Investigation into mechanisms mediating the inhibitory effect of 1,4-benzodiazepines on mast cells by gene expression profiling.
OBJECTIVE
This study aims to identify by a molecular genetic approach potential targets in mast cells at which 1,4-benzodiazepines may cause their inhibitory effect on mast cell activity.


METHODS
Gene expression analyses with microarray gene chip and/or quantitative PCR were performed using 1,4-benzodiazepine-treated human mast cell leukemia HMC-1.2 cells, promyelocytic leukemia HL-60 cells and human mast cells from healthy volunteers and patients with mast cell activation disease (MCAD). Pathway analysis was applied to search for enriched biological functions and canonical pathways within differentially regulated genes.


RESULTS
Both neoplastic and normal human mast cells express several GABA(A) receptor subunits at the mRNA level. In mast cells from MCAD patients expression of some GABA(A) receptor subunits and expression of the translocator protein TSPO are increased compared with those from healthy controls. Expression of the protein tyrosine kinases Lyn, Fgr and Yes1 was increased in HMC-1.2 cells as compared with the ontogenetically related HL60 cells. Differences in gene regulation in HMC-1.2 cells after treatment with the 1,4-benzodiazepines clonazepam, flunitrazepam and 4-chlorodiazepam suggested that signaling and gene expression induced by clonazepam was similar to that of flunitrazepam but different from that of 4-chlorodiazepam. This conclusion is supported by the results of the pathway analysis.


CONCLUSIONS
A novel type of GABA(A) receptors on mast cells appears to be involved in the inhibition of mast cell activity by 1,4-benzodiazepines. These receptors seem to be composed without γ subunits suggesting unique pharmacological properties. An action at Src-kinases, or at TSPO located in the plasma membrane may also be involved.
Institute of Human Genetics, University of Bonn, Bonn, Germany.
Britta
B
Haenisch
Michael
M
Huber
Thomas
T
Wilhelm
Michael
M
Steffens
Gerhard J
GJ
Molderings
D000328:Adult; D000368:Aged; D001570:Benzodiazepinones / Q000494:pharmacology*; D002998:Clonazepam / Q000494:pharmacology*; D005260:Female; D005445:Flunitrazepam / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling / Q000379:methods; D018922:HL-60 Cells; D006801:Humans; D008297:Male; D008407:Mast Cells / Q000187:drug effects*; D008415:Mastocytosis / Q000235:genetics; D046228:Microarray Analysis / Q000379:methods; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods
D001570:Benzodiazepinones; C028513:4'-chlorodiazepam; D002998:Clonazepam; D005445:Flunitrazepam