Alterations in the expression pattern of MTHFR, DHFR, TYMS, and SLC19A1 genes after treatment of laryngeal cancer cells with high and low doses of methotrexate

Autor(es)

Silva Galbiatti, Ana Livia ^[1] ; Castro, Rodrigo ^[1] ; Caldas, Heloisa Cristina ^[2] ; Padovani, Jr., Joao Armando ^[3] ; Pavarino, Erika Cristina ^[1] ; Goloni-Bertollo, Eny Maria ^[1]

Afiliação do(s) autor(es)

^[1] Sao Jose do Rio Preto Med Sch FAMERP, Genet & Mol Biol Res Unit UPGEM, Sao Jose Do Rio Preto, SP - Brazil

^[2] Sao Jose do Rio Preto Med Sch FAMERP, Lab Expt Immunol & Transplantat LITEX, Sao Jose Do Rio Preto, SP - Brazil

^[3] Sao Jose do Rio Preto Med Sch FAMERP, Otorhinolaryngol & Head & Neck Surg Dept, Sao Jose Do Rio Preto, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: TUMOR BIOLOGY; v. 34, n. 6, p. 3765-3771, DEC 2013.
• Citações Web of Science: 9
• Assunto(s): Expressão gênica Quimioterapia Neoplasias de cabeça e pescoço

Resumo

Inter-individual variations to methotrexate (MTX) outcome have been attributed to different expression profiles of genes related to folate metabolism. To elucidate the mechanisms of variations to MTX outcome, we investigated MTHFR, DHFR, TYMS, and SLC19A1 gene expression profiles by quantifying the mRNA level of the genes involved in folate metabolism to MTX response in laryngeal cancer cell line (HEP-2). For this, three different concentrations of MTX (0.25, 25, and 75 mu mol) were added separately in HEP-2 cell line for 24 h at 37 A degrees C. Apoptotis quantification was evaluated with fluorescein isothiocyanate-labeled Bcl-2 by flow cytometry. Real-time quantitative PCR technique was performed by quantification of gene expression with TaqManA (R) Gene Expression Assay. ANOVA and Bonferroni's post hoc tests were utilized for statistical analysis. The results showed that the numbers of apoptotic HEP-2 cells with 0.25, 25.0, and 75.0 mu mol of MTX were 14.57, 77.54, and 91.58 %, respectively. We found that the expression levels for MTHFR, DHFR, TYMS, and SLC19A1 genes were increased in cells with 75.0 mu mol of MTX (p < 0.05). Moreover, SLC19A1 gene presented lower expression in cells treated with 0.25 mu mol of MTX (p < 0.05). In conclusion, our data suggest that MTHFR, DHFR, TYMS, and SLC19A1 genes present increased expression after the highest application of MTX dose in laryngeal cancer cell line. Furthermore, SLC19A1 gene also presents decreased expression after the lowest application of MTX dose in laryngeal cancer cell line. Significant alterations of expression of these studied genes in cell culture model may give support for studies in clinical practice and predict interesting and often novel mechanisms of resistance of MTX chemotherapy. (AU)

• Processo FAPESP: 10/12930-4 - Avaliação de quimioterápicos, expressão gênica e quantificação de proteínas em carcinoma de cabeça e pescoço
• Beneficiário: Eny Maria Goloni Bertollo
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 10/12932-7 - Avaliação de quimioterápicos, expressão gênica e quantificação de proteínas em carcinoma de cabeça e pescoço
• Beneficiário: Ana Lívia Silva Galbiatti-Dias
• Modalidade de apoio: Bolsas no Brasil - Doutorado