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Ethanol Exposure Impacts Exosome Biogenesis of HeLa Cell-derived Exosomes

Graduate #25
Discipline: Biological Sciences
Subcategory: Microbiology/Immunology/Virology

Leandra B. Jones - Alabama State University
Co-Author(s): Jayde S. Price, Alabama State University, AL; Sparkle D. Williams, University of Alabama at Birmingham, AL; Alexandre Krendelchtchikov, University of Alabama at Birmingham, AL; Brian Sims University of Alabama at Birmingham, AL; and Qiana L. Matthews Alabama State University, AL



Exosomes are microvesicles that range from 30 – 100 nm in size. They can be found in various bodily fluids (breast milk, blood, semen, urine) and function mainly in cell-to-cell communication. Exosomes allow intracellular communication via DNA, RNA, and protein trafficking. External stressors such as alcohol have been known to alter the biogenesis and release of these vesicles. Currently, we are quantifying alcohol’s effects on exosome biogenesis at varying time points and concentrations in HeLa (cervical cancer) cells. HeLa cell were treated with 50 mM and 100 mM of Ethanol, respectively, for 24, 48 and 72 hours. Cell viability was observed at 72 hours post Ethanol treatment. HeLa cell viability was significantly decreased with treatments of 50 mM and 100 mM of Ethanol. After treatment, exosomes were purified from media via high-speed centrifugation. Exosome quantity was determined by the Lowry protein quantitation method. NanoSight technology was used to measure exosome diameter and exosome count after alcohol exposure. Enzyme-Linked Immunosorbent Assay was performed on exosomes to detect various exosome-specific proteins. The presence of exosomes was confirmed by the detection of various tetraspanins (CD9, CD63, CD81), as well as chaperone proteins (HSP90 Beta, HSP70, HSP60). Our findings show that alcohol-derived exosomes contain significant quantities of HSP60, HSP70, Fas, and Caspase-9 when compared to control- derived exosomes.

ERN LJones 10-13-17.docx

Funder Acknowledgement(s): Alabama State University

Faculty Advisor: Qiana Matthews, qmatthews@alasu.edu

Role: I performed majority of, actual laboratory work, ie. cell culture, cell viability, ELISA, SDS-Page, and Nanosight

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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