[Transfection of TF-1 cell line with human bcl-2 proto-oncogene provides short-term survival in absence of GM-CSF without changing the phenotype].
Apoptosis is a process genetically controlled. The produce of the bcl-2 gene, bcl-2, is an anti apoptotic protein that is linked to the external membrane of the mitochondria.


OBJECTIVE
To explore the possibility that bcl-2 transfection could change phenotype, response to mitogenic factor, and cell morphology on the TF-1 parental cell line and the bcl-2 transfectant TB-1 or TF-1neo.


METHODS
We look at the expression of CD13, CD34 and c-Kit surface markers by flow cytometry. We have measured cell proliferation in response to GM-CSF and cell survival after GM-CSF withdrawal by the MTT assay on the same cell lines. Apoptosis was evaluated by the apoptotic membrane blebbing set up at different times after serum and survival factor removal or tolerance to cytotoxic compounds from Justicia spicigera.


RESULTS
According with our results, ectopic expression of the bcl-2 gene prevented apoptosis without changes in morphology or phenotype in the absence of GM-CSF and serum or the presence of the extract from Justicia spicigera. Consisting with the Bcl-2 function, we found that Bcl-2 did not change response to GM-CSF. Serum deprivation or GM-CSF withdrawal induces cell death at 36 hours in TF-1 and TF-1neo cells, whereas TB-1 cells undergo apoptotic membrane blebbing after 96 hours under the same conditions.


CONCLUSIONS
Taken together, our data indicate that Bcl-2 is a short term anti apoptotic protein in TB-1 cell line, that does not affect response to GM-CSF neither CD13, CD34 nor c-Kit antigen expression.
Alvarado-Moreno
J A
JA
Facultad de Estudios Superiores Zaragoza, Universidad Nacional Autónoma de México.
Nieves-Ramírez
M E
ME
Cáceres-Cortés
J R
JR
D002460:Cell Line; D002470:Cell Survival; D019254:Genes, bcl-2 / Q000235:genetics*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006801:Humans; D010641:Phenotype; D000090063:Proto-Oncogene Mas; D013997:Time Factors; D014162:Transfection
C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D016178:Granulocyte-Macrophage Colony-Stimulating Factor