Discovery of proteins associated with cervical cancer pathology

Doctoral Dissertation uoadl:2850669 87 Read counter

Τομέας Υγείας - Μητέρας - Παιδιού
Library of the School of Health Sciences
Deposit date:
Lygirou Vasiliki
Dissertation committee:
Παππά Καλλιόπη, Αναπληρώτρια Καθηγήτρια, Ιατρική Σχολή, ΕΚΠΑ
Ηλιόπουλος Αριστείδης, Καθηγητής, Ιατρική Σχολή, ΕΚΠΑ
Λουτράδης Δημήτριος, Καθηγητής, Ιατρική Σχολή, ΕΚΠΑ
Βάκας Παναγιώτης, Αναπληρωτής Καθηγητής, Ιατρική Σχολή, ΕΚΠΑ
Δασκαλάκης Γεώργιος, Αναπληρωτής Καθηγητής, Ιατρική Σχολή, ΕΚΠΑ
Γρηγοριάδης Θεμιστοκλής, Επίκουρος Καθηγητής, Ιατρική Σχολή, ΕΚΠΑ
Ρουμπελάκη Μαρία, Επίκουρη Καθηγήτρια, Ιατρική Σχολή, ΕΚΠΑ
Original Title:
Ανακάλυψη πρωτεϊνών σχετικών με την παθολογία του καρκίνου του τραχήλου της μήτρας
Translated title:
Discovery of proteins associated with cervical cancer pathology
Cervical cancer is the fourth most common and most lethal type of cancer in women, worldwide. Profylactic vaccination and regular screening have marked a decline on incidents, however the disease still accounts for a significant number of deaths and the available therapeutic approaches affect patients’ fertility. The purpose of this doctoral dissertation is the use of proteomic techniques in combination with bioinformatics and literature mining for the discovery of novel proteins involved in cervical cancer pathology with the aim of better understanding of the molecular mechanisms underlying the malignancies formation. To approach the goal, four cell lines, representative of the most common HPV infections (HeLa - HPV 18, SiHa - HPV 16), HPV-free cervical cancer (C33A) as well as normal cervical keratinocytes (HCK1T) were used. Analysis of the protein extracts was performed based on the GeLC-MS/MS protocol and provided a satisfactory number of protein identifications (2,500-3,500 proteins per cell line) with a very good reproducibility. The differentially expressed proteins that resulted from the comparison of each cervical cancer cell line with the normal keratinocytes were ~800-1,400 per comparison, and they provide useful information on the deregulation of signaling pathways and important molecules in cervical cancer. Bioinformatics analysis of the differentially expressed proteins validated the specificity of the identifications and the accuracy of the measurements as the deregulated molecular pathways are associated with the high metabolic demands and increased cell cycle that are known to occur in cancer cells. For the shortlisting of interesting proteins for further investigation, 105 differentially expressed proteins that followed the same expression trend in the three comparisons (HeLa/HCK1T, SiHa/CK1T και C33A/HCK1T) were chosen and extensively investigated in the literature. The proteins that were not studied in the context of cervical cancer before but have data on other types of cancers with similar expression trend as in the present study, are in total 21 and constitute the basis for further investigation and experiments. In addition, after a deeper investigation, 7 of these proteins were chosen (LIM domain and actin-binding protein 1, Importin subunit alpha-1, Serum paraoxonase/arylesterase 2, Elongation factor 1-alpha 2, Caprin-1, Mitochondrial import receptor subunit TOM34, Ras GTPase-activating protein-binding protein 1) for immunohistochemistry experiments on cervical specimens from healthy participants and patients with low and high grade squamous intraepithelial lesions and cervical cancer, as a next step to the present doctoral dissertation. Furthermore, functional investigation experiments for the proteins LIM domain and actin-binding protein 1, Importin subunit alpha-1 και Ras GTPase-activating protein-binding protein have been designed. In conclusion, the results of this doctoral dissertationprovide novel knowledge on the molecular mechanisms and the key regulators of cervical carcinogenesis and can be the basis for further investigations through systems biology approaches and contribute in the discovery of biomarkers and the identification of novel therapeutic targets.
Main subject category:
Health Sciences
Cervical cancer, Proteomics, Proteins, Liquid chromatography, Mass spectrometry
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