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Evaluation of chemotherapy, gene expression and proteins quantification in head and neck carcinoma


Chemotherapy with the methotrexate (MTX) and 5-fluorouracil (5-FU) antifolate drug in head and neck carcinoma may present acute toxicity and side effects and some tumors may show resistance to these agents. Furthermore, the cytotoxic effect is dependent on a competitive interaction with folate metabolism and therefore the high expression or decreased of certain genes this pathway have an important role in modulating the clinical toxicity and efficacy of these drugs. The objectives of this study are to evaluate the efficacy in vitro of chemotherapeutic MTX and 5-FU on FaDu cell line; quantify mRNA expression of proteins from genes DHFR, TS, MTHFR and RFC1 in cells treated with chemotherapy and to correlate with the gene expression. This will be used FaDu cell line (squamous cell carcinoma of the pharynx), after which cell culture will be given three different doses of each chemotherapy (MTX: 0.25 mM, 25 mM and 75 mM and 5-FU : 10 ng / ml, 50 ng / ml and 100 ng / ml). After this procedure, mRNA and proteins will be extracted for analysis of gene expression DHFR, TS, MTHFR and RFC1 involved in folate metabolism, which will be quantified by quantitative real time PCR and Western blotting techniques. It is expected, with the results of this proposal contribute to new therapeutic strategies for this disease, as prognostic factors with greater sensitivity and specificity can provide insight to new therapeutic strategies providing benefits to patients and new information regarding decision treatments (AU)

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Scientific publications (4)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
ZARA-LOPES, TAIRINE; SILVA GALBIATTI-DIAS, ANA LIVIA; URBANIN CASTANHOLE-NUNES, MARCIA M.; PADOVANI-JUNIOR, JOAO ARMANDO; MANIGLIA, JOSE VICTOR; PAVARINO, ERIKA CRISTINA; GOLONI-BERTOLLO, ENY MARIA. Polymorphisms in MTHFR, MTR, RFC1 and C beta S genes involved in folate metabolism and thyroid cancer: a case-control study. Archives of Medical Science, v. 15, n. 2, p. 522-530, . (12/14781-1, 10/12930-4)
ZARA-LOPES, T.; GIMENEZ-MARTINS, A. P. A.; NASCIMENTO-FILHO, C. H. V.; CASTANHOLE-NUNES, M. M. U.; GALBIATTI-DIAS, A. L. S.; PADOVANI-JUNIOR, J. A.; MANIGLIA, J. V.; FRANCISCO, J. L. E.; PAVARINO, E. C.; GOLONI-BERTOLLO, E. M.. Role of MTHFR C677T and MTR A2756G polymorphisms in thyroid and breast cancer development. Genetics and Molecular Research, v. 15, n. 2, . (10/12930-4, 12/14781-1)
SILVA GALBIATTI, ANA LIVIA; CALDAS, HELOISA CRISTINA; MANIGLIA, JOSE VICTOR; PAVARINO, ERIKA CRISTINA; GOLONI-BERTOLLO, ENY MARIA. Gene expression profile of 5-fluorouracil metabolic enzymes in laryngeal cancer cell line: Predictive parameters for response to 5-fluorouracil-based chemotherapy. BIOMEDICINE & PHARMACOTHERAPY, v. 68, n. 5, p. 515-519, . (10/12930-4, 10/12932-7)
SILVA GALBIATTI, ANA LIVIA; CASTRO, RODRIGO; CALDAS, HELOISA CRISTINA; PADOVANI, JR., JOAO ARMANDO; PAVARINO, ERIKA CRISTINA; GOLONI-BERTOLLO, ENY MARIA. 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. TUMOR BIOLOGY, v. 34, n. 6, p. 3765-3771, . (10/12930-4, 10/12932-7)

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