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Analysis of the effects of the co-expression of E6 and E7 proteins from alfa and Beta-HPV in primary human keratinocytes

Grant number: 16/20327-2
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): January 01, 2017
Effective date (End): December 31, 2018
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal Investigator:Laura Cristina Sichero Vettorazzo
Grantee:Valéria Talpe Nunes
Host Institution: Instituto do Câncer do Estado de São Paulo Octavio Frias de Oliveira (ICESP). Coordenadoria de Serviços de Saúde (CSS). Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil


Infection by high risk oncogenic ±(Alpha)-HPV is the main etiologic factor for cervical cancer. Additionally, it has been suggested that some ²(Beta)-HPV types confer a higher risk regarding the development of non-melanoma skin tumors. Cases of co-infection by multiple HPV types have been reported, but it is not known as yet the functional effects of the interactions between the different HPVs types within the same infection. E6 and E7 proteins from ±- and ²-HPV genus, attain different strategies of association to cellular proteins to promote cellular proliferation and viral replication. Thus, the analysis of the expression of both proteins could elucidate the possible biological synergism or antagonism in the context of a co-infection. Thus, primary human keratinocytes will be co-transduced with E6 and E7 oncoproteins from HPV-18 (±-HPV genus) and HPV-38 or -49 (²-HPV genus); or will be independently transduced with the viral proteins of a single viral type, in order to examine the effects of these proteins in a situation of single infection and co-infection with HPV of different genera. Therefore, we will analyze: (1) the potential of primary human keratinocytes transduced with the viral oncoproteins to form colonies; (2) the potential of primary human keratinocytes transduced with the viral oncoproteins to inhibit differentiation induced by serum and calcium; (3) the expression levels of the cellular proteins p53, p21 and deltaNp73 by Western blot under the different conditions. The data generated are highly relevant to better understand the effects of multiple HPV types detection within the same anatomical site. (AU)

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