J. Biochem, 1993, Vol. 114, No. 2 194-202
© 1993 Japanese Biochemical Society
other |
Isolation of a Subclonal Cell Line of PC12 Transfected with Dexamethasone-Regulated ras Oncogene: Morphological Differentiation, Biochemical Properties, and Tumorigenicity1
*Department of Biochemistry, School of Medicine, Kitasato University Sagamihara, Kanagawa 228
**Department of Microbiology, School of Hygienic Sciences, Kitasato University Sagamihara, Kanagawa 228
***Department of Functional Morphology, School of Nursing, Kitasato University Sagamihara, Kanagawa 228
****Department of Medical Informatics, School of Medicine, Kitasato University Sagamihara, Kanagawa 228
*****S.M.I. Bristol Co. Sagamihara, Kanagawa 228
We have isolated and characterized a new subclonal cell line designated as MR31, which was obtained by transfection of PC12 cells with a glucocorticoid-regulated ras oncogene. The mRNA derived from the c-Ha-ras gene was proved to be expressed on exposure of the MR31 cells to dexamethasone, the highest value being attained at 8 h. MR31 cells rapidly extended neurite-like processes within 24 h in response to dexamethasone as well as nerve growth factor (NGF). The time of onset of neurite outgrowth induced by dexamethasone corresponded to the time when the highest ras mRNA level was observed. The cate-cholamine content of MR31 cells was found to be twice that of PC12 cells. A time course study on the effects of dexamethasone or NGF on cells showed that the former caused an increase in dopamine, a major catecholamine, to twofold the control level at 48 h after the treatment, while the latter caused a decrease in the dopamine level. These effects on catecholamines were almost the same in MR31 and PC12 cells. The acetylcholinesterase activity of MR31 cells was enhanced by both dexamethasone and NGF, whereas that of PC12 cells was enhanced by NGF, but not by dexamethasone. The changes in acetylcholinesterase activity were correlated with neurite outgrowth. Electron-microscopically, MR31 cells were not different from PC12 cells. MR31 cells exhibited extremely decreased tumorigenicity as compared with PC12 cells. The morphological and biochemical properties of MR31 cells remained constant, even after repeated passages. These findings indicate that MR31 cells represent a stable transfectant exhibiting different properties from those of the parent PC12 cells and are at the early stage of differentiating PC12 cells, characterized by rapid response to NGF and dexamethasone.
1This study was supported in part by a Grant-in-Aid for Scientific Research (No. 62580130) and Grant-in-Aid for Scientific Research on Priority Areas (No. 02259212) from the Ministry of Education, Science and Culture of Japan.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  M. Fukuzumi, S. Maruyama, M. Sano, and S. Fukui Comparison of the expression of cell surface poly-N-acetyllactosamine-type oligosaccharides in PC12 cells with those in its variant PC12D Glycobiology, June 1, 2001; 11(6): 481 - 494. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Gendron*, L. Laflamme*, N. Rivard, C. Asselin, M. D. Payet, and N. Gallo-Payet Signals from the AT2 (Angiotensin Type 2) Receptor of Angiotensin II Inhibit p21ras and Activate MAPK (Mitogen-Activated Protein Kinase) to Induce Morphological Neuronal Differentiation in NG108-15 Cells Mol. Endocrinol., September 1, 1999; 13(9): 1615 - 1626. [Abstract] [Full Text]
|
|