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Prostate-specific a...
Prostate-specific antigen mRNA and protein levels in laser microdissected cells of human prostate measured by real-time reverse transcriptase-quantitative polymerase chain reaction and immuno-quantitative polymerase chain reaction
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- Pinzani, P. (author)
- Universita degli Studi di Firenze,University of Florence
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- Lind, Kristina, 1977 (author)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Malentacchi, F. (author)
- Universita degli Studi di Firenze,University of Florence
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- Nesi, G. (author)
- Universita degli Studi di Firenze,University of Florence
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- Salvianti, F. (author)
- Universita degli Studi di Firenze,University of Florence
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- Villari, D. (author)
- Universita degli Studi di Firenze,University of Florence
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- Kubista, Mikael, 1961 (author)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Pazzagli, M. (author)
- Universita degli Studi di Firenze,University of Florence
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- Orlando, C. (author)
- Universita degli Studi di Firenze,University of Florence
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(creator_code:org_t)
- Elsevier BV, 2008
- 2008
- English.
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In: Human Pathology. - : Elsevier BV. - 0046-8177. ; 39:10, s. 1474-1482
- Related links:
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http://dx.doi.org/10...
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https://research.cha...
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https://doi.org/10.1...
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Abstract
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- Laser-assisted microdissection has mainly been used in cancer studies to excise pure cell populations from heterogeneous tissues. Cancer and normal cells selected by laser-assisted microdissection have frequently been used for mRNA expression studies usually by reverse transcriptase-quantitative polymerase chain reaction (qPCR). Recently, real time immuno-qPCR was developed as a new tool for highly sensitive measurements of proteins. Using reverse transcriptase-qPCR and immuno-qPCR, we measured the amounts of prostate-specific antigen mRNA and its corresponding protein in homogeneous and comparable cell populations, collected from normal and cancer prostates by laser-assisted microdissection. With these techniques, prostate-specific antigen mRNA and protein were quantified over a wide range of concentrations with a sensitivity sufficient to analyze single prostate cells (LNCaP). We did not find significant differences in prostate-specific antigen protein and mRNA between normal and cancer cells. The expression of prostate-specific antigen protein and mRNA was highly correlated in both normal and pathological cells. In microdissected peritubular stromas areas of prostate cancers, the concentration of prostate-specific antigen protein was about 100 times higher than in normal prostate, indicating an increased transit of secreted prostate-specific antigen. In the same samples, prostate-specific antigen mRNA was not detectable. Our data demonstrate, for the first time, the feasibility of simultaneous application of reverse transcriptase-qPCR and immuno-qPCR in studies of homogeneous cell populations, collected by laser-assisted microdissection. The approach is expected to become a very powerful tool for expression studies in human cancers at both mRNA and protein levels.
Subject headings
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology (hsv//eng)
Publication and Content Type
- art (subject category)
- ref (subject category)
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- By the author/editor
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Pinzani, P.
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Lind, Kristina, ...
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Malentacchi, F.
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Nesi, G.
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Salvianti, F.
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Villari, D.
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show more...
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Kubista, Mikael, ...
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Pazzagli, M.
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Orlando, C.
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show less...
- About the subject
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- ENGINEERING AND TECHNOLOGY
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ENGINEERING AND ...
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and Industrial Biote ...
- Articles in the publication
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Human Pathology
- By the university
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Chalmers University of Technology