Lse S – t) and Sonneborn Stiftelse.Background: Infection with human papilloma virus (HPV) is an vital pathological issue in head and neck squamous cell carcinoma (HNSCC). Two categories of HNSCC might be distinguished when it comes to HPV status: HPV(+) and HPV(-). HNSCCs differ from each other in respect to their biology and response to therapy. Exosomes are produced by all living cells and mediate intercellular communication. Their protein profiles resemble these of parent cells. Exosomes interact with and reprogram functions of human immune cells. The aim of this study was to examine protein profiles of tumour cell-derived exosomes by mass spectrometry for the presence of proteins which could interact with immune cells and modulate their functions. Approaches: We studied protein profiles of exosomes released by cells of three HNSCC HPV(+) cell lines: SCC-2, SCC-47, SCC-90 and two HNSCC HPV(-) cell lines: PCI-13 and PCI-30. Exosomes had been isolated from tumour cell supernatants by min-size exclusion chromatography (mini-SEC). The isolated exosomes had been assessed for: (1) morphology and size by transmission electron microscopy; (two) number of vesicles by q-Nano; and (3) the protein content material. Molecular profiles have been determined making use of high-resolution tandem mass spectrometry (LC-MS/MS) method. The outcomes had been confirmed employing on-bead flow cytometry method. Benefits: Exosomes originating from HPV(+) and HPV(-) cancer cells had the identical size (3050 nm) and morphology. Having said that, only HPV(+) exosomes contained the following proteins: E6/E7, Rb and survivin, though HPV(-) exosomes had been unfavorable for cyclin D1 and had low levels of p53. Application of high-resolution mass spectrometry enabled detection of CD47 and CD276 receptor proteins detected only in exosomes originating from HPV(+) cells. Summary/Conclusion: As each of those proteins play important roles in HDAC6 Inhibitor Storage & Stability exosome interactions with immune cells, the information recommend that HPV(+) cancers modulate the host immune program differently than HPV(-) cancers. Funding: The analysis was financed in component by the Polish National Science Centre project no. [2013/11/B/NZ7/01512].LBT03.Proteomic analysis of exosomes released from irradiated head and neck cancer cells Agata Abramowicz1; Mateusz Smolarz1; Lukasz Marczak2; Piotr Widlak1; Monika PietrowskaMaria Sklodowska-Curie Institute – Oncology Center, Gliwice Branch, Gliwice, Poland; 2Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznan, PolandLBT03.Proteome of exosomes released by HPV(+) and HPV(-) head and neck cancer cells Monika Pietrowska1; Lukasz Marczak2; Agata Abramowicz1; Marta Gawin1; Sonja Funk3; Priyanka Sharma4; Piotr Widlak1; Theresa L. WhitesideBackground: Head and neck squamous cell carcinoma could be the sixth most typical cancer worldwide with a poor prognosis. Deeper understanding of resistance mechanisms induced in cancer cells for the duration of radiotherapy may contribute to improvement of HNSCC curability. We think that exosomes reported as essential players in intercellular communication may possibly play a important function in response to radiation along with other genotoxic agents employed in cancer remedy. Procedures: UM-SCC6 cells were irradiated with doses of two, 4, and 8 Gy and cell culture supernatant was collected after 24 h. ExosomeThursday, 03 Maysamples were purified by differential centrifugation and filtration (0.22 ), then supernatant was concentrated and lastly separated with SEC. Collected CaMK II Activator Compound fractions had been assessed by immunodetection of tetraspanin markers,.
