The E6 and E7 proteins of beta3 human papillomavirus 49 can deregulate both cellular and extracellular vesicles-carried microRNAs
Abstract Background The β3 human papillomavirus (HPV)49 induces immortalization of primary keratinocytes through the action of E6 and E7 oncoproteins with an efficiency similar to alpha high risk (HR)-HPV16. Since HR-HPV oncoproteins are known to alter microRNA (miRNA) expression and extracellular v...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-06-01
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| Series: | Infectious Agents and Cancer |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13027-022-00445-z |
| _version_ | 1828813350566363136 |
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| author | Maria Vincenza Chiantore Marco Iuliano Roberta Maria Mongiovì Sankhadeep Dutta Massimo Tommasino Paola Di Bonito Luisa Accardi Giorgio Mangino Giovanna Romeo |
| author_facet | Maria Vincenza Chiantore Marco Iuliano Roberta Maria Mongiovì Sankhadeep Dutta Massimo Tommasino Paola Di Bonito Luisa Accardi Giorgio Mangino Giovanna Romeo |
| author_sort | Maria Vincenza Chiantore |
| collection | DOAJ |
| description | Abstract Background The β3 human papillomavirus (HPV)49 induces immortalization of primary keratinocytes through the action of E6 and E7 oncoproteins with an efficiency similar to alpha high risk (HR)-HPV16. Since HR-HPV oncoproteins are known to alter microRNA (miRNA) expression and extracellular vesicle (EV) production, we investigated the impact of HPV49 E6 and E7 proteins on miRNA profile and EV expression, and their involvement in the control of cell proliferation. Methods The miRNA expression was evaluated by a miRNA array and validated by RT-qPCR in primary human keratinocytes immortalized by β3 HPV49 (K49) or α9 HR-HPV16 (K16), and in EVs from K49 and K16. The modulation of miRNA target proteins was investigated by immunoblotting analyses. Results By comparing miRNA expression in K49 and K16 and the derived EVs, six miRNAs involved in HPV tumorigenesis were selected and validated. MiR-19a and -99a were found to be upregulated and miR-34a downregulated in both cell lines; miR-17 and -590-5p were upregulated in K49 and downmodulated in K16; miR-21 was downregulated only in K16. As for EV-carried miRNAs, the expression of miR-17, -19a, -21 and -99a was decreased and miR-34a was increased in K49 EVs. In K16 EVs, we revealed the same modulation of miR-19a, -34a, and -99a observed in producing cells, while miR-21 was upregulated. Cyclin D1, a common target of the selected miRNAs, was downmodulated in both cell lines, whereas cyclin-dependent kinase 4 was down-modulated in K49 but upregulated in K16. Conclusion These data suggest that E6 and E7 proteins of β3 HPV49 and α9 HR-HPV16 affect key factors of cell cycle control by indirect mechanisms based on miRNA modulation. |
| first_indexed | 2024-12-12T09:59:02Z |
| format | Article |
| id | doaj.art-74a3721619604b89b8f929549d2b597e |
| institution | Directory Open Access Journal |
| issn | 1750-9378 |
| language | English |
| last_indexed | 2024-12-12T09:59:02Z |
| publishDate | 2022-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Infectious Agents and Cancer |
| spelling | doaj.art-74a3721619604b89b8f929549d2b597e2022-12-22T00:28:02ZengBMCInfectious Agents and Cancer1750-93782022-06-0117111210.1186/s13027-022-00445-zThe E6 and E7 proteins of beta3 human papillomavirus 49 can deregulate both cellular and extracellular vesicles-carried microRNAsMaria Vincenza Chiantore0Marco Iuliano1Roberta Maria Mongiovì2Sankhadeep Dutta3Massimo Tommasino4Paola Di Bonito5Luisa Accardi6Giorgio Mangino7Giovanna Romeo8Department of Infectious Diseases, Istituto Superiore di SanitàDepartment of Medico-Surgical Sciences and Biotechnologies, Sapienza University of Rome - Polo PontinoDepartment of Medico-Surgical