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Induction of apoptosis by Solenopsin B: evalutaion by DNA microarray and RT 2 Profiler™ PCR array analysis.
Author(s) -
Sample Robert Cressten,
Sullivan Donna C,
Kelley Kashonda,
Rockhold Robin
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.526.15
Subject(s) - dna laddering , apoptosis , microbiology and biotechnology , biology , cell cycle , dna microarray , microarray analysis techniques , microarray , programmed cell death , complementary dna , gene , cell , gene expression , dna fragmentation , genetics
The effect of synthetic fire ant venom alkaloid Solenopsin B (Sol B) on the human monocytic cell line U937 was examined to determine its ability to induce apoptosis and efficacy as a therapeutic agent. Sol B treated cells displayed a >50% reduction in viability along with DNA laddering, a hallmark of apoptosis. The apoptosis mechanism was further examined using DNA microarrays and quantitative RT‐PCR (qRT‐PCR). U937 cells were incubated in the presence of Sol B and total cellular RNA isolated. cDNA was synthesized, labeled with cy3/cy5 and hybridized to microarrays or used in qRT‐PCR RT 2 Profiler(tm) PCR array analysis. Microarray analysis revealed that 661 genes and 620 ESTs were up regulated >1.5 fold, including several apoptosis and cell cycle genes. PCR arrays representing functional gene groupings of 90 apoptosis and 92 cell cycle genes showed up regulation of >90% and >70% respectively. Genes examined using both microarray and qRT‐PCR showed correlations of ~93% for apoptosis and ~79% for cell cycle genes. Further, transmission electron microscopy revealed Sol B treatment resulted in loss of cell membrane integrity further verifying the ability of Sol B to induce apoptosis. Collectively these findings indicate that Sol B induced programmed cell death in human cells by triggering the apoptotic pathway.