Downloaded from www.microbiologyresearch.org by IP: 54.70.40.11 On: Tue, 11 Dec 2018 12:57:28 Association of codon 72 polymorphism of p53 with the severity of cervical dysplasia, E6-T350G and HPV16 variant lineages in HPV16-infected women George D. Moschonas, 1 † Dimitris Tsakogiannis, 1, *† Konstantinos A. Lamprou, 1 Eirini Mastora, 1 Tilemachos G. Dimitriou, 1 Zaharoula Kyriakopoulou, 1 Christine Kottaridi, 2 Petros Karakitsos 2 and Panayotis Markoulatos 1 Abstract Purpose. Polymorphic variability in the tumour-suppressor protein p53 at codon 72 has a considerable impact on cervical cancer development. The present study clarified the association between p53 codon 72 genotypes and the risk of cervical disease in Greek patients. We also examined whether the presence of specific p53 genotypes in combination with HPV16 variants or E6 T350G sequence variation can modify an individual’s susceptibility to cervical disease. Methodology. The analysis of p53 genotypes was performed through PCR-RFLP. Sequence and phylogenetic tree analyses of the HPV16 E6 gene were also performed in order to identify HPV16 variants and T350G sequence variation. Results/Key findings. The outcomes of the present analysis revealed that women who are homozygous for the arg genotype are at a 4.17-fold higher risk of developing HPV16-associated HSIL+ (OR=4.17, 95 % CI:1.48–4.9, P=0.0049). Moreover, p53 arg/arg patients infected by an HPV16 prototype strain were associated with an increased risk of more severe lesions, while a significant relationship between the p53 arg/arg genotype in patients with T350G sequence variation and the risk of high- grade squamous intraepithelial lesions (HSILs) was revealed. Conclusion. The oncogenic potential of the virus is increased by the presence of the p53 arg/arg genotype in the Greek population in such a way that the specific protein interaction E6 (L83V)–p53 (Arg-72) can modify an individual’s susceptibility to cervical disease. INTRODUCTION Human papillomaviruses (HPVs) comprise a heterogeneous group of non-enveloped, capsid-enclosed, double-stranded DNA viruses that infect stratified epithelium, while HPV infections are characterized as mucosal or cutaneous [1, 2]. In general, it is thought that persistent infection with high-risk HPV genotypes is the leading cause for the development of precancerous and cervical cancer lesions, with the HPV16 genotype being detected in more than half of cervical cancer cases worldwide [3–5]. The increased prevalence of HPV16 DNA in cervical cancer triggered extensive investigation of the viral DNA sequence and HPV16 life cycle. In particular, sequence analysis of the long control region (LCR) and the E6 gene of HPV16 DNA revealed single nucleotide polymor- phisms that classify the viral genome into four major variant lineages and sub-lineages that are associated with specific geo- graphic and ethnic groups [6–11]. The HPV16 intratypic vari- ant lineages are described as (i) European–Asian (including the sub-lineages European and Asian); (ii) African type I; (iii) African type II; and (iv) Asian–American [11]. In recent decades several studies have investigated the distribution and oncogenic capacity of the nucleotide Received 22 February 2017; Accepted 19 July 2017 Author affiliations: 1 Department of Biochemistry and Biotechnology, Microbiology-Virology Laboratory, School of Health Sciences, University of Thessaly, Biopolis, 41500 Larissa, Greece; 2 Department of Cytopathology, National and Kapodistrian University of Athens, Medical School, ‘ATTIKON’ University Hospital, 1 Rimini, Haidari, Athens, 12462, Greece. *Correspondence: Dimitris Tsakogiannis, ditsakog@uth.gr or dtsakogiannis@gmail.com Keywords: HPV16; p53; T350G; HPV16 variants; HPV16 E6 gene; cervical cancer; p53 codon 72 Arg/Pro. Abbreviations: GTR + G, general time reversible plus gamma; GuSCN, guanidine thiocyanate; HPV, Human papillomavirus; HSIL, high-grade squa- mous intraepithelial lesion; LCR, long control region; LSIL, low-grade squamous intraepithelial lesion; MEGA, molecular evolutionary genetics analy- sis; MUSCLE, multiple sequence comparison by log-expectation; OR, odds ratio; RFLP, restriction fragment length polymorphism. †These authors contributed equally to this work. The GenBank/EMBL/DDBJ accession numbers for the E6-E7 sequences of HPV16 strains are KY564367–KY564398. In memory of Petros Karakitsos, an outstanding scientist and an excellent teacher. RESEARCH ARTICLE Moschonas et al., Journal of Medical Microbiology 2017;66:1358–1365 DOI 10.1099/jmm.0.000563 000563 ã 2017 The Authors 1358