Functional analysis of polymorphic genes in oropharyngeal squamous cell carcinoma

• Grant number: 15/18039-6
• Support Opportunities: Regular Research Grants
• Duration: May 01, 2016 - December 31, 2018
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Carmen Silvia Passos Lima
• Grantee: Carmen Silvia Passos Lima
• Host Institution: Faculdade de Ciências Médicas (FCM). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated researchers: Albina Messias de Almeida Milani Altemani ; Gustavo Jacob Lourenço ; Manoela Marques Ortega

Abstract

Individual differences caused by single nucleotide polymorphisms (SNPs) in DNA damage repair, mechanisms of protein processing and transport, and mechanisms of intracellular transport genes indicate that the genetic constitution may predispose individuals to oropharyngeal (OP) squamous cell carcinoma (SCC) and influence patients prognosis. The roles of XPC c.2815A>C, ERP29 c.*293A>G and KIF13B c.*3163G>A SNPs in OPSCC are still unknown. Thus, the objectives of this study consists in: 1) to verify whether the distinct genotypes of XPC c.2815A>C, ERP29 c.*293A>G and KIF13B c.*3163G>A SNPs influence the OPSCC risk, clinical aspects of OPSCC patients and tumor characteristics; 2) to verify whether the distinct genotypes of XPC c.2815A>C, ERP29 c.*293A>G and KIF13B c.*3163G>A SNPs influence the gene expression in tumor cells, 3) to verify whether the distinct genotypes of XPC c.2815A>C, ERP29 c.*293A>G and KIF13B c.*3163G>A SNPs influence DNA repair ability, cell cycle and apoptosis in tumor cells; and 4) to verify whether the distinct genotypes of ERP29 c.*293A>G and KIF13B c.*3163G>A SNPs alter microRNAs miR-4421 and let-7e-3p expression, respectively. Genomic DNA of peripheral blood of 250 OPSCC patients and 250 controls will be evaluated to identify the genotypes of each SNP by real-time polymerase chain reaction (PCR), using TaqMan® (Applied Biosystems®) assays. The cell line of human pharynx SCC (FaDu) will be genetically modified to present homozygous wild-type and variant genotypes of XPC c.2815A>C, ERP29 c.*293A>G and KIF13B c.*3163G>A SNPs. Total RNA and protein will be obtained of peripheral blood from controls and tumor cells (ten with homozygous wild-type genotype, ten heterozygous and ten homozygous variant genotype of XPC c.2815A>C, ERP29 c.*293A>G and KIF13B c.*3163G>A SNPs) and from FaDu cell line (homozygous wild-type and variant genotypes of XPC c.2815A>C, ERP29 c.*293A>G and KIF13B c.*3163G>A SNPs). The genes and microRNAs expressions will be evaluated by quantitative PCR using specific primers and SYBR green® (Applied Biosystems®) and TaqMan® (Applied Biosystems®) assays, respectively. The protein quantification of samples will be evaluated by western blotting and immunohistochemistry methods. DNA repair, cell cycle and apoptosis will be evaluated by comet assay and flow cytometry method in FaDu cell line (homozygous wild-type and variant genotypes of XPC c.2815A>C, ERP29 c.*293A>G and KIF13B c.*3163G>A SNPs). The difference between groups will be analyzed by specific statistical tests. We believe that our results will contribute to define the SNPs roles in the SCCOP. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

CARRON, JULIANA; DALLA COSTA, ANA PAULA; RINCK-JUNIOR, JOSE AUGUSTO; MARIANO, FERNANDA VIVIANE; CARVALHO, BENILTON DE SA; PASSOS LIMA, CARMEN SILVIA; LOURENCO, GUSTAVO JACOB. Role of a genetic variation in the microRNA-4421 binding site of ERP29 regarding risk of oropharynx cancer and prognosis. SCIENTIFIC REPORTS, v. 10, n. 1, OCT 12 2020. (12/17182-1, 15/18039-6)