ABSTRACT
DNA chip has been used as a powerful tool to study the genetic reprogramming of cells and its link to cellular phenotype such as angiogenesis. To evaluate the angiogenesis related genetic reprogramming more efficiently, we here developed an angiogenesis- focused cDNA chip containing 153 angiogenesis related genes arrayed in duplicate on a slide glass. In order to validate the functionality of the angiogenesis-focused cDNA chip, we examined gene expression profiles in HT1080 cells treated with either fetal bovine serum, a well known pro-angiogenic factor, or trichostatin A, a known angiogenesis inhibitor, using the cDNA chip. All duplicate data from the analysis are well matched with each other and gene expression profiles are well consistent with previously reported data. These results demonstrate that the angiogenesis-focused cDNA chip developed here can be a useful tool towards angiogenesisrelated researches.
Subject(s)
Humans , Angiogenesis Inducing Agents/pharmacology , Angiogenesis Inhibitors/pharmacology , Gene Expression/drug effects , Gene Expression Profiling/instrumentation , Neovascularization, Pathologic/genetics , Neovascularization, Physiologic/genetics , Oligonucleotide Array Sequence Analysis/instrumentation , Tumor Cells, CulturedABSTRACT
Calmegin is a testis-specific molecular chaperon playing a key role in spermatogenesis. However, the transcriptional regulatory mechanisms for calmegin expression are entirely unknown. Herein, we revealed that calmegin is transcriptionally regulated by histone deacetylase (HDAC) and CpG methyltransferase. The cDNA microarray analysis of the human fibrosarcoma cells treated with trichostatin A (TSA) showed an increased level of calmegin mRNA. The induction of calmegin mRNA by TSA was added by the treatment with 5-aza-2'-deoxycytidine (5'Aza- dC), implying that epigenetic alterations are involved in the transcriptional repression of the gene. Moreover, chromatin immunoprecipitation assay using an anti-acetyl-histone H3 antibody exhibited that the proximal region (-152~-31) of the calmegin promoter is responsible for HDAC-mediated transcriptional repression of the gene. These results demonstrate that calmegin expression is regulated by HDAC and CpG methyltransferase in a coordinative way.
Subject(s)
Animals , Humans , Male , Mice , Calcium-Binding Proteins/genetics , Cell Line, Tumor , Gene Expression Regulation , Histone Deacetylases/metabolism , Methyltransferases/metabolism , Molecular Chaperones/genetics , Organ Specificity , Promoter Regions, Genetic/genetics , Testis/metabolism , Transcription, GeneticABSTRACT
In the course of screening of angiogenesis inhibitor from natural products, cryptotanshinone from Salvia miltiorrhiza was isolated as a potent small molecule inhibitor of angiogenesis. Cryptotanshinone inhibits bFGF-induced angiogenesis of BAECs at ten micromolar ranges in vitro without cytotoxicity. Tanshinone IIA, another tanshinone isolated from S. miltiorrhiza, which is structurally very similar to cryptotanshinone except C-15 position of dihydrofuran ring does not inhibit angiogenesis induced by bFGF. These results demonstrate that cryptotanshinone is a new anti-angiogenic agent and double bond at C-15 position of the dihydrofuran ring plays a crucial role in the activity.
Subject(s)
Animals , Cattle , Humans , Angiogenesis Inhibitors/chemistry , Cell Movement/drug effects , Cell Proliferation/drug effects , Cells, Cultured , Drugs, Chinese Herbal/chemistry , Endothelial Cells/drug effects , Fibroblast Growth Factor 2/pharmacology , Neovascularization, Physiologic/drug effects , Phenanthrenes/chemistry , Plant Roots/chemistry , Salvia miltiorrhiza/chemistryABSTRACT
The methanolic extract of fruiting body of Paecilomyces tenuipes DGUM 32001 showed significant cytotoxicity against human cancer cells: HepG2 and MCF-7. The methanolic extract was further fractionated with organic solvents such as chloroform and ethyl acetate in that order. Among the fractions tested, the ethyl acetate fraction showed the highest cytotoxicity against the carcinoma tested. The IC50 values of ethyl acetate fraction against HepG and MCF-7 were 40 and 9.6 microg/ml, respectively.