title: High-throughput RNAi screening identifies a role for TNK1 in growth and survival of pancreatic cancer cells.
abstract: To identify novel targets in pancreatic cancer cells, we used high-throughput RNAi (HT-RNAi) to select genes that, when silenced, would decrease viability of pancreatic cancer cells. The HT-RNAi screen involved reverse transfecting the pancreatic cancer cell line BxPC3 with a siRNA library targeting 572 kinases. From replicate screens, approximately 32 kinases were designated as hits, of which 22 kinase targets were selected for confirmation and validation. One kinase identified as a hit from this screen was tyrosine kinase nonreceptor 1 (TNK1), a kinase previously identified as having tumor suppressor-like properties in embryonic stem cells. Silencing of TNK1 with siRNA showed reduced proliferation in a panel of pancreatic cancer cell lines. Furthermore, we showed that silencing of TNK1 led to increased apoptosis through a caspase-dependent pathway and that targeting TNK1 with siRNA can synergize with gemcitabine treatment. Despite previous reports that TNK1 affects Ras and NF-κB signaling, we did not find similar correlations with these pathways in pancreatic cancer cells. Our results suggest that TNK1 in pancreatic cancer cells does not possess the same tumor suppressor properties seen in embryonic cells but seems to be involved in growth and survival. The application of functional genomics by using HT-RNAi screens has allowed us to identify TNK1 as a growth-associated kinase in pancreatic cancer cells.
authors.affiliation: Clinical Translational Research Division, Translational Genomics Research Institute, Scottsdale, AZ 85259, USA.
authors.forename: Meredith C
authors.initials: MC
authors.lastname: Henderson
authors.forename: Irma M
authors.initials: IM
authors.lastname: Gonzales
authors.forename: Shilpi
authors.initials: S
authors.lastname: Arora
authors.forename: Ashish
authors.initials: A
authors.lastname: Choudhary
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Trent
authors.forename: Daniel D
authors.initials: DD
authors.lastname: Von Hoff
authors.forename: Spyro
authors.initials: S
authors.lastname: Mousses
authors.forename: David O
authors.initials: DO
authors.lastname: Azorsa
mesh_terms: D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005326:Fetal Proteins / Q000235:genetics / Q000502:physiology*; D020868:Gene Silencing; D057166:High-Throughput Screening Assays; D006801:Humans; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000473:pathology*; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics*; D000093542:Gemcitabine
chemical_list: D005326:Fetal Proteins; D034741:RNA, Small Interfering; D003841:Deoxycytidine; D011505:Protein-Tyrosine Kinases; C098672:TNK1 protein, human; D000093542:Gemcitabine