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The effect of RAA on mRNA degradation in THP-1 human leukemic monocytes Mak, Isabella Wing Yan
Abstract
Dysregulation of mRNA levels is a genetic hallmark of cancer. The Shaw-Kamen (SK) box, an AU-rich element (ARE ) found in the 3'UTR, is an mRNA stability determinant in many cytokines, growth factors, transcription factors, and proto-oncogene mRNAs. A fungal metabolite, Radicicol Analog A (RAA) , was found to destabilize only S K box-containing mRNAs in THP-1 human leukemic monocytes (Cytokine, 1996, 8, 751-761). Given the small number of genes analyzed in this study, SAGE was used to examine the transcriptome-wide effect of RAA on mRNA expression in THP-1 cells. To determine whether RAA only down-regulated SK box-containing mRNAs, two ~45,000-short-tag SAGE libraries were prepared from total mRNA isolated from THP-1 cells stimulated with IFNγ/LPS ± RAA (lp:M). Surprisingly, only 0.27% of the total unique SAGE tags ( p<0.001) were downregulated by RAA treatment. Tag-to-gene identities for these 48 down-regulated SAGE tags were validated and quantified using real-time RT-PCR. Although 73% of these RAA-down-regulated tags encoded SK box-containing mRNAs, no linear correlation was observed between the fold decrease in mRNA expression and the number of the SK boxes. mRNA half-lives for ten genes, were determined in unstimulated, and 2 and 16 hour post-IFNγ/LPS stimulated THP-1 cells. The mRNA half-lives of two SK box-containing mRNAs, TNF-a and NFKBIA , were unaffected by RAA, suggesting that RAA may affect transcription. Interestingly, all mRNAs stabilized upon IFNγ/LPS treatment were destabilized by RAA at the post-transcriptional level. Hopefully, by studying the effect of RAA on mRNA regulation, a new approach to cancer treatment and other diseases, may be gained.
Item Metadata
Title |
The effect of RAA on mRNA degradation in THP-1 human leukemic monocytes
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Creator | |
Publisher |
University of British Columbia
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Date Issued |
2005
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Description |
Dysregulation of mRNA levels is a genetic hallmark of cancer. The Shaw-Kamen (SK) box,
an AU-rich element (ARE ) found in the 3'UTR, is an mRNA stability determinant in many
cytokines, growth factors, transcription factors, and proto-oncogene mRNAs. A fungal
metabolite, Radicicol Analog A (RAA) , was found to destabilize only S K box-containing
mRNAs in THP-1 human leukemic monocytes (Cytokine, 1996, 8, 751-761). Given the
small number of genes analyzed in this study, SAGE was used to examine the transcriptome-wide
effect of RAA on mRNA expression in THP-1 cells. To determine whether RAA only
down-regulated SK box-containing mRNAs, two ~45,000-short-tag SAGE libraries were
prepared from total mRNA isolated from THP-1 cells stimulated with IFNγ/LPS ± RAA
(lp:M). Surprisingly, only 0.27% of the total unique SAGE tags ( p<0.001) were downregulated
by RAA treatment. Tag-to-gene identities for these 48 down-regulated SAGE tags
were validated and quantified using real-time RT-PCR. Although 73% of these RAA-down-regulated tags encoded SK box-containing mRNAs, no linear correlation was observed
between the fold decrease in mRNA expression and the number of the SK boxes. mRNA
half-lives for ten genes, were determined in unstimulated, and 2 and 16 hour post-IFNγ/LPS
stimulated THP-1 cells. The mRNA half-lives of two SK box-containing mRNAs, TNF-a
and NFKBIA , were unaffected by RAA, suggesting that RAA may affect transcription.
Interestingly, all mRNAs stabilized upon IFNγ/LPS treatment were destabilized by RAA at
the post-transcriptional level. Hopefully, by studying the effect of RAA on mRNA
regulation, a new approach to cancer treatment and other diseases, may be gained.
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Genre | |
Type | |
Language |
eng
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Date Available |
2009-12-10
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Provider |
Vancouver : University of British Columbia Library
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Rights |
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
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DOI |
10.14288/1.0091970
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URI | |
Degree | |
Program | |
Affiliation | |
Degree Grantor |
University of British Columbia
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Graduation Date |
2005-05
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Campus | |
Scholarly Level |
Graduate
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Aggregated Source Repository |
DSpace
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Rights
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.