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Biomedical subjects

F Hiragami

Publications and source records attributed to F Hiragami.

5 recordsLinked to original sources

cAMP and calcium ionophore induce outgrowth of neuronal processes in PC12 mutant cells in which nerve growth factor-induced outgrowth of neuronal processes is impaired.

During continuous culturing, PC12 cells are subject to spontaneous mutations. We obtained PC12m3 cells, clone cells in which outgrowth of neuronal processes (dendrites and axons) under the condition of nerve growth factor (NGF) treatment was highly stimulated by various inducers, such as cyclic adenosine monophosphate (cAMP), calcium ionophore, steroid and high osmolarity. The number of cells with neuronal processes in the presence of cAMP was approximately twenty-fold greater than PC12 parental cells and other PC12 mutant cells. In PC12m3 cells, NGF-induced outgrowth of neuronal processes was reduced by cytotoxic solanine, whereas the effect of NGF was unaffected by hyaluronic acid. In PC12m3 cells, various inducers of neurite outgrowth, such as cAMP, calcium ionophore and high osmolarity, activated mitogen activated protein (MAP) kinase, whereas solanine and hyaluronic acid did not cause any significant activation of MAP kinase. However, PC12m3 cells, in which NGF-induced outgrowth of neuronal processes were impaired, had strong NGF-induced MAP kinase activity as PC12 parental cells had. These findings suggest that cAMP, calcium influx and high osmolarity induce outgrowth of neuronal processes in PC12m3 cells through activation of the downstream target of MAP kinase or through a novel pathway independent of NGF activation.

Animals↗

Morphological alteration of X-ray induced partially transformed human cells by transfection with a small c-myc DNA sequence.

During attempts to transform a normal human fibroblast strain (GM730) by X-irradiation, we obtained a partially transformed cell strain (GM730pt) which demonstrates several aspects of the transformed phenotype including morphological changes, increased saturation density, growth in soft agar, and focus formation in long-term cultures. When GM730pt cells were transfected with the feline c-myc gene, morphology of the cells changed dramatically following seven days of expression. Transfection of other plasmid DNAs or oncogenes such as pUC8, pSV2neo, src, sis, and H-ras had little or no effects on the phenotype of GM730pt cells. On the other hand, a gel purified, small fragment of c-myc DNA had a complete cell alteration activity. Furthermore, Bal 31 deletion and M13 sequencing experiments showed that the alteration seen in GM730pt cells is delimited to a 24 nucleotide stretch (active myc element) from the second intron of the feline c-myc gene that contains a T-rich sequence.

Agar↗

SV40LT highly mutates and immortalizes two fibroblast strains from patients with Wilms' tumor.

In order to analyze in detail the process of immortalization of human cells, SV40LT was introduced into two chromosome 11p- fibroblast strains from Wilms' tumor patients. Both fibroblasts, hereafter referred to as CM1 and CM2, displayed the mutant phenotype in the crisis stage of cellular aging. In comparison to a control fibroblast, the density of the CM1 strain was abnormally high while the crisis period of the CM2 strain was abnormally long. The CM1 immortalization was 7 times greater than the control and the CM2 strain had the highest frequency of immortalization, 7 times greater than the CM1. These findings indicate that genes associated with chromosome 11p- may be involved in the immortalization of human cells. During their abnormal crisis periods, the cells derived from the patients with Wilms' tumor showed an extremely high frequency of chromosomal aberrations and mutations (6TGs --> 6TGr). These results indicate that when the growth-arrested cells from Wilms' patients are induced to grow with the introduction of SV40LT at the crisis stage they are highly mutable, resulting in their immortalization in vitro.

Antigens, Viral, Tumor↗