title: Fenretinide Perturbs Focal Adhesion Kinase in Premalignant and Malignant Human Oral Keratinocytes. Fenretinide's Chemopreventive Mechanisms Include ECM Interactions.
abstract: The membrane-associated protein, focal adhesion kinase (FAK), modulates cell-extracellular matrix interactions and also conveys prosurvival and proliferative signals. Notably, increased intraepithelial FAK levels accompany transformation of premalignant oral intraepithelial neoplasia (OIN) to oral squamous cell carcinoma (OSCC). OIN chemoprevention is a patient-centric, optimal strategy to prevent OSCC's comorbidities and mortality. The cancer chemopreventive and synthetic vitamin A derivative, fenretinide, has demonstrated protein-binding capacities, for example, mTOR- and retinol-binding protein interactions. These studies used a continuum of human oral keratinocytes (normal-HPV E6/E7-transduced-OSCC) to assess potential fenretinide-FAK drug protein interactions and functional consequences on cellular growth regulation and motility. Molecular modeling studies demonstrated that fenretinide has approximately 200-fold greater binding affinity relative to the natural ligand (ATP) at FAK's kinase domain. Fenretinide also shows intermediate binding at FAK's FERM domain and interacts at the ATP-binding site of the closest FAK analogue, PYK2. Fenretinide significantly suppressed proliferation via induction of apoptosis and G2-M cell-cycle blockade. Fenretinide-treated cells also demonstrated F-actin disruption, significant inhibition of both directed migration and invasion of a synthetic basement membrane, and decreased phosphorylation of growth-promoting kinases. A commercially available FAK inhibitor did not suppress cell invasion. Notably, although FAK's FERM domain directs cell invasion, FAK inhibitors target the kinase domain. In addition, FAK-specific siRNA-treated cells showed an intermediate cell migration capacity; data which suggest cocontribution of the established migrating-enhancing PYK2. Our data imply that fenretinide is uniquely capable of disrupting FAK's and PYK2's prosurvival and mobility-enhancing effects and further extend fenretinide's chemopreventive contributions beyond induction of apoptosis and differentiation.
authors.affiliation: Division of Biosciences, College of Dentistry, The Ohio State University, Columbus, Ohio.
authors.forename: Byungdo B
authors.initials: BB
authors.lastname: Han
authors.affiliation: Division of Oral Maxillofacial Pathology and Radiology, College of Dentistry, The Ohio State University, Columbus, Ohio.
authors.forename: Suyang
authors.initials: S
authors.lastname: Li
authors.affiliation: Division of Oral Maxillofacial Pathology and Radiology, College of Dentistry, The Ohio State University, Columbus, Ohio.
authors.forename: Meng
authors.initials: M
authors.lastname: Tong
authors.affiliation: Division of Biosciences, College of Dentistry, The Ohio State University, Columbus, Ohio.
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Holpuch
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio.
authors.forename: Richard
authors.initials: R
authors.lastname: Spinney
authors.affiliation: Division of Oral Maxillofacial Pathology and Radiology, College of Dentistry, The Ohio State University, Columbus, Ohio.
authors.forename: Daren
authors.initials: D
authors.lastname: Wang
authors.affiliation: Division of Oral Maxillofacial Pathology and Radiology, College of Dentistry, The Ohio State University, Columbus, Ohio.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Border
authors.affiliation: Division of Pharmaceutics and Pharmaceutical Chemistry, College of Pharmacy, The Ohio State University, Columbus, Ohio.
authors.forename: Zhongfa
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Division of Oral Maxillofacial Pathology and Radiology, College of Dentistry, The Ohio State University, Columbus, Ohio.
authors.forename: Sachin
authors.initials: S
authors.lastname: Sarode
authors.affiliation: Division of Oral Maxillofacial Pathology and Radiology, College of Dentistry, The Ohio State University, Columbus, Ohio.
authors.forename: Ping
authors.initials: P
authors.lastname: Pei
authors.affiliation: College of Pharmacy, University of Michigan, Ann Arbor, Michigan.
authors.forename: Steven P
authors.initials: SP
authors.lastname: Schwendeman
authors.affiliation: Division of Oral Maxillofacial Pathology and Radiology, College of Dentistry, The Ohio State University, Columbus, Ohio. The Ohio State University Comprehensive Cancer, Columbus, Ohio. mallery.1@osu.edu.
authors.forename: Susan R
authors.initials: SR
authors.lastname: Mallery
mesh_terms: D016588:Anticarcinogenic Agents / Q000494:pharmacology; D002294:Carcinoma, Squamous Cell / Q000473:pathology*; D002465:Cell Movement / Q000187:drug effects; D018890:Chemoprevention; D005109:Extracellular Matrix / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D017313:Fenretinide / Q000494:pharmacology*; D051417:Focal Adhesion Kinase 1 / Q000187:drug effects / Q000378:metabolism; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism*; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects* / Q000473:pathology*; D009061:Mouth Mucosa / Q000187:drug effects / Q000473:pathology; D009062:Mouth Neoplasms / Q000473:pathology*; D011230:Precancerous Conditions / Q000473:pathology*; D011485:Protein Binding / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured
chemical_list: D016588:Anticarcinogenic Agents; D017313:Fenretinide; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496050:PTK2 protein, human