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Analysis of triptolide-regulated gene expression in Jurkat cells by complementary DNA microarray
Ze-Ying Du1, Xiao-Yu Li, Yuan-Chao Li
1Department of Pharmacology, Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Aim:
To investigate the global gene expression profile changes in Jurkat cells after triptolide treatment in order to find the possible triptolide targets.
Methods:
Jurkat cells were treated with or without triptolide 10 microg/L for 2 h. Total RNA were isolated and used as templates for reverse transcriptional labeling of fluorescent cDNA probes. High density DNA microarray chips with a set of 13,872 human genes/Ests were used to generate the expression profile of triptolide-treated or untreated control Jurkat cells by hybridizing with fluorescent labeled probes. Array image was acquired and analyzed with array analyzing software GeneSpring.
Results:
Triptolide significantly suppressed expression of 117 genes in Jurkat cells. Among these 117 genes, 30 % were Ests or genes without known functions, 13 % were transcription factors, 9 % were signal transduction pathway regulators, and 9 % were DNA binding proteins. Notably, the expression of mitogen-activated protein kinase kinase kinase kinase 5 (MAP kinase 5) and phosphoinositide-3-kinase (PI-3 kinase) was inhibited more than 100-fold. Moreover, the expression of genes involved in lipid transportation and metabolism was down-regulated by triptolide.
Conclusion:
High-density microarray provided an effective approach to identify drug targeting molecules. It is suggested that the widely known immune suppressive and antitumor effects of triptolide were mediated at least in part by suppression of MAP kinase and PI-3 kinase gene expression.
Insights
Triptolide treatment significantly altered gene expression in Jurkat cells, suppressing 117 genes, including key signaling pathways like MAP kinase and PI-3 kinase. This reveals potential drug targets for triptolide
Area of Science:
- Molecular Biology
- Genomics
- Pharmacology
Background:
- Triptolide is known for its immunosuppressive and antitumor properties.
- Identifying the molecular targets of triptolide is crucial for understanding its mechanisms of action.
Purpose of the Study:
- To investigate global gene expression changes in Jurkat cells following triptolide treatment.
- To identify potential molecular targets of triptolide through gene expression profiling.
Main Methods:
- Jurkat cells were treated with triptolide (10 microg/L) for 2 hours.
- Total RNA was isolated and used for cDNA probe synthesis.
- High-density DNA microarrays (13,872 genes/Ests) were employed to analyze gene expression profiles.
- GeneSpring software was used for array image analysis.
Main Results:
- Triptolide significantly suppressed the expression of 117 genes in Jurkat cells.
- Downregulated genes included transcription factors, signal transduction regulators, and DNA-binding proteins.
- Notably, mitogen-activated protein kinase kinase kinase kinase 5 (MAP kinase 5) and phosphoinositide-3-kinase (PI-3 kinase) expression decreased over 100-fold.
- Genes involved in lipid transport and metabolism were also downregulated.
Conclusions:
- High-density microarrays are effective for identifying drug targets.
- Triptolide's immunosuppressive and antitumor effects may be mediated by the suppression of MAP kinase and PI-3 kinase gene expression.
