SET and sphingolipids in head and neck squamous cell carcinoma: signaling, targets and antiumoral therapy

Abstract

Despite the reached breakthrough in the treatment of some cancer types, the search for new therapeutic targets and antitumor therapy strategies to overcome tumor metastases, relapse, and resistance to therapy, is required. The present research proposal aims to identify evidences on the role of SET protein regarding sphingolipid signaling pathway, as well as the sphingolipids role, in head and neck squamous cell carcinoma (HNSCC). This approach can identify new therapeutic targets that may lead to new anticancer treatments. Several experimental approaches will be employed either in vitro or in vivo. We will perform studies in human cell lines, tumor samples, and serum of patients with cancer. In addition, different methods will be used to achieve the proposed goals, such as (i) sphingolipid analysis by mass spectrometry, (ii) post translational modifications of proteins by Western blotting, (iii) cell distribution of proteins by immunohistochemistry, immunofluorescence or Western blotting (subcellular fractionation), (iv) RNA and miRNAs quantification by real-time PCR, (v) DNA sequencing, (vi) RNA interference using either siRNA or shRNA, (vii) ectopic expression using vectors containing cDNAs of interest, (viii) flow cytometry for cell cycle analysis and characterization of cell types using antibodies, (ix) angiogenesis using HUVEC cells, and (x) cell viability, invasion and clonogenic assays. These studies, in addition to the tumor formation in nude mice using the xenograft model of human tumor cell lines and murine carcinoma cell lines in syngeneic mouse model, can both improve the knowledge on sphingolipid signaling in head and neck cancer and led to new mechanistic/therapeutic approaches for cancer. (AU)