1Oncogenomic and Epigenetic Unit, Italian National Cancer Institute“Regina Elena”, Rome, Italy,2Genoma Group, Rome, Italy,3Otolaryngology Department, Italian National Cancer Institute“Regina Elena”, Rome, Italy,4Pathology Department, Italian National Cancer Institute“Regina Elena”, Rome, Italy Introduction: Head and neck squamous cell carcinoma (HNSCC) is characterized by a high incidence of relapse, which is the common cause of death in HNSCC patients. The identification of biomarkers supporting the manage-ment of HNSCC disease is still an unmet need in clinical oncology.
Meterials and Methods: To this end, we designed a mutational chip for next generation sequencing (NGS) analysis to detect mutations in tumor tissues and matched plasma of HNSCC patients. The analysis of the HNSCC TCGA cohort revealed that TP53 (72%), CDKN2A (22%) and FAT1 (24%) are the most frequently mutated genes in HNSCCs. Notably TP53 mutations associate with poor outcome. A cohort of 250 fresh frozen tissues from HNSCC patients has been collected. Specifically, for each patient three biopsies representing non-tumoral (resection margin), peritumor (histologically tumor-free tissue at≥1cm from the tumor) and tumor tissues were collected.
Results: This cohort has been challenged with a customized mutational chip for NGS analysis of mutations that includes the entire CDS of the TP53, CDKN2A and FAT1 genes. We found that the TP53 (743%) and CDKN2A (243%) mutation frequency in tumors of our cohort was similar to that of TGCA. While the frequency of FAT1 (38,6%) mutations was higher in our dataset. Many of the identified FAT1 mutations have not been annotated yet. Since we have also collected matched plasma at the surgery and during follow-up, our analysis is progressing toward the identification of mutations in ccf-DNA from patients.
Conclusions: This analysis is ongoing and the related data will be presented.
F. Ganci:None. F. Spinella: None. G. Cottone: None. E. Cotroneo: None. A. Nuccitelli: None. A. Biroccio: None. A. Sacconi: None. V. Manciocco: None. M. Pallocca: None. F. Rollo: None. R. Covello: None. M. Benevolo:None. G. Spriano: None. F. Fiorentino: None. G. Blandino:None.
P12.098B
Impact of cross-talk between laryngeal cancer cells and endothelial cells on cell migration and interactions
Z. B. Sari1, E. Yavuz2, T. Cora1
1Department of Medical Genetics, Selcuk University, Medical Faculty, Selcuklu, konya, Turkey,2Selcuk University Advanced
Technology Research and Application Center, Selcuklu, Konya, Turkey
Introduction:Endothelial cells constitute an important part of the tumor microenvironment (TME). However, the underlying mechanisms of their functions within the microenvironment of head and neck squamous cell carci-noma (HNSCC) remain poorly understood. Here we per-formed an in vitro non-contact co-cultivation system to analyze the influence of microenvironment in both tumor (laryngeal cancer cell line HEp-2) and healthy cells (endo-thelial cell line HUVEC cells).
Materials and Methods: We prepared conditioned medium (CM) from cell cultures for migration assay. CM has components secreted by cells such as specific cytokines, chemokines, growth factors. CM provides a co-culture microenvironments for the cells. We have used in vitro strach assay to examine cell migration and cell interactions. Photos were taken at intervals of 0, 3, 6, 9, 12 and 24 h. To assess the exact speed of cell migration, ImageJ software was used to measure the change in the cell-covered area over time, which is the characteristic parameter in migration assays.
Results: The wound healing assay showed that HEp-2 cells grown in CM from HUVEC have less capacity for migration and mobility compared with HEp-2 cells grown in the culture medium alone. HUVEC cells grown in CM from HEp-2 have less capacity for migration and mobility compared with HUVEC cells grown in the culture medium alone.
Conclusions: In vitro co-culture cell system for investi-gation of interaction between tumor cells and the tumor microenvironment was used. We believe that this study will contribute to our understanding of tumor microenvironment effect in head and neck carcinoma.
Z.B. Sari:None. E. Yavuz: None. T. Cora: None. P12.099C
Pathologies of helicases and premature ageing: study by derivation of induced pluripotent stem cells
V. Gatinois1, R. Desprat2, C. Corsini3, J. Puechberty4, J. B. Gaillard1, A. Schneider1, C. Lemattre1, T. Guignard1, J. M.
Lemaitre2, F. Pellestor1
1Unité de génétique chromosomique - Hôpital A. de Villeneuve, Montpellier, France,2Plateforme SAFE-iPS - Hôpital St Eloi, Montpellier, France,3Service d'Oncogénétique - Hôpital A. de Villeneuve, Montpellier, France,4Service de génétique médicale - Hôpital A. de Villeneuve, Montpellier, France Helicases process the double-stranded DNA dissociation. They are involved in replication, DNA repair and
