title: 2D difference gel electrophoresis analysis of different time points during the course of neoplastic transformation of human mammary epithelial cells.
abstract: Cell culture models of oncogenesis that use cellular reprogramming to generate a neoplastic cell from a normal cell provide one of the few opportunities to study the early stages of breast cancer development. Human mammary epithelial cells (HMECs) were induced to undergo a neoplastic transformation using defined genetic elements to generate transformed HMECs (THMECs). To identify proteins that displayed significantly different levels of abundance at three consecutive time points in oncogenesis over an 80 day period, protein extracts were analyzed by two-dimensional difference gel electrophoresis (2D-DIGE). Nine proteins were found to be significantly different in abundance: keratin 1, keratin 7, heat shock protein 4A-like, t-complex protein 1, stathmin, gelsolin, FK506 binding protein 5, ribosomal protein P0, and maspin. Keratin 7 and maspin displayed a linear down-regulation over 80 days. All of these proteins have been shown to be involved in the maintenance of a metastatic state including cytoskeletal modifications and motility. We conclude that, following neoplastic induction, THMECs display an early and progressive increase in metastatic potential. Further investigations into the function and regulatory mechanisms of these proteins will provide an unparalleled understanding of the initial states through which a breast cancer cell transitions following acquisition of the genetic abnormalities required for oncogenesis.
authors.affiliation: Department of Biology, UAB Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama 35294-1170, USA.
authors.forename: J Tyson
authors.initials: JT
authors.lastname: DeAngelis
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Li
authors.forename: Natalie
authors.initials: N
authors.lastname: Mitchell
authors.forename: Landon
authors.initials: L
authors.lastname: Wilson
authors.forename: Helen
authors.initials: H
authors.lastname: Kim
authors.forename: Trygve O
authors.initials: TO
authors.lastname: Tollefsbol
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism*; D002461:Cell Line, Transformed; D002471:Cell Transformation, Neoplastic / Q000378:metabolism*; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005260:Female; D018014:Gene Transfer Techniques; D006801:Humans; D042361:Mammary Glands, Human / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008954:Models, Biological; D009362:Neoplasm Metastasis; D020543:Proteome / Q000032:analysis / Q000378:metabolism; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015843:Serpins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014183:Transplantation, Heterologous
chemical_list: D020543:Proteome; C085344:SERPIN-B5; D015843:Serpins