TIMP3 and CCNA1 hypermethylation in HNSCC is associated with an increased incidence of second primary tumors

Background: Hypermethylation in the promoter regions is associated with the suppression of gene expression and has been considered a potential molecular marker for several tumor types, including head and neck squamous cell carcinomas (HNSCC).Methods: To evaluate the gene hypermethylation profile as a prognostic marker, this retrospective study used a QMSP approach to determine the methylation status of 19 genes in 70 HNSCC patients.Results: the methylation profile analysis of primary HNSCC revealed that genes CCNA1, DAPK, MGMT, TIMP3 and SFRP1 were frequently hypermethylated, with high specificity and sensitivity. TIMP3 and CCNA1 hypermethylation was significantly associated with lower rates of second primary tumor-free survival (p = 0.007 and p = 0.001; log-rank test, respectively).Conclusion: This study, for the first time, presents CCNA1 and TIMP3 hypermethylation as a helpful tool to identify HNSCC subjects at risk of developing second primary carcinomas.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Dept Biol Sci, Lab Mol Canc Biol, BR-04039020 São Paulo, BrazilBarretos Canc Hosp, Stat & Epidemiol Ctr, BR-14784400 Barretos, BrazilAC Camargo Hosp, Dept Head & Neck Surg, BR-01509010 São Paulo, BrazilBarretos Canc Hosp, Dept Head & Neck Surg, BR-14784400 Barretos, BrazilDuke NUS Grad Med Sch, Canc & Stem Cell Biol Program, Singapore 169857, SingaporeUniversidade Federal de São Paulo, Dept Biol Sci, Lab Mol Canc Biol, BR-04039020 São Paulo, BrazilWeb of Scienc

Clinical significance of molecular alterations in histologically negative surgical margins of head and neck cancer patients

The development of locoregional recurrence is the main reason for treatment failure in head and neck squamous cell carcinomas (HNSCC) and the remaining of tumor cells in surgical margins is associated with recurrence. Surgical margins are considered negative based on histologic assessment of the pathological specimen. However, this method lacks sensitivity in identifying cells that already started malignant transformation but have not yet developed a pathologic phenotype. We investigated the usefulness of assessing the expression of PTHLH, EPCAM, MMP9, LGALS1 and MET for the detection of molecular alterations in histologically negative surgical margins and determine the correlation of these tumor-related alterations with clinical and prognostic parameters. Differential gene expression was determined by quantitative RT-PCR analyses in normal mucosa, HNSCC and negative margin samples. Thirty-eight percent of the histologically negative surgical margins examined were margin-positive for overexpression of at least one of the genes evaluated. Moreover, MMP9 and PTHLH overexpression in the surgical margins was associated with the development of second primary tumors (p = 0.002) and lower rates of local control (log rank test p = 0.022; HR = 4.186; p = 0.035), respectively. These findings demonstrate that the overexpression of tumor-related genes in histologically negative surgical margins is a frequent event. the use of qRT-PCR may be an useful tool in detecting actually negative HNSCC surgical margins and the overexpression of specific genes in these margins could be helpful in the identification of patients with a higher risk of developing second primary tumors and local recurrences, thus aiding the surgeon in the delineation of the HNSCC resection extent and helping in the planning of adjuvant therapy. (C) 2011 Elsevier B.V. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Lab Mol Canc Biol, Dept Biol Sci, BR-04039032 São Paulo, BrazilHosp Canc AC Camargo, Fundacao Antonio Prudente, Dept Head & Neck Surg, BR-01509010 São Paulo, BrazilHosp Canc AC Camargo, Fundacao Antonio Prudente, Dept Pathol, BR-01509010 São Paulo, BrazilHosp Canc Barretos, Dept Head & Neck Surg, Fundacao PIO 12, BR-14784400 Barretos, SP, BrazilUniversidade Federal de São Paulo, Lab Mol Canc Biol, Dept Biol Sci, BR-04039032 São Paulo, BrazilFAPESP: 2007/56245-0FAPESP: 2008/58460-9CNPq: 302360/2008-5Web of Scienc

Accuracy of microRNAs as markers for the detection of neck lymph node metastases in patients with head and neck squamous cell carcinoma

