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

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 pol...

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Veröffentlicht in:	Human pathology Jg. 39; H. 10; S. 1474 - 1482
Hauptverfasser:	Pinzani, Pamela, Lind, Kristina, Malentacchi, Francesca, Nesi, Gabriella, Salvianti, Francesca, Villari, Donata, Kubista, Mikael, Pazzagli, Mario, Orlando, Claudio
Format:	Journal Article
Sprache:	Englisch
Veröffentlicht:	New York, NY Elsevier Inc 01.10.2008 Elsevier Elsevier Limited
Schlagworte:	Acids Adenocarcinoma - genetics Adenocarcinoma - metabolism Adenocarcinoma - pathology Antibodies, Monoclonal - immunology Biological and medical sciences Cell Line, Tumor Enzyme-Linked Immunosorbent Assay Gene expression Gene Expression Regulation, Neoplastic Humans Hypotheses Immuno-qPCR Investigative techniques, diagnostic techniques (general aspects) Laser microdissection Lasers Male Medical sciences Methods Microdissection Pathology Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques Polymerase chain reaction Polymerase Chain Reaction - methods Prostate - metabolism Prostate - pathology Prostate cancer Prostate specific antigen Prostate-Specific Antigen - genetics Prostate-Specific Antigen - metabolism Prostatic Neoplasms - genetics Prostatic Neoplasms - metabolism Prostatic Neoplasms - pathology Real time quantitative RT-PCR Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger - metabolism RNA, Neoplasm - analysis Studies Immuno-qPCR Prostate specific antigen Real time quantitative RT-PCR Laser microdissection Human RNA-directed DNA polymerase Microdissection Enzyme Transferases Tumoral marker Real time Protein Polymerase chain reaction Real time quantitative Nucleotidyltransferases Anatomic pathology Messenger RNA RT-PCR Laser Urogenital system Cell Prostate Reverse transcription polymerase chain reaction Quantitative analysis
ISSN:	0046-8177, 1532-8392, 1532-8392
Online-Zugang:	Volltext
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Zusammenfassung:	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 stromal 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.
Bibliographie:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-News-1 content type line 14 ObjectType-Feature-3 ObjectType-Article-1 ObjectType-Feature-2 content type line 23
ISSN:	0046-8177 1532-8392 1532-8392
DOI:	10.1016/j.humpath.2008.02.012