Sciences and Biotechnologies, Sapienza University of Rome - Polo PontinoDepartment of Oncogene Regulation, Chittaranjan National Cancer InstituteInfections and Cancer Biology Group, International Agency for Research on CancerDepartment of Infectious Diseases, Istituto Superiore di SanitàDepartment of Infectious Diseases, Istituto Superiore di SanitàDepartment of Medico-Surgical Sciences and Biotechnologies, Sapienza University of Rome - Polo PontinoDepartment of Medico-Surgical Sciences and Biotechnologies, Sapienza University of Rome - Polo PontinoAbstract Background The β3 human papillomavirus (HPV)49 induces immortalization of primary keratinocytes through the action of E6 and E7 oncoproteins with an efficiency similar to alpha high risk (HR)-HPV16. Since HR-HPV oncoproteins are known to alter microRNA (miRNA) expression and extracellular vesicle (EV) production, we investigated the impact of HPV49 E6 and E7 proteins on miRNA profile and EV expression, and their involvement in the control of cell proliferation. Methods The miRNA expression was evaluated by a miRNA array and validated by RT-qPCR in primary human keratinocytes immortalized by β3 HPV49 (K49) or α9 HR-HPV16 (K16), and in EVs from K49 and K16. The modulation of miRNA target proteins was investigated by immunoblotting analyses. Results By comparing miRNA expression in K49 and K16 and the derived EVs, six miRNAs involved in HPV tumorigenesis were selected and validated. MiR-19a and -99a were found to be upregulated and miR-34a downregulated in both cell lines; miR-17 and -590-5p were upregulated in K49 and downmodulated in K16; miR-21 was downregulated only in K16. As for EV-carried miRNAs, the expression of miR-17, -19a, -21 and -99a was decreased and miR-34a was increased in K49 EVs. In K16 EVs, we revealed the same modulation of miR-19a, -34a, and -99a observed in producing cells, while miR-21 was upregulated. Cyclin D1, a common target of the selected miRNAs, was downmodulated in both cell lines, whereas cyclin-dependent kinase 4 was down-modulated in K49 but upregulated in K16. Conclusion These data suggest that E6 and E7 proteins of β3 HPV49 and α9 HR-HPV16 affect key factors of cell cycle control by indirect mechanisms based on miRNA modulation.https://doi.org/10.1186/s13027-022-00445-zHuman papillomavirusmicroRNAsExtracellular vesiclesOncoproteinsHPV cancer |
| spellingShingle | Maria Vincenza Chiantore Marco Iuliano Roberta Maria Mongiovì Sankhadeep Dutta Massimo Tommasino Paola Di Bonito Luisa Accardi Giorgio Mangino Giovanna Romeo The E6 and E7 proteins of beta3 human papillomavirus 49 can deregulate both cellular and extracellular vesicles-carried microRNAs Infectious Agents and Cancer Human papillomavirus microRNAs Extracellular vesicles Oncoproteins HPV cancer |
| title | The E6 and E7 proteins of beta3 human papillomavirus 49 can deregulate both cellular and extracellular vesicles-carried microRNAs |
| title_full | The E6 and E7 proteins of beta3 human papillomavirus 49 can deregulate both cellular and extracellular vesicles-carried microRNAs |
| title_fullStr | The E6 and E7 proteins of beta3 human papillomavirus 49 can deregulate both cellular and extracellular vesicles-carried microRNAs |
| title_full_unstemmed | The E6 and E7 proteins of beta3 human papillomavirus 49 can deregulate both cellular and extracellular vesicles-carried microRNAs |
| title_short | The E6 and E7 proteins of beta3 human papillomavirus 49 can deregulate both cellular and extracellular vesicles-carried microRNAs |
| title_sort | e6 and e7 proteins of beta3 human papillomavirus 49 can deregulate both cellular and extracellular vesicles carried micrornas |
| topic | Human papillomavirus microRNAs Extracellular vesicles Oncoproteins HPV cancer |
| url | https://doi.org/10.1186/s13027-022-00445-z |
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