Background: the presence of metastatic disease in cervical lymph nodes of head and neck squamous cell carcinoma (HNSCC) patients is a very important determinant in therapy choice and prognosis, with great impact in overall survival. Frequently, routine lymph node staging cannot detect occult metastases and the post-surgical histologic evaluation of resected lymph nodes is not sensitive in detecting small metastatic deposits. Molecular markers based on tissue-specific microRNA expression are alternative accurate diagnostic markers. Herein, we evaluated the feasibility of using the expression of microRNAs to detect metastatic cells in formalin-fixed paraffin-embedded (FFPE) lymph nodes and in fine-needle aspiration (FNA) biopsies of HNSCC patients.Methods: An initial screening compared the expression of 667 microRNAs in a discovery set comprised by metastatic and non-metastatic lymph nodes from HNSCC patients. the most differentially expressed microRNAs were validated by qRT-PCR in two independent cohorts: i) 48 FFPE lymph node samples, and ii) 113 FNA lymph node biopsies. the accuracy of the markers in identifying metastatic samples was assessed through the analysis of sensitivity, specificity, accuracy, negative predictive value, positive predictive value, and area under the curve values.Results: Seven microRNAs highly expressed in metastatic lymph nodes from the discovery set were validated in FFPE lymph node samples. MiR-203 and miR-205 identified all metastatic samples, regardless of the size of the metastatic deposit. Additionally, these markers also showed high accuracy when FNA samples were examined.Conclusions: the high accuracy of miR-203 and miR-205 warrant these microRNAs as diagnostic markers of neck metastases in HNSCC. These can be evaluated in entire lymph nodes and in FNA biopsies collected at different time-points such as pre-treatment samples, intraoperative sentinel node biopsy, and during patient follow-up. These markers can be useful in a clinical setting in the management of HNSCC patients from initial disease staging and therapy planning to patient surveillance.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Dept Biol Sci, Lab Canc Mol Biol, BR-04039032 São Paulo, SP, BrazilBarretos Canc Hosp, Mol Oncol Res Ctr, BR-14784400 Barretos, SP, BrazilBarretos Canc Hosp, Dept Pathol, BR-14784400 Barretos, SP, BrazilBarretos Canc Hosp, Dept Head & Neck Surg, BR-14784400 Barretos, SP, BrazilDuke NUS Grad Med Sch, Canc Stem Cell Biol Program, Singapore 169857, SingaporeUniversidade Federal de São Paulo, Dept Biol Sci, Lab Canc Mol Biol, BR-04039032 São Paulo, SP, BrazilFAPESP: 2012/14837-7Web of Scienc

The complete genome sequence of Chromobacterium violaceum reveals remarkable and exploitable bacterial adaptability

Chromobacterium violaceum is one of millions of species of free-living microorganisms that populate the soil and water in the extant areas of tropical biodiversity around the world. Its complete genome sequence reveals (i) extensive alternative pathways for energy generation, (ii) ≈500 ORFs for transport-related proteins, (iii) complex and extensive systems for stress adaptation and motility, and (iv) wide-spread utilization of quorum sensing for control of inducible systems, all of which underpin the versatility and adaptability of the organism. The genome also contains extensive but incomplete arrays of ORFs coding for proteins associated with mammalian pathogenicity, possibly involved in the occasional but often fatal cases of human C. violaceum infection. There is, in addition, a series of previously unknown but important enzymes and secondary metabolites including paraquat-inducible proteins, drug and heavy-metal-resistance proteins, multiple chitinases, and proteins for the detoxification of xenobiotics that may have biotechnological applications

Clonagem e caracterização do fator de transativação opaco 2 de Coix lacryma-jobi

Orientador: Adilson LeiteDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de BiologiaMestradoGenetica de PlantasMestre em Ciência

O ativador transcricional opaco2 : analise de seu promotor e efeito da fosforilação do seu dominio bZIP na interação proteina : DNA

Orientador: Adilson LeiteTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaDoutoradoGeneticaDoutor em Ciências Biológica

Identification of markers for the presence of lymph nodes metastasis in patients with oral squamous cell carcinomas

Universidade Federal de São Paulo, São Paulo, BrazilHosp Canc Barretos, Barretos, BrazilUniversidade Federal de São Paulo, São Paulo, BrazilWeb of Scienc

Evaluation of the methylation profile of exfoliated cell samples from patients with head and neck squamous cell carcinoma

BackgroundSilencing of tumor suppressor genes plays a vital role in head and neck carcinogenesis. the purposes of this study were to determine the methylation profile of exfoliated tumors cells collected from patients with head and neck squamous cell carcinoma (HNSCC) and to evaluate its prognostic significance.MethodsThe methylation profile and level of a 20-gene panel were evaluated by quantitative methylation-specific polymerase chain reaction (qMSP) in exfoliated tumor cell samples from 96 patients with HNSCC.ResultsCCNA1 (60.4%), DCC (54.2%), and TIMP3 (35.4%) were frequently methylated in these samples. Patients with exfoliated tumors cells positive for DCC methylation showed a trend toward a lower local recurrence-free survival.ConclusionThese findings indicate that a low invasive method could be used to access the methylation profile of exfoliated cells from patients with HNSCC. Moreover, our data provide evidence that hypermethylation of DCC could be useful as prognostic indicator for this malignancy. (c) 2013 Wiley Periodicals, Inc. Head Neck 36: 631-637, 2014Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Dept Ciencias Biol, Lab Biol Mol Canc, São Paulo, BrazilHosp AC Camargo Fund Antonio Prudente, Dept Cirurgia Cabeca & Pescoco, São Paulo, BrazilHosp Canc Barretos, Dept Cirurgia Cabeca & Pescoco, Barretos, BrazilUniversidade Federal de São Paulo, Dept Ciencias Biol, Lab Biol Mol Canc, São Paulo, BrazilFAPESP: 2008/58460-9FAPESP: 2005/02580-8FAPESP: 07/56471-0FAPESP: 07/56245-0CNPq: 313181/2009-8CNPq: 302360/2008-5Web of Scienc