Background: The β3 human papillomavirus (HPV)49 induces immortalization of primary keratinocytes through the action of E6 and E7 oncoproteins with an efciency 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 profle 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 afect key factors of cell cycle control by indirect mechanisms based on miRNA modulation.

The E6 and E7 proteins of beta3 human papillomavirus 49 can deregulate both cellular and extracellular vesicles-carried microRNAs / Vincenza Chiantore, Maria; Iuliano, Marco; Mongiovi, Roberta Maria; Dutta, Sankhadeep; Tommasino, Massimo; Di Bonito, Paola; Accardi, Luisa; Mangino, Giorgio; Romeo, Giovanna. - In: INFECTIOUS AGENTS AND CANCER. - ISSN 1750-9378. - 17:(2022), pp. 1-12. [10.1186/s13027-022-00445-z]

The E6 and E7 proteins of beta3 human papillomavirus 49 can deregulate both cellular and extracellular vesicles-carried microRNAs

Marco Iuliano
Co-primo
;
Roberta Maria Mongiovì
Secondo
;
Giorgio Mangino
Penultimo
;
Giovanna Romeo
Ultimo
2022

Abstract

Background: The β3 human papillomavirus (HPV)49 induces immortalization of primary keratinocytes through the action of E6 and E7 oncoproteins with an efciency 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 profle 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 afect key factors of cell cycle control by indirect mechanisms based on miRNA modulation.
2022
human papillomavirus; microRNAs; extracellular vesicles; oncoproteins; HPV cancer
01 Pubblicazione su rivista::01a Articolo in rivista
The E6 and E7 proteins of beta3 human papillomavirus 49 can deregulate both cellular and extracellular vesicles-carried microRNAs / Vincenza Chiantore, Maria; Iuliano, Marco; Mongiovi, Roberta Maria; Dutta, Sankhadeep; Tommasino, Massimo; Di Bonito, Paola; Accardi, Luisa; Mangino, Giorgio; Romeo, Giovanna. - In: INFECTIOUS AGENTS AND CANCER. - ISSN 1750-9378. - 17:(2022), pp. 1-12. [10.1186/s13027-022-00445-z]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1646028
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