title: GRK2 negatively regulates IGF-1R signaling pathway and cyclins' expression in HepG2 cells.
abstract: G protein coupled receptor kinase 2 (GRK2) plays a central role in the regulation of a variety of important signaling pathways. Alternation of GRK2 protein level and activity casts profound effects on cell physiological functions and causes diseases such as heart failure, rheumatoid arthritis, and obesity. We have previously reported that overexpression of GRK2 has an inhibitory role in cancer cell growth. To further examine the role of GRK2 in cancer, in this study, we investigated the effects of reduced protein level of GRK2 on insulin-like growth factor 1 receptor (IGF-1R) signaling pathway in human hepatocellular carcinoma (HCC) HepG2 cells. We created a GRK2 knockdown cell line using a lentiviral vector mediated expression of GRK2 specific short hairpin RNA (shRNA). Under IGF-1 stimulation, HepG2 cells with reduced level of GRK2 showed elevated total IGF-1R protein expression as well as tyrosine phosphorylation of receptor. In addition, HepG2 cells with reduced level of GRK2 also demonstrated increased tyrosine phosphorylation of IRS1 at the residue 612 and increased phosphorylation of Akt, indicating a stronger activation of IGF-1R signaling pathway. However, HepG2 cells with reduced level of GRK2 did not display any growth advantage in culture as compared with the scramble control cells. We further detected that reduced level of GRK2 induced a small cell cycle arrest at G2/M phase by enhancing the expression of cyclin A, B1, and E. Our results indicate that GRK2 has contrasting roles on HepG2 cell growth by negatively regulating the IGF-1R signaling pathway and cyclins' expression.
authors: Division of Transplantation, Department of Surgery, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA. wei-zhengyu@cooperhealth.edu
authors: Zhengyu
authors: Z
authors: Wei
authors: Reginald
authors: R
authors: Hurtt
authors: Tina
authors: T
authors: Gu
authors: Adam S
authors: AS
authors: Bodzin
authors: Walter J
authors: WJ
authors: Koch
authors: Cataldo
authors: C
authors: Doria
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D059447:Cell Cycle Checkpoints / Q000235:genetics; D019925:Cyclin A / Q000235:genetics*; D056744:Cyclin B1 / Q000235:genetics*; D019927:Cyclin E / Q000378:metabolism*; D054769:G-Protein-Coupled Receptor Kinase 2 / Q000235:genetics / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D056945:Hep G2 Cells; D006801:Humans; D055504:Insulin Receptor Substrate Proteins / Q000235:genetics; D017526:Receptor, IGF Type 1 / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: C534699:CCNB1 protein, human; D019925:Cyclin A; D056744:Cyclin B1; D019927:Cyclin E; C526292:IRS1 protein, human; D055504:Insulin Receptor Substrate Proteins; D017526:Receptor, IGF Type 1; C496567:GRK2 protein, human; D054769:G-Protein-Coupled Receptor Kinase 2