Effects of altered cyclophilin A expression on growth and differentiation of human and mouse neuronal cells.
1. Cyclophilin A (CyP-A), a soluble cytoplasmic immunophilin, is known for its involvement in T cell differentiation and proliferation. Although CyP-A has a pivotal role in the immune response, it is most highly concentrated in brain, where its functions are largely unknown. 2. We reported previously that a murine neuroblastoma (NB-P2) cell line can partially differentiate into neurons when treated with cyclosporin A (CyS-A), implicating a role for CyP-A in neuronal differentiation (Hovland et al. [1999]. Neurochem. Int. 3:229-235). 3. The role of CyP-A in regulating neuronal growth and differentiation is not well defined. To investigate this, we first tested the utility of retroviral-mediated gene transfer and expression in human embryonic brain (HEB) and NB-P2 cells. Second, we examined the effects of retroviral-mediated overexpression or antisense-mediated reduction of CyP-A in HEB and NB-P2 cells. 4. Our data show that retroviral vectors are efficient for stable gene transfer and expression in both cell lines. Moreover, neither overexpression nor reduction of CyP-A expression in NB-P2 cells altered the growth rate or induced differentiation. More importantly, the up-or down-regulation of CyP-A expression did not affect the magnitude of cAMP-induced NB-P2 differentiation. However, overexpression of CyP-A increased the growth rate of HEB cells. 5. In summary, the utility of retroviral vectors for stable gene expression in human embryonic brain and murine neuroblastoma cells was shown. Furthermore, a novel role for CyP-A in augmenting the proliferation of human embryonic brain cells was demonstrated in vitro.
Nahreini
P
P
Center for Vitamins and Cancer Research, Department of Radiology, School of Medicine, University of Colorado Health Sciences Center, Denver 80262, USA. piruz.nahreini@uchsc.edu
Hovland
A R
AR
Kumar
B
B
Andreatta
C
C
Edwards-Prasad
J
J
Prasad
K N
KN
D000818:Animals; D001921:Brain / Q000166:cytology; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D000242:Cyclic AMP / Q000502:physiology; D021984:Cyclophilin A / Q000235:genetics*; D016373:DNA, Antisense; D005333:Fetus / Q000166:cytology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D009447:Neuroblastoma; D009457:Neuroglia / Q000166:cytology / Q000502:physiology; D009474:Neurons / Q000166:cytology* / Q000201:enzymology*; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured
D016373:DNA, Antisense; D000242:Cyclic AMP; D021984:Cyclophilin A