Identification of genes involved in Ca2+ ionophore A23187-mediated apoptosis and demonstration of a high susceptibility for transcriptional repression of cell cycle genes in B lymphoblasts from a patient with Scott syndrome.
BACKGROUND
In contrast to other agents able to induce apoptosis of cultured cells, Ca2+ ionophore A23187 was shown to elicit direct activation of intracellular signal(s). The phenotype of the cells derived from patients having the hemorrhagic disease Scott syndrome, is associated with an abnormally high proportion of apoptotic cells, both in basal culture medium and upon addition of low ionophore concentrations in long-term cultures. These features are presumably related to the mutation also responsible for the defective procoagulant plasma membrane remodeling. We analyzed the specific transcriptional re-programming induced by A23187 to get insights into the effect of this agent on gene expression and a defective gene regulation in Scott cells.


RESULTS
The changes in gene expression upon 48 hours treatment with 200 nM A23187 were measured in Scott B lymphoblasts compared to B lymphoblasts derived from the patient's daughter or unrelated individuals using Affymetrix microarrays. In a similar manner in all of the B cell lines, results showed up-regulation of 55 genes, out of 12,000 represented sequences, involved in various pathways of the cell metabolism. In contrast, a group of 54 down-regulated genes, coding for histones and proteins involved in the cell cycle progression, was more significantly repressed in Scott B lymphoblasts than in the other cell lines. These data correlated with the alterations of the cell cycle phases in treated cells and suggested that the potent effect of A23187 in Scott B lymphoblasts may be the consequence of the underlying molecular defect.


CONCLUSIONS
The data illustrate that the ionophore A23187 exerts its pro-apoptotic effect by promoting a complex pattern of genetic changes. These results also suggest that a subset of genes participating in various steps of the cell cycle progress can be transcriptionally regulated in a coordinated fashion. Furthermore, this research brings a new insight into the defect in cultured Scott B lymphoblasts, leading to hypothesize that a mutated gene plays a role not only in membrane remodeling but also in signal transduction pathway(s) leading to altered transcriptional regulation of cell cycle genes.
Kozian
Detlef
D
Therapeutic Department Thrombosis and Angiogenesis, Aventis Pharma Germany (Sanofi-Aventis Group), Industriepark Hoechst, Building H831, 65926 Frankfurt, Germany. Detlef.Kozian@sanofi-aventis.com
Proulle
Valérie
V
Nitsche
Almut
A
Galitzine
Marie
M
Martinez
Marie-Carmen
MC
Schumann
Beatrice
B
Meyer
Dominique
D
Herrmann
Matthias
M
Freyssinet
Jean-Marie
JM
Kerbiriou-Nabias
Danièle
D
D017209:Apoptosis; D001402:B-Lymphocytes / Q000378:metabolism*; D000001:Calcimycin / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D002453:Cell Cycle; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D016000:Cluster Analysis; D003029:Coagulants / Q000378:metabolism; D015536:Down-Regulation; D005786:Gene Expression Regulation; D006474:Hemorrhagic Disorders / Q000097:blood*; D006657:Histones / Q000378:metabolism; D006801:Humans; D007476:Ionophores / Q000494:pharmacology*; D046228:Microarray Analysis; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D018075:RNA, Complementary / Q000378:metabolism; D015398:Signal Transduction; D013577:Syndrome; D014158:Transcription, Genetic; D015854:Up-Regulation
D003029:Coagulants; D006657:Histones; D007476:Ionophores; D018075:RNA, Complementary; D000001:Calcimycin; D002118:Calcium