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Proteomic analysis of laser capture ...
~
Paweletz, Cloud Peter.
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Proteomic analysis of laser capture microdissected esophageal and prostatic cells during tumorigenesis.
紀錄類型:
書目-語言資料,印刷品 : Monograph/item
正題名/作者:
Proteomic analysis of laser capture microdissected esophageal and prostatic cells during tumorigenesis./
作者:
Paweletz, Cloud Peter.
面頁冊數:
270 p.
附註:
Mentors: Paul Roepe; Emanuel F. Petricoin; Lance A. Liotta.
Contained By:
Dissertation Abstracts International63-05B.
標題:
Chemistry, Biochemistry. -
電子資源:
http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3053329
ISBN:
0493677747
Proteomic analysis of laser capture microdissected esophageal and prostatic cells during tumorigenesis.
Paweletz, Cloud Peter.
Proteomic analysis of laser capture microdissected esophageal and prostatic cells during tumorigenesis.
- 270 p.
Mentors: Paul Roepe; Emanuel F. Petricoin; Lance A. Liotta.
Thesis (Ph.D.)--Georgetown University, 2002.
Laser Capture Microdissection (LCM) allows for the procurement of pure cell populations under direct microscopic visualization. In this regard important clinical and molecular benefits are realized if the protein content between normal and diseased cell populations procured from human tissue are analyzed. Not only does one obtain pure and isogeneic cell populations with no contaminating influences, but more importantly, direct intra-patient comparisons of protein expression fluxes are assessed. With such pure cell populations available low-, medium-, and high-throughput assays were developed that allow for discovery and validation of protein targets and protein patterns for potential therapeutic or diagnostic purposes.
ISBN: 0493677747Subjects--Topical Terms:
1017722
Chemistry, Biochemistry.
Proteomic analysis of laser capture microdissected esophageal and prostatic cells during tumorigenesis.
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Mentors: Paul Roepe; Emanuel F. Petricoin; Lance A. Liotta.
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Laser Capture Microdissection (LCM) allows for the procurement of pure cell populations under direct microscopic visualization. In this regard important clinical and molecular benefits are realized if the protein content between normal and diseased cell populations procured from human tissue are analyzed. Not only does one obtain pure and isogeneic cell populations with no contaminating influences, but more importantly, direct intra-patient comparisons of protein expression fluxes are assessed. With such pure cell populations available low-, medium-, and high-throughput assays were developed that allow for discovery and validation of protein targets and protein patterns for potential therapeutic or diagnostic purposes.
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First, global protein expression of LCM procured cell populations were assessed by two-dimensional electrophoresis followed by subsequent identification by tandem mass spectrometry. 50000 microdissected cells resolved 675 distinct proteins with molecular weights ranging from 18 kDa to 200 kDa. One of these proteins consistently differently expressed via 2D-PAGE analysis in esophageal and prostate cancer was identified as annexin-I. Further analysis showed that annexin-I protein expression decreased with loss of differentiation in esophageal cell populations.
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However 2D-PAGE is inherently low throughput, time and labor intensive. Therefore I developed protein microarrays that allow for screening of molecular markers and pathway targets in a more effective and higher throughput assay relative to 2D-PAGE. Specifically, our protein arrays were designed in such a way that the activation status of a specific candidate molecule is readily assessed. Using this protein microarray prostate cancer progression was associated with increased phosphorylation of Akt, suppression of apoptosis pathways, and decreased activation of ERK.
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Lastly, mass spectrometric chromatograms from microdissected cell lysates and serum were generated by high-throughput surface enhanced laser desorption and ionization spectrometry. Consistent protein changes were identified in microdissected longitudinally matched esophageal and prostate cell populations. Heuristical analysis of chromatograms generated from serum samples between healthy and individuals with biopsy proven cancer of the prostate yielded protein patterns that correlate with early diagnosis.
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http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3053329
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