to evaluate the expression of p16ink4a by immunohistochemistry. Results: Out of the 67 included OL patients, 55.2% were male with a mean age of 57.1 years. Elderly patients (> 45 years) comprised 86.6% of the sample. The prevalence of lesions was highest in the buccal mucosa (35.8%) followed by the mobile tongue (20.9%). 71.8% were smokers and the majority (47.8%) was classified as light smoker. Regarding alcohol consumption, 47.4% were alcoholics, most of whom were classified as light alcoholics (59.3%). The IFH group was composed of 54.5% of male patients with a mean age of 57.3 years. 90.9% of the sample were elderly patients (> 45 years). The highest occurrence of the lesions was in buccal mucosa (31.8%) and alveolar ridge (25%). The expression for p16Ink4a in LB was mostly weak (41.8%) and for the IFH group, the expression was mostly strong (68.2%). There was no positivity in any of the samples for HPV-16 by real time PCR. Conclusion: Our findings showed the HR-HPV detection in fresh tissue, plasma blood and saliva for OL and IFH does not correlate with p16Ink4a immunoexpression in paraffinized-embedded tissue.
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Palavras-chave
Imunohistoquímica, Leucoplasia, Papillomaviridae, Reação em cadeia da polimerase em tempo real, Immunohistochemistry