siRNA-Mediated β-Catenin Knockdown in Human Hepatoma Cells Results in Decreased Growth and Survival
β-Catenin, the chief oncogenic component of the canonical Wnt pathway, is known to be involved in a variety of cancers, including hepatocellular carcinoma (HCC). Although the mechanism of β-catenin activation in HCC is multifactorial, it is indisputably implicated at various stages of hepatocarcinog...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2007-11-01
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| Series: | Neoplasia: An International Journal for Oncology Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1476558607800236 |
| _version_ | 1811265700111581184 |
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| author | Gang Zeng Udayan Apte Benjamin Cieply Sucha Singh Satdarshan P.S. Monga |
| author_facet | Gang Zeng Udayan Apte Benjamin Cieply Sucha Singh Satdarshan P.S. Monga |
| author_sort | Gang Zeng |
| collection | DOAJ |
| description | β-Catenin, the chief oncogenic component of the canonical Wnt pathway, is known to be involved in a variety of cancers, including hepatocellular carcinoma (HCC). Although the mechanism of β-catenin activation in HCC is multifactorial, it is indisputably implicated at various stages of hepatocarcinogenesis, making it an attractive therapeutic target. Here we investigate the effect of small interfering RNA-mediated β-catenin knockdown on the growth and survival of human hepatoma cell lines with (HepG2) and without (Hep3B) β-catenin mutations. Transfection of HepG2 and Hep3B cells with human β-catenin (CTNNBl) small interfering RNA resulted in a significant β-catenin decrease, as confirmed by Western blot analyses and immunofluorescence, also leading to decreased expression of known target genes such as cyclin D1 and glutamine synthetase. The decrease in β-catenin activity was confirmed by TOPflash reporter luciferase assay. The functional impact of diminished β-catenin was exhibited as temporal decrease in tumor cell viability by the MTT assay. A concomitant decrease in tumor cell proliferation was also evident with [3H]thymidine incorporation and verified with soft agar assays. Thus, β-catenin is essential for the survival and growth of hepatoma cells independent of mutations in the β-catenin gene and provide a proof of principle for the significance of the therapeutic inhibition of β-catenin in HCC. |
| first_indexed | 2024-04-12T20:27:36Z |
| format | Article |
| id | doaj.art-c1a4ed6d325f45f69bcbfd931529cb75 |
| institution | Directory Open Access Journal |
| issn | 1476-5586 1522-8002 |
| language | English |
| last_indexed | 2024-04-12T20:27:36Z |
| publishDate | 2007-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Neoplasia: An International Journal for Oncology Research |
| spelling | doaj.art-c1a4ed6d325f45f69bcbfd931529cb752022-12-22T03:17:49ZengElsevierNeoplasia: An International Journal for Oncology Research1476-55861522-80022007-11-0191195195910.1593/neo.07469siRNA-Mediated β-Catenin Knockdown in Human Hepatoma Cells Results in Decreased Growth and SurvivalGang Zeng0Udayan Apte1Benjamin Cieply2Sucha Singh3Satdarshan P.S. Monga4Departments of Pathology, University of Pittsburgh, SOM, Pittsburgh, PA 15213. USADepartments of Pathology, University of Pittsburgh, SOM, Pittsburgh, PA 15213. USADepartments of Pathology, University of Pittsburgh, SOM, Pittsburgh, PA 15213. USADepartments of Pathology, University of Pittsburgh, SOM, Pittsburgh, PA 15213. USADepartments of Pathology, University of Pittsburgh, SOM, Pittsburgh, PA 15213. USAβ-Catenin, the chief oncogenic component of the canonical Wnt pathway, is known to be involved in a variety of cancers, including hepatocellular carcinoma (HCC). Although the mechanism of β-catenin activation in HCC is multifactorial, it is indisputably implicated at various stages of hepatocarcinogenesis, making it an attractive therapeutic target. Here we investigate the effect of small interfering RNA-mediated β-catenin knockdown on the growth and survival of human hepatoma cell lines with (HepG2) and without (Hep3B) β-catenin mutations. Transfection of HepG2 and Hep3B cells with human β-catenin (CTNNBl) small interfering RNA resulted in a significant β-catenin decrease, as confirmed by Western blot analyses and immunofluorescence, also leading to decreased expression of known target genes such as cyclin D1 and glutamine synthetase. The decrease in β-catenin activity was confirmed by TOPflash reporter luciferase assay. The functional impact of diminished β-catenin was exhibited as temporal decrease in tumor cell viability by the MTT assay. A concomitant decrease in tumor cell proliferation was also evident with [3H]thymidine incorporation and verified with soft agar assays. Thus, β-catenin is essential for the survival and growth of hepatoma cells independent of mutations in the β-catenin gene and provide a proof of principle for the significance of the therapeutic inhibition of β-catenin in HCC.http://www.sciencedirect.com/science/article/pii/S1476558607800236Liver cancerWnt/β-catenindevelopmenttreatmentregeneration |
| spellingShingle | Gang Zeng Udayan Apte Benjamin Cieply Sucha Singh Satdarshan P.S. Monga siRNA-Mediated β-Catenin Knockdown in Human Hepatoma Cells Results in Decreased Growth and Survival Neoplasia: An International Journal for Oncology Research Liver cancer Wnt/β-catenin development treatment regeneration |
| title | siRNA-Mediated β-Catenin Knockdown in Human Hepatoma Cells Results in Decreased Growth and Survival |
| title_full | siRNA-Mediated β-Catenin Knockdown in Human Hepatoma Cells Results in Decreased Growth and Survival |
| title_fullStr | siRNA-Mediated β-Catenin Knockdown in Human Hepatoma Cells Results in Decreased Growth and Survival |
| title_full_unstemmed | siRNA-Mediated β-Catenin Knockdown in Human Hepatoma Cells Results in Decreased Growth and Survival |
| title_short | siRNA-Mediated β-Catenin Knockdown in Human Hepatoma Cells Results in Decreased Growth and Survival |
| title_sort | sirna mediated β catenin knockdown in human hepatoma cells results in decreased growth and survival |
| topic | Liver cancer Wnt/β-catenin development treatment regeneration |
| url | http://www.sciencedirect.com/science/article/pii/S1476558607800236 |
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