Is it safe to utilize in vitro reconstituted human oral epithelium? An oncogenetic pathway study.
Cell therapy is a therapeutic strategy used to replace or repair damaged tissue. The epithelium transplantation of cultivated keratinocytes has been applied to several modalities of reconstruction, like oral, urethra and ocular surface. Life and death signals work coordinately to ensure cellular quality control and the viability of an organism. The aim of this study is to verify that culture conditions did not induce genetic mutations through the analysis of the key genes: pAKT, Pten, p53 and MDM2 and investigate the presence of the related proteins in human oral keratinocytes obtained by primary culture and in vitro cultivated. Formalin fixed and paraffin embedded tissues from the oral cavity were utilized as control for normal expression of the related markers and two oral squamous cell carcinoma cell lines provided the expression pattern of the proposed markers in the event of cellular transformation. Akt, PTEN, p53 and MDM2 immunohistochemistry and Western-Blotting analyzes were performed. The results showed the expression levels and intracellular localizations of the four proteins evaluated. These analyzes confirmed that the produced in vitro epithelium is bio-compatible for its utilization as reconstruction and reparatory tissue, however further analyses and additional research on other biomarkers should be performed to analyse the long term engraftment of transplantable primary culture of oral keratinocytes and the long term resistance to cellular transformation.
Klingbeil
María Fátima Guarizo
MF
Department Oral Pathology, School of Dentistry, University of São Paulo, Avenida Professor Lineu Prestes, 2227, Cidade Universitária, São Paulo 05508-900, Brazil. fakling@usp.br
Mathor
Monica Beatriz
MB
Giudice
Fernanda Salgueiredo
FS
Yoshito
Daniele
D
dos Santos Pinto
Décio
D
D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000473:pathology; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000473:pathology*; D002478:Cells, Cultured; D004848:Epithelium / Q000473:pathology*; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D015603:Keratinocytes / Q000201:enzymology / Q000473:pathology / Q000637:transplantation; D009055:Mouth / Q000473:pathology*; D009062:Mouth Neoplasms / Q000473:pathology*; D051059:PTEN Phosphohydrolase / Q000378:metabolism; D021381:Protein Transport; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D051736:Proto-Oncogene Proteins c-mdm2 / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D016159:Tumor Suppressor Protein p53; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2; D051057:Proto-Oncogene Proteins c-akt; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human