Proteomic Identification of LASP-1 Down-regulation After RNAi Urokinase Silencing in Human Hepatocellular Carcinoma Cells
| Autor: | Francesca Miccichè, Alessandro Salvi, Italia Bongarzone, Sergio Barlati, Bruna Arici, Giuseppina De Petro |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Proteomics
Cancer Research Carcinoma Hepatocellular Difference gel electrophoresis Molecular Sequence Data Down-Regulation Biology LASP1 lcsh:RC254-282 Cytokeratin Western blot RNAi technology RNA interference Tumor Cells Cultured medicine Humans Gene silencing UK HCC Research Articles Adaptor Proteins Signal Transducing medicine.diagnostic_test Liver Neoplasms LIM Domain Proteins lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens Urokinase-Type Plasminogen Activator Urokinase receptor Cytoskeletal Proteins Cell culture Cancer research RNA Interference Ectopic expression |
| Zdroj: | Neoplasia: An International Journal for Oncology Research, Vol 11, Iss 2, Pp 207-219 (2009) |
| ISSN: | 1476-5586 |
| DOI: | 10.1593/neo.81076 |
| Popis: | In human hepatocellular carcinoma (HCC), the high expression of urokinase-type plasminogen activator (uPA) is an unfavorable prognostic factor and a therapeutic target. To identify the downstream effects of uPA silencing by RNA interference, we studied proteome modifications of uPA-inhibited SKHep1C3 cells, an HCC-derived cell line. The study with two-dimensional difference gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry showed Lim and SH3 protein 1 (LASP-1), cytokeratin 1 (CK-1), cytokeratin 10 (CK-10), and heterogeneous nuclear ribonucleoprotein H1 down-modulation after uPA inhibition. LASP-1, CK-1, and CK-10 are involved in cytoskeleton dynamics as heterogeneous nuclear ribonucleoprotein H1 takes part in the mRNA processing and stability. We first confirmed the proteomic data by Western blot and immunoflorescence and then explored the link between uPA and LASP-1. The ectopic expression of uPA and LASP-1 supported the proteomic results and showed that uPA up-regulation increased LASP-1 expression and that both were implicated in SKHep1C3 motility. siRNA LASP-1 inhibition showed that LASP-1 was involved in actin microfilaments organization of SKHep1C3 cells. The disruption of the actin microfilaments after LASP-1 depletion increased uPA secretion and SKHep1C3 motility. Our results would suggest the hypothesis that uPA and LASP-1 expression may be coordinated in HCC-derived cells. In summary, the proteomic identification of a set of uPA downstream proteins provides new insight into the function of uPA in HCC cells. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|