TY - JOUR
TI - Polymorphic variability in the exon 19 of the RB1 gene and its flanking intronic sequences in HPV16-associated precancerous lesions in the Greek population
AU - Tsakogiannis, D.
AU - Moschonas, G.D.
AU - Daskou, M.
AU - Stylianidou, Z.
AU - Kyriakopoulou, Z.
AU - Kottaridi, C.
AU - Dimitriou, T.G.
AU - Gartzonika, C.
AU - Markoulatos, P.
JO - Journal of Medical Microbiology
PY - 2018
VL - 67
TODO - 11
SP - 1638-1644
PB - Microbiology Society
SN - 0022-2615, 1473-5644
TODO - 10.1099/jmm.0.000843
TODO - amplicon;  Article;  clinical article;  exon;  female;  genetic variability;  Greece;  human;  Human papillomavirus type 16;  intron;  molecular cloning;  molecular evolution;  nucleotide sequence;  precancer;  priority journal;  rb1 gene;  restriction fragment length polymorphism;  sequence analysis;  single nucleotide polymorphism;  squamous intraepithelial lesion of the cervix;  uterine cervix dysplasia;  virus gene;  Bayes theorem;  case control study;  ethnology;  exon;  genetic polymorphism;  genetics;  genotype;  Human papillomavirus type 16;  intron;  isolation and purification;  papillomavirus infection;  polymerase chain reaction;  prospective study;  torticollis;  uterine cervix carcinoma in situ;  uterine cervix tumor;  virology, RB1 protein, human;  retinoblastoma binding protein;  tumor marker;  ubiquitin protein ligase;  virus DNA, Bayes Theorem;  Biomarkers, Tumor;  Case-Control Studies;  Cervical Intraepithelial Neoplasia;  DNA, Viral;  Evolution, Molecular;  Exons;  Female;  Genotype;  Greece;  Human papillomavirus 16;  Humans;  Introns;  Papillomavirus Infections;  Polymerase Chain Reaction;  Polymorphism, Genetic;  Polymorphism, Restriction Fragment Length;  Precancerous Conditions;  Prospective Studies;  Retinoblastoma Binding Proteins;  Torticollis;  Ubiquitin-Protein Ligases;  Uterine Cervical Neoplasms
TODO - Purpose. The tumour suppressor protein RB plays a decisive role in negative control of the cell cycle, inhibiting tumour development. The present analysis investigated the prevalence of the nucleotide polymorphism A153104G, which is located at intron 18 of the RB1 gene, and investigated the impact of the polymorphic variability in the exon 19 and its flanking intronic sequences on the severity of cervical disease in HPV16-positive Greek women. Methodology. The nucleotide polymorphism A153104G was detected by PCR-RFLP assay, while the amplicons were further subjected to cloning and sequencing. Moreover, molecular evolutionary analysis was performed using the maximumlikelihood (ML) and empirical Bayesian (EB) methods in order to evaluate the selective pressure acting on exon 19 of the RB1 gene. Results/Key findings. The A153104G nucleotide polymorphism was only detected in one control case. Moreover, sequence analysis of the amplicons revealed that the polymorphic variability in the RB1 gene increased with the severity of the cervical dysplasia. The link between the observed polymorphic variability and the progress of cervical disease was reflected in the molecular evolutionary analysis that was performed on the exon 19 of the RB1 gene, since negative selective pressure was acting upon exon 19 in the control and low-grade squamous intraepithelial lesion (LSIL) cervical samples, while positive selective pressure was acting upon exon 19 in the high-grade squamous intraepithelial lesion (HSIL) specimens. Conclusions. The A153104G nucleotide polymorphism did not emerge as a potential biomarker for the development of precancerous lesions in the Greek patients, while the accumulation of sequence variations in RB1 gene might influence patients’ susceptibility towards the progression of cervical neoplasia. © 2018 The Authors.
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