title: A novel cell line derived from pleomorphic adenoma expresses MMP2, MMP9, TIMP1, TIMP2, and shows numeric chromosomal anomalies.
abstract: Pleomorphic adenoma is the most common salivary gland neoplasm, and it can be locally invasive, despite its slow growth. This study aimed to establish a novel cell line (AP-1) derived from a human pleomorphic adenoma sample to better understand local invasiveness of this tumor. AP-1 cell line was characterized by cell growth analysis, expression of epithelial and myoepithelial markers by immunofluorescence, electron microscopy, 3D cell culture assays, cytogenetic features and transcriptomic study. Expression of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) was also analyzed by immunofluorescence and zymography. Furthermore, epithelial and myoepithelial markers, MMPs and TIMPs were studied in the tumor that originated the cell line. AP-1 cells showed neoplastic epithelial and myoepithelial markers, such as cytokeratins, vimentin, S100 protein and smooth-muscle actin. These molecules were also found in vivo, in the tumor that originated the cell line. MMPs and TIMPs were observed in vivo and in AP-1 cells. Growth curve showed that AP-1 exhibited a doubling time of 3.342 days. AP-1 cells grown inside Matrigel recapitulated tumor architecture. Different numerical and structural chromosomal anomalies were visualized in cytogenetic analysis. Transcriptomic analysis addressed expression of 7 target genes (VIM, TIMP2, MMP2, MMP9, TIMP1, ACTA2 e PLAG1). Results were compared to transcriptomic profile of non-neoplastic salivary gland cells (HSG). Only MMP9 was not expressed in both libraries, and VIM was expressed solely in AP-1 library. The major difference regarding gene expression level between AP-1 and HSG samples occurred for MMP2. This gene was 184 times more expressed in AP-1 cells. Our findings suggest that AP-1 cell line could be a useful model for further studies on pleomorphic adenoma biology.
authors.affiliation: Department of Oral and Maxillofacial Pathology, School of Dentistry, Federal University of Pará-UFPA, Belém, PA, Brazil.
authors.forename: Aline Semblano Carreira
authors.initials: AS
authors.lastname: Falcão
authors.affiliation: Department of Oral and Maxillofacial Pathology, School of Dentistry, Federal University of Pará-UFPA, Belém, PA, Brazil.
authors.forename: Maria Sueli da Silva
authors.initials: MS
authors.lastname: Kataoka
authors.affiliation: Laboratory of Molecular Biology, Evandro Chagas Institute, Belém, PA, Brazil.
authors.forename: Nélson Antonio Bailão
authors.initials: NA
authors.lastname: Ribeiro
authors.affiliation: Laboratory of Electron Microscopy, Evandro Chagas Institute, Belém, PA, Brazil.
authors.forename: José Antonio Picanço
authors.initials: JA
authors.lastname: Diniz
authors.affiliation: Department of Oral and Maxillofacial Pathology, School of Dentistry, Federal University of Pará-UFPA, Belém, PA, Brazil; Department of Oral and Maxillofacial Surgery, School of Dentistry, Universitary Center of Pará-CESUPA, Belém, Pará, Brazil.
authors.forename: Sérgio Melo
authors.initials: SM
authors.lastname: Alves
authors.affiliation: Department of Oral and Maxillofacial Pathology, School of Dentistry, Federal University of Pará-UFPA, Belém, PA, Brazil; Department of Oral and Maxillofacial Surgery, School of Dentistry, Universitary Center of Pará-CESUPA, Belém, Pará, Brazil.
authors.forename: André L Ribeiro
authors.initials: AL
authors.lastname: Ribeiro
authors.affiliation: Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
authors.forename: Adriane Sousa
authors.initials: AS
authors.lastname: de Siqueira
authors.affiliation: Institute of Biological Sciences, Federal University of Pará (Universidade Federal do Pará-UFPA), Belém, Pará, Brazil.
authors.forename: Artur Luiz
authors.initials: AL
authors.lastname: da Silva
authors.affiliation: Institute of Biological Sciences, Federal University of Pará (Universidade Federal do Pará-UFPA), Belém, Pará, Brazil.
authors.forename: Rommel Thiago Jucá
authors.initials: RT
authors.lastname: Ramos
authors.affiliation: Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
authors.forename: Vanessa M
authors.initials: VM
authors.lastname: Freitas
authors.affiliation: Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
authors.forename: Ruy G
authors.initials: RG
authors.lastname: Jaeger
authors.affiliation: Department of Oral and Maxillofacial Pathology, School of Dentistry, Federal University of Pará-UFPA, Belém, PA, Brazil; Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
authors.forename: João J V
authors.initials: JJ
authors.lastname: Pinheiro
mesh_terms: D008949:Adenoma, Pleomorphic / Q000473:pathology*; D000328:Adult; D014408:Biomarkers, Tumor / Q000378:metabolism; D045744:Cell Line, Tumor / Q000201:enzymology*; D002869:Chromosome Aberrations; D004252:DNA Mutational Analysis; D006801:Humans; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D012468:Salivary Gland Neoplasms / Q000473:pathology*; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism*; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000378:metabolism*; D059467:Transcriptome
chemical_list: D014408:Biomarkers, Tumor; C560822:TIMP1 protein, human; C560823:TIMP2 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D019716:Tissue Inhibitor of Metalloproteinase-2; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9