Oxidative Stress in HPV-Driven Viral Carcinogenesis: Redox Proteomics Analysis of HPV-16 Dysplastic and Neoplastic Tissues

Genital infection by high risk Human Papillomavirus (HR-HPV), although recognized as the main etio-pathogenetic factor of cervical cancer, is not per se sufficient to induce tumour development. Oxidative stress (OS) represents an interesting and under-explored candidate as a promoting factor in HPV-initiated carcinogenesis. To gain insight into the role of OS in cervical cancer, HPV-16 positive tissues were collected from patients with invasive squamous cervical carcinoma, from patients with High Grade dysplastic HPV lesions and from patients with no clinical evidence of HPV lesions. After virological characterization, modulation of proteins involved in the redox status regulation was investigated. ERp57 and GST were sharply elevated in dysplastic and neoplastic tissues. TrxR2 peaked in dysplastic samples while iNOS was progressively reduced in dysplastic and neoplastic samples. By redox proteomic approach, five proteins were found to have increased levels of carbonyls in dysplastic samples respect to controls namely: cytokeratin 6, actin, cornulin, retinal dehydrogenase and GAPDH. In carcinoma samples the peptidyl-prolyl cis-trans isomerase A, ERp57, serpin B3, Annexin 2 and GAPDH were found less oxidized than in dysplastic tissues. HPV16 neoplastic progression seems associated with increased oxidant environment. In dysplastic tissues the oxidative modification of DNA and proteins involved in cell morphogenesis and terminal differentiation may provide the conditions for the neoplastic progression. Conversely cancer tissues seem to attain an improved control on oxidative damage as shown by the selective reduction of carbonyl adducts on key detoxifying/pro-survival proteins

Oxidative Stress in HPV-Driven Viral Carcinogenesis: Redox Proteomics Analysis of HPV-16 Dysplastic and Neoplastic Tissues

Genital infection by high risk Human Papillomavirus (HR-HPV), although recognized as the main etio-pathogenetic factor of cervical cancer, is not per se sufficient to induce tumour development. Oxidative stress (OS) represents an interesting and under-explored candidate as a promoting factor in HPV-initiated carcinogenesis. To gain insight into the role of OS in cervical cancer, HPV-16 positive tissues were collected from patients with invasive squamous cervical carcinoma, from patients with High Grade dysplastic HPV lesions and from patients with no clinical evidence of HPV lesions. After virological characterization, modulation of proteins involved in the redox status regulation was investigated. ERp57 and GST were sharply elevated in dysplastic and neoplastic tissues. TrxR2 peaked in dysplastic samples while iNOS was progressively reduced in dysplastic and neoplastic samples. By redox proteomic approach, five proteins were found to have increased levels of carbonyls in dysplastic samples respect to controls namely: cytokeratin 6, actin, cornulin, retinal dehydrogenase and GAPDH. In carcinoma samples the peptidyl-prolyl cis-trans isomerase A, ERp57, serpin B3, Annexin 2 and GAPDH were found less oxidized than in dysplastic tissues. HPV16 neoplastic progression seems associated with increased oxidant environment. In dysplastic tissues the oxidative modification of DNA and proteins involved in cell morphogenesis and terminal differentiation may provide the conditions for the neoplastic progression. Conversely cancer tissues seem to attain an improved control on oxidative damage as shown by the selective reduction of carbonyl adducts on key detoxifying/pro-survival proteins

Genital Human Papillomavirus infection and genotype prevalence among Albanian women. A cross sectional study.

International audience“High Risk” HPV types have different geographical distribution and evidence suggests their respective prevalence may vary in different areas and regions. Accurate description of High Risk HPV circulation is a key feature for the rational design of prevention and screening campaigns. A cross-sectional, virological study was conducted on adult Albanian women living either in the Tirana area or in the Duress prefecture. Clinical and gynaecological evaluations were done according to current standard criteria. HPV detection and typing were carried out by a combined MY09/MY11 and GP5+/GP6+ PCR followed by direct sequencing of generated amplicons. Virological data could be obtained from 402 out of 452 patients enrolled between January 2004 and December 2007. Sixtyone patients (15.1% of the cohort) were found to be infected with a genital HPV. As expected viral prevalence was higher among women younger than 30years of age (25.2%) in comparison to those aged 30 or older (13.6%). HPV 16 was found to be the most frequent type (41% of cases), followed by HPV 53, (roughly 7.2%) HPV 31 (5.8%) and HPV 18 (4.3%). HPV 81 and HPV 84, were the most prevalent low risk types detected with prevalences of 11.6 and 5.8 %. No difference was noted in any type specific prevalence between young and mature women. The circulation of HPV types is far more complex than assumed generally. The detailed knowledge of HPV type circulating patterns in specific local geographical areas is essential for appropriate implementation of screening, prevention and surveillance campaigns

