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| Status |
Public on Feb 28, 2009 |
| Title |
Sanguinarine, putative anticancer therapeutic, interacts,modulates and transcripts in the context of chromatin |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
DNA binding anticancer agents cause alteration in chromatin structure and dynamics. Here, we report the dynamic interaction of the DNA intercalator and potential anticancer, plant alkaloid, Sanguinarine (SGR) with chromatin. Association of SGR with different levels of chromatin structure was found to be enthalpydriven. Apart from DNA it binds with comparable affinity to histones and induces chromatin aggregation. The dual binding property of SGR leads to inhibition of core histone modifications. Although, it potently inhibits H3K9 methylation by G9a in vitro, H3K4 and H3R17 methylation are more profoundly inhibited in cells. SGR inhibits histone acetylation both in vitro and in vivo. It does not affect the in vitro transcription from DNA template but significantly represses acetylation dependent chromatin transcription. SGR mediated repression of epigenetic marks and the alteration of chromatin geography, also result in modulation of global gene expression. These data conclusively, show the anticancer DNA binding intercalator as a modulator of chromatin modifications and transcription in the chromatin context.
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| Overall design |
The total RNA was isolated from control and treated cells using TRIZOL (Invitrogen) method. The Micromax direct labeling kit, MPS502 (PerkinElmer) was used to synthesize the labeled cDNA from 70 ug of total RNA and further process the hybridized cDNA on the array. All the steps were carried out according to manufacturer’s instructions (www.perkinelmer.com/lifesciences). The array slides were scanned immediately by PerkinElmer Scan array Gx Microarray scanner. The Scan array software (PerkinElmer) was used for grid wise normalization of array images. Two arrays were used with at least one biological treatment of cells and dye swap experiment were included in the final analysis. . The data was analysed by GeneSpring GX and Biointerpreter software from Genotypic Technology, Bangalore. The differential expression was considered if the Log 2 mean of at least -1 for the down regulated genes and +1 for the upregulated genes. We considered only the genes that were reproducible from both replicates.
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| Contributor(s) |
Kundu T, BR S |
| Citation(s) |
19246011 |
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| Submission date |
Jan 07, 2009 |
| Last update date |
Mar 20, 2012 |
| Contact name |
Tapas Kumar Kundu |
| E-mail(s) |
tapas@jncasr.ac.in
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| Phone |
+91-80-22082840
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| Fax |
+91-80-22082866
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| URL |
http://www.jncasr.ac.in/tdl/
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| Organization name |
Jawaharlal Nehru Centre for Advanced scientific Research
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| Department |
Molecular Biology and Genetics Unit
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| Lab |
Transcription and Disease Laboratory
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| Street address |
Jakkur
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| City |
Bangalore |
| State/province |
Karnataka |
| ZIP/Postal code |
560064 |
| Country |
India |
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| Platforms (1) |
| GPL8032 |
Human Agilent-016332 custom array |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA111315 |
