title: Cervical Cancer Cell Line Secretome Highlights the Roles of Transforming Growth Factor-Beta-Induced Protein ig-h3, Peroxiredoxin-2, and NRF2 on Cervical Carcinogenesis.
abstract: Cancer cells acquire unique secretome compositions that contribute to tumor development and metastasis. The aim of our study was to elucidate the biological processes involved in cervical cancer, by performing a proteomic analysis of the secretome from the following informative cervical cell lines: SiHa (HPV16+), HeLa (HPV18+), C33A (HPV-), and HCK1T (normal). Proteins were analyzed by 2D gel electrophoresis coupled to MALDI-TOF-MS. Enrichment of secreted proteins with characteristic profiles for each cell line was followed by the identification of differentially expressed proteins. Particularly, transforming growth factor-beta-induced protein ig-h3 (Beta ig-h3) and peroxiredoxin-2 (PRDX2) overexpression in the secretome of cancer cell lines was detected and confirmed by Western blot. Bioinformatics analysis identified the transcription factor NRF2 as a regulator of differentially expressed proteins in the cervical cancer secretome. NRF2 levels were measured by both Western blot and Multiple Reaction Monitoring (MRM) in the total cell extract of the four cell lines. NRF2 was upregulated in SiHa and C33A compared to HCK1T. In conclusion, the secreted proteins identified in cervical cancer cell lines indicate that aberrant NRF2-mediated oxidative stress response (OSR) is a prominent feature of cervical carcinogenesis.
authors.affiliation: Biotechnology Division, Biomedical Research Foundation, Academy of Athens (BRFAA), Athens, Greece; Laboratory of Biology, University of Athens School of Medicine, Athens, Greece.
authors.forename: Georgia
authors.identifier: 0000-0002-4175-9019
authors.initials: G
authors.lastname: Kontostathi
authors.affiliation: Biotechnology Division, Biomedical Research Foundation, Academy of Athens (BRFAA), Athens, Greece.
authors.forename: Jerome
authors.initials: J
authors.lastname: Zoidakis
authors.affiliation: Biotechnology Division, Biomedical Research Foundation, Academy of Athens (BRFAA), Athens, Greece.
authors.forename: Manousos
authors.initials: M
authors.lastname: Makridakis
authors.affiliation: Biotechnology Division, Biomedical Research Foundation, Academy of Athens (BRFAA), Athens, Greece; Laboratory of Biology, University of Athens School of Medicine, Athens, Greece.
authors.forename: Vasiliki
authors.initials: V
authors.lastname: Lygirou
authors.affiliation: Biotechnology Division, Biomedical Research Foundation, Academy of Athens (BRFAA), Athens, Greece.
authors.forename: George
authors.initials: G
authors.lastname: Mermelekas
authors.affiliation: Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institute of Cardiovascular and Metabolic Disease, Toulouse, France; Université Toulouse III Paul-Sabatier, Toulouse, France.
authors.forename: Theofilos
authors.identifier: 0000-0003-1565-3875
authors.initials: T
authors.lastname: Papadopoulos
authors.affiliation: Biotechnology Division, Biomedical Research Foundation, Academy of Athens (BRFAA), Athens, Greece.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Vougas
authors.affiliation: Department of Chemistry, Division of Organic Chemistry, Biochemistry and Natural Products, University of Patras, 26504 Rion, Greece.
authors.forename: Alexios
authors.identifier: 0000-0001-5068-6459
authors.initials: A
authors.lastname: Vlamis-Gardikas
authors.affiliation: First Department of Obstetrics and Gynecology, University of Athens School of Medicine, Alexandra Hospital, Athens, Greece.
authors.forename: Peter
authors.initials: P
authors.lastname: Drakakis
authors.affiliation: First Department of Obstetrics and Gynecology, University of Athens School of Medicine, Alexandra Hospital, Athens, Greece.
authors.forename: Dimitrios
authors.identifier: 0000-0002-8686-594X
authors.initials: D
authors.lastname: Loutradis
authors.affiliation: Biotechnology Division, Biomedical Research Foundation, Academy of Athens (BRFAA), Athens, Greece.
authors.forename: Antonia
authors.identifier: 0000-0003-3284-5713
authors.initials: A
authors.lastname: Vlahou
authors.affiliation: Laboratory of Biology, University of Athens School of Medicine, Athens, Greece; Laboratory of Cell and Gene Therapy, Biomedical Research Foundation, Academy of Athens, Athens, Greece.
authors.forename: Nicholas P
authors.identifier: 0000-0001-7130-5192
authors.initials: NP
authors.lastname: Anagnou
authors.affiliation: First Department of Obstetrics and Gynecology, University of Athens School of Medicine, Alexandra Hospital, Athens, Greece; Laboratory of Cell and Gene Therapy, Biomedical Research Foundation, Academy of Athens, Athens, Greece.
authors.forename: Kalliopi I
authors.identifier: 0000-0002-1061-7405
authors.initials: KI
authors.lastname: Pappa
mesh_terms: D000465:Algorithms; D063646:Carcinogenesis; D045744:Cell Line, Tumor; D019295:Computational Biology; D015180:Electrophoresis, Gel, Two-Dimensional; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006367:HeLa Cells; D052162:Human papillomavirus 16; D006801:Humans; D051267:NF-E2-Related Factor 2 / Q000378:metabolism*; D030361:Papillomavirus Infections / Q000150:complications; D010455:Peptides / Q000737:chemistry; D054464:Peroxiredoxins / Q000378:metabolism*; D040901:Proteomics; D015398:Signal Transduction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D002583:Uterine Cervical Neoplasms / Q000378:metabolism*; D000099222:betaIG-H3 Protein
chemical_list: D016326:Extracellular Matrix Proteins; D051267:NF-E2-Related Factor 2; D010455:Peptides; D054464:Peroxiredoxins; D016212:Transforming Growth Factor beta; D000099222:betaIG-H3 Protein; C495635:NFE2L2 protein, human; C086065:PRDX2 protein, human