In vitro RNA interference against beta-catenin inhibiting the proliferation of pediatric hepatic tumors
| Project/Area Number |
16591785
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pediatric surgery
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| Research Institution | Osaka University |
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Principal Investigator |
KUSAFUKA Takeshi Osaka University, Graduate School of Medicine, Assistant, 医学系研究科, 助手 (70263267)
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| Co-Investigator(Kenkyū-buntansha) |
YONEDA Akihiro Osaka University, Graduate School of Medicine, Assistant, 医学系研究科, 助手 (30372618)
FUKUZAWA Masahiro Osaka University, Graduate School of Medicine, Professor, 医学系研究科, 教授 (60165272)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2004: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | hepatoblastoma / HepG2 / Huh-6 / beta-catenin / RNA interference / pediatric |
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| Research Abstract |
To address beta-catenin's capacity in maintaining the malignant phenotype in established pediatric hepatoblastoma and hepatocellular carcinoma cell lines, HuH-6,and HepG2,harboring mutated and overexpressed beta-catenin, we carried out a series of in vitro analyses through a transfection of short interfering RNAs to generate a loss of function model. HuH-7,another hepatoblastoma cell line derived from a pediatric patient without a stabilizing mutation was used for comparison. RNA interference successfully manipulated the degradation of overexpressed beta-catenin. In all cell lines, beta-catenin mRNA was suppressed by 80-90 % after 48 h of transfection, and a reduction of its protein expression was demonstrated. In HuH-6 and HepG2,the pre-existing beta-catenin nuclear accumulation disappeared and reductions of beta-catenin downstream target genes, c-myc and cyclinD1,were also evidenced after treatment. The in vitro proliferation of both cell lines was transiently inhibited. In contrast, the suppression of beta-catenin in HuH-7 did not lead to a significant change in the expression of target genes or cellular proliferation. Our data indicate that beta-catenin can be considered a specific target for gene therapy in pediatric hepatic tumors with mutations and overexpression of this gene.
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Report
(3 results)
Research Products
(3 results)
