Inducible functional expression of Bcl-2 in human astrocytes derived from NTera-2 cells.
Astrocytes provide structural support for neurons and may also play important roles in both neuroprotection and neurodegeneration. We, here report that human astrocytes derived from on NTera-2 (NT2) cell line expressing a functional anti-apoptotic protein bcl-2 under the control of a tetracycline responsive promoter using the Tet-On and Tet-Off expression systems. NT2 cells were transfected with the Tet On or Tet Off vectors followed by pTRE carrying human bcl-2. Drug resistant cells were differentiated into astrocytes under retinoic acid exposure. These astrocyte lines were found to express astrocyte specific markers such glial fibrillary acidic protein and chemokine receptors (CCR5, CXCR4) but not CCR3 and CD4. Furthermore, NT2 astrocytes expressing bcl-2 showed rapid response to doxycycline presence in the Tet On and Tet off system. The inducible expression of bcl-2 was found to be tightly regulated by doxycycline concentration in the NT2 astrocytes. We also showed that the induction of bcl-2 expression prevented NT2 astrocytes from camptothecin-induced cellular damage. These results indicate that this system may be useful for the study of specific effects of bcl-2 gene expression on astrocyte function(s) and cellular damage.
Ozdener
Hakan
H
University of Pennsylvania, 3400 Spruce Street, Philadelphia, PA 19104, USA. hozdener@yahoo.com
D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D001253:Astrocytes / Q000378:metabolism*; D015153:Blotting, Western; D002166:Camptothecin / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D004247:DNA / Q000235:genetics; D053938:DNA Fragmentation; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000235:genetics / Q000502:physiology; D019254:Genes, bcl-2 / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D011401:Promoter Regions, Genetic / Q000187:drug effects; D019707:Receptors, Chemokine / Q000187:drug effects; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013752:Tetracycline / Q000494:pharmacology; D014162:Transfection
D000972:Antineoplastic Agents, Phytogenic; D019707:Receptors, Chemokine; D004247:DNA; D013752:Tetracycline; D002166:Camptothecin