Oxidative Stress in HPV-Driven Viral Carcinogenesis: Redox Proteomics Analysis of HPV-16 Dysplastic and Neoplastic Tissues

Genital infection by high risk Human Papillomavirus (HR-HPV), although recognized as the main etio-pathogenetic factor of cervical cancer, is not per se sufficient to induce tumour development. Oxidative stress (OS) represents an interesting and under-explored candidate as a promoting factor in HPV-initiated carcinogenesis. To gain insight into the role of OS in cervical cancer, HPV-16 positive tissues were collected from patients with invasive squamous cervical carcinoma, from patients with High Grade dysplastic HPV lesions and from patients with no clinical evidence of HPV lesions. After virological characterization, modulation of proteins involved in the redox status regulation was investigated. ERp57 and GST were sharply elevated in dysplastic and neoplastic tissues. TrxR2 peaked in dysplastic samples while iNOS was progressively reduced in dysplastic and neoplastic samples. By redox proteomic approach, five proteins were found to have increased levels of carbonyls in dysplastic samples respect to controls namely: cytokeratin 6, actin, cornulin, retinal dehydrogenase and GAPDH. In carcinoma samples the peptidyl-prolyl cis-trans isomerase A, ERp57, serpin B3, Annexin 2 and GAPDH were found less oxidized than in dysplastic tissues. HPV16 neoplastic progression seems associated with increased oxidant environment. In dysplastic tissues the oxidative modification of DNA and proteins involved in cell morphogenesis and terminal differentiation may provide the conditions for the neoplastic progression. Conversely cancer tissues seem to attain an improved control on oxidative damage as shown by the selective reduction of carbonyl adducts on key detoxifying/prosurvival proteins

Involvement of Oxidative Stress in Occurrence of Relapses in Multiple Sclerosis: The Spectrum of Oxidatively Modified Serum Proteins Detected by Proteomics and Redox Proteomics Analysis

<div><p>Multiple sclerosis (MS) is an autoimmune inflammatory demyelinating disease of the central nervous system. Several evidences suggest that MS can be considered a multi-factorial disease in which both genetics and environmental factors are involved. Among proposed candidates, growing results support the involvement of oxidative stress (OS) in MS pathology. The aim of this study was to investigate the role of OS in event of exacerbations in MS on serum of relapsing-remitting (RR-MS) patients, either in relapsing or remitting phase, with respect to serum from healthy subjects. We applied proteomics and redox proteomics approaches to identify differently expressed and oxidatively modified proteins in the low-abundant serum protein fraction. Among differently expressed proteins ceruloplasmin, antithrombin III, clusterin, apolipoprotein E, and complement C3, were up-regulated in MS patients compared with healthy controls. Further by redox proteomics, vitamin D-binding protein showed a progressive trend of oxidation from remission to relapse, respect with controls. Similarly, the increase of oxidation of apolipoprotein A-IV confirmed that levels of OS are elevated with the progression of the disease. Our findings support the involvement of OS in MS and suggest that dysfunction of target proteins occurs upon oxidative damage and correlates with the pathology.</p></div

Levels of protein carbonyls.

<p>Protein carbonyls (PC) levels in the serum of control (CTR) and multiple sclerosis patients: remitting (REM) and relapsing (REL). Serum samples were assayed for PC by slot blot analysis. Error bars indicate SD for 6 samples per group. Densitometric values shown are given as raw values.</p

Demographic and clinical characteristic of multiple sclerosis patients.

<p>EDSS = Expanded Disability Status Scale; Dis Dur = Disease duration; MSSS = Multiple sclerosis severity Score; MFIS = FatFatigue Severity Scale; BDI = Beck Depression Inventory; PASAT = Paced Auditory Serial Addition Test.</p><p>Re Reported values are ± SD.</p

Functions of Identified Proteins Differently Expressed and Oxidized.

<p>Functions of Identified Proteins Differently Expressed and Oxidized.</p

Apolipoprotein A-IV oxidation fold.

<p>On the left side of the figure, an enlarged area of 2D blots from CTR and REL groups (A), and from REM and REL (B),are shown corresponding to the Apolipoprotein A-IV (Apo A-IV) spot. Apo A-IV spot identified as differently oxidized in the matching between REL vs CTR, and REL vs REM is labeled. 3D density graphs are elaborated by PD-Quest from Apo A-IV spot on 2D blot. On the right side of the figure the histogram reports oxidation fold for Apo A-IV.</p

Spectra obtained by MALDI-ToF MS analysis related to spots of vitamin D-binding protein (DBP).

<p>DBP C-terminal region spectrum (upper panel) and major coverage DBP spectrum (lower panel).</p