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Comparative Time-Dependent Analysis of Potential Inflammation Biomarkers in Lymphoma-Bearing SJL Mice.

Kåredal, Monica LU orcid ; Bhat, Vadiraja ; Babu, I ; Wishnok, John and Tannenbaum, Steven (2007) In Journal of Proteome Research 6(5). p.1735-1744
Abstract
SJL mice colonized with RcsX lymphoma cells undergo a rapid inflammatory response associated with biological and physiological effects including increased nitric oxide production and mutations in spleen DNA. By 2 weeks postcolonization, these changes were accompanied by both up- and down-regulation of a number of plasma proteins. In the experiments reported here, plasma from individual SJL mice was analyzed at several time-points over the 2-week period to determine if there were sets of proteins whose expression varied in concert and thus might serve as early biomarkers for inflammation-related disorders. Samples were collected just prior to injection of the RcsX cells and then after 4, 8, and 12 days. Albumin and immunoglobulins were... (More)
SJL mice colonized with RcsX lymphoma cells undergo a rapid inflammatory response associated with biological and physiological effects including increased nitric oxide production and mutations in spleen DNA. By 2 weeks postcolonization, these changes were accompanied by both up- and down-regulation of a number of plasma proteins. In the experiments reported here, plasma from individual SJL mice was analyzed at several time-points over the 2-week period to determine if there were sets of proteins whose expression varied in concert and thus might serve as early biomarkers for inflammation-related disorders. Samples were collected just prior to injection of the RcsX cells and then after 4, 8, and 12 days. Albumin and immunoglobulins were depleted, and the samples were resolved by 1D gel electrophoresis. The gels were cut into 20 slices, and the proteins were digested in-gel with trypsin. The digests were treated with iTRAQ reagents and then analyzed using LC/MS/MS. The resulting data were processed with two software packages, that is, ProQuant and Spectrum Mill, and then subjected to K-means cluster analysis (K = 4). The four clusters revealed a set of highly up-regulated proteins, a set of progressively up-regulated proteins, a set with no major changes, and a set that declined. The first cluster included haptoglobin and serum amyloid A; the second included groups with several functions including protease inhibition, cell motility, and transport. The iTRAQ results for a selection of the up-regulated proteins, including haptoglobin, hemopexin, serum amyloid P component, and ceruloplasmin, were confirmed with Western blots. Prominent down-regulated proteins included esterase-1, paraoxonase, and alpha-2-macroglobulin. Approximately 50% of the up-regulated proteins are canonical acute phase proteins, while the remainder are regulated by the Nrf2 transcription factor. Keywords: inflammation center dot SJL mouse center dot lymphoma center dot iTRAQ center dot tumor progression center dot biomarker center dot cluster analysis center dot acute phase proteins (Less)
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author
; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
biomarker, tumor progression, iTRAQ, SJL mouse, inflammation, lymphoma, cluster analysis, acute phase proteins
in
Journal of Proteome Research
volume
6
issue
5
pages
1735 - 1744
publisher
The American Chemical Society (ACS)
external identifiers
  • wos:000246207000010
  • scopus:34249314688
ISSN
1535-3893
DOI
10.1021/pr060497x
language
English
LU publication?
yes
id
dbde710f-a262-49a9-b4f2-3883a1dc8530 (old id 166196)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=17388619&dopt=Abstract
date added to LUP
2016-04-01 11:42:48
date last changed
2022-02-10 20:25:16
@article{dbde710f-a262-49a9-b4f2-3883a1dc8530,
  abstract     = {{SJL mice colonized with RcsX lymphoma cells undergo a rapid inflammatory response associated with biological and physiological effects including increased nitric oxide production and mutations in spleen DNA. By 2 weeks postcolonization, these changes were accompanied by both up- and down-regulation of a number of plasma proteins. In the experiments reported here, plasma from individual SJL mice was analyzed at several time-points over the 2-week period to determine if there were sets of proteins whose expression varied in concert and thus might serve as early biomarkers for inflammation-related disorders. Samples were collected just prior to injection of the RcsX cells and then after 4, 8, and 12 days. Albumin and immunoglobulins were depleted, and the samples were resolved by 1D gel electrophoresis. The gels were cut into 20 slices, and the proteins were digested in-gel with trypsin. The digests were treated with iTRAQ reagents and then analyzed using LC/MS/MS. The resulting data were processed with two software packages, that is, ProQuant and Spectrum Mill, and then subjected to K-means cluster analysis (K = 4). The four clusters revealed a set of highly up-regulated proteins, a set of progressively up-regulated proteins, a set with no major changes, and a set that declined. The first cluster included haptoglobin and serum amyloid A; the second included groups with several functions including protease inhibition, cell motility, and transport. The iTRAQ results for a selection of the up-regulated proteins, including haptoglobin, hemopexin, serum amyloid P component, and ceruloplasmin, were confirmed with Western blots. Prominent down-regulated proteins included esterase-1, paraoxonase, and alpha-2-macroglobulin. Approximately 50% of the up-regulated proteins are canonical acute phase proteins, while the remainder are regulated by the Nrf2 transcription factor. Keywords: inflammation center dot SJL mouse center dot lymphoma center dot iTRAQ center dot tumor progression center dot biomarker center dot cluster analysis center dot acute phase proteins}},
  author       = {{Kåredal, Monica and Bhat, Vadiraja and Babu, I and Wishnok, John and Tannenbaum, Steven}},
  issn         = {{1535-3893}},
  keywords     = {{biomarker; tumor progression; iTRAQ; SJL mouse; inflammation; lymphoma; cluster analysis; acute phase proteins}},
  language     = {{eng}},
  number       = {{5}},
  pages        = {{1735--1744}},
  publisher    = {{The American Chemical Society (ACS)}},
  series       = {{Journal of Proteome Research}},
  title        = {{Comparative Time-Dependent Analysis of Potential Inflammation Biomarkers in Lymphoma-Bearing SJL Mice.}},
  url          = {{http://dx.doi.org/10.1021/pr060497x}},
  doi          = {{10.1021/pr060497x}},
  volume       = {{6}},
  year         = {{2007}},
}