Effect of forced N-myc gene expression on src gene expression in neuroblastoma cells
Project/Area Number |
04670197
|
Research Category |
Grant-in-Aid for General Scientific Research (C)
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Allocation Type | Single-year Grants |
Research Field |
Experimental pathology
|
Research Institution | Chiba University |
Principal Investigator |
SIMIZU Bunsiti Chiba Univ., School of medicine, Professor, 医学部, 教授 (10072894)
|
Co-Investigator(Kenkyū-buntansha) |
SHIRASAWA Hiroshi Chiba Univ., School of medicine, Instructor, 医学部, 助手 (00216194)
|
Project Period (FY) |
1992 – 1993
|
Project Status |
Completed (Fiscal Year 1993)
|
Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1992: ¥1,500,000 (Direct Cost: ¥1,500,000)
|
Keywords | Neuroblastoma / N-myc / src |
Research Abstract |
NB69N which is a clonal subline of neuroblastoma cell line NB69, has a neuroblastic characteristic and an ability to differentiate by retinoic acid. S1 nuclease protection assay showed that NB69N expressed not only nonneuronal c-src but neuronal c-srcN1 and c-srcN2 mRNA.Because NB69N cell has a single copy of N-myc gene and does not express N-myc mRNA, we introduced N-myc expression vector into the NB69N cells by an electroporation method. We cloned 7 N-myc transfectants (NB69N41 - 47) and one control transfectant NB69NR.In 7 N-myc transfectants, the exogenous N-myc DNA was integrated in genomic DNA, and N-myc mRNA and protein products from the introduced N-myc were expressed. NB69N cells had no advantage of growth activity in a normal culture medium, but acquired a resistance to serum starvation. Furthermore, NB69N47 cells were able to form 6 times colonies to compare with NB69NR in soft agar. No difference of c-src mRNA splicing and expression levels was observed between NB69N47 and NB69NR by S1 nuclease protection assays. The forced expression of N-myc gene possibly conferred a malignant potential to the neuroblastoma cells independent of the c-src mRNA expression.
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Report
(3 results)
Research Products
(15 results)