1. Presence of HPV DNA in extracellular vesicles from HeLa cells and cervical samples
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Efraín Garrido, Sergio R. Aguilar-Ruiz, Javier Gaytan-Cervantes, Minerva Mata-Rocha, María de Los Ángeles Romero-Tlalolini, Pedro Chavez-Olmos, Carolina Gonzalez-Torres, Honorio Torres-Aguilar, Ruth Monserrat Rodríguez-Hernández, Juan Manuel Mejía-Aranguré, and Conrado Robles-Vázquez more...
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0301 basic medicine ,Microbiology (medical) ,030106 microbiology ,Cervix Uteri ,Genome ,Virus ,DNA sequencing ,HeLa ,Extracellular Vesicles ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,medicine ,Humans ,030212 general & internal medicine ,Cervix ,Cervical cancer ,biology ,Papillomavirus Infections ,Oncogene Proteins, Viral ,medicine.disease ,biology.organism_classification ,Molecular biology ,Microvesicles ,medicine.anatomical_structure ,chemistry ,DNA, Viral ,Female ,DNA ,HeLa Cells - Abstract
Introduction The main cause of cervical cancer is an infection of keratinocytes in the basal layer of the stratified epithelium of the cervix by human papillomavirus (HPV). Other than in cervical samples, HPV DNA has been found in serum and other fluids but its origin is unclear. Extracellular vesicles (EV) could be a conveyance of viral DNA given their emerging role in cellular communication. The content of EV derived from cervical cells has not been properly explored and it is not known whether or not they contain HPV DNA. Methods We evaluated the DNA content of exosomes purified from cultures of HeLa cells by Next Generation Sequencing (NGS) and confirmed its presence by PCR. The presence of HPV DNA was also evaluated by PCR and NGS in EV from HPV-positive cervical samples without apparent lesion or with LSIL. Results We detected the integrated form of viral-DNA in exosomes from HeLa cells by NGS and confirmed its presence by PCR. The search for HPV sequences in EV obtained from cervical exudate samples without apparent lesion or with LSIL, where we expected to find the viral genome as an episome, indicated that HPV DNA, including the E6 and E7 oncogenes, is present in these EV. Conclusion HPV DNA, including the viral oncogenes E6/E7, is found in exosomes regardless of the integration status of the virus in the infected cell. more...
- Published
- 2020
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