ILK as a potential marker gene to ascertain specific adenocarcinoma cell mRNA isolation from frozen prostate biopsy tissue sections.
A major technical challenge related to gene expression profiling of tissue samples is the difficulty of procuring selected cell populations from tissues that by nature are heterogeneous, such as prostate tissue. In this study we have examined the expression of integrin-linked kinase (ILK) mRNA in prostate adenocarcinoma cells versus normal prostate epithelial cells in order to determine whether ILK could be used as a reference marker gene for prostate adenocarcinoma cell mRNA isolation. Using laser microdissection (LMD) technology and real-time PCR, together with immunohistochemistry, we have analyzed ILK mRNA expression in epithelial cells isolated from frozen prostate biopsy specimens as well as 4 prostate cell lines (RWPE-1, LNCaP, PC-3 and DU 145) and correlated ILK mRNA expression with ILK protein expression. We demonstrate that quantitative upregulation of ILK mRNA expression in prostate epithelial cells derived from prostate tissue correlated with ILK protein expression and with the histopathology diagnosis of prostate adenocarcinoma. We further show that the level of ILK overexpression was directly influenced by the method used to isolate prostate adenocarcinoma cells (bulk tissue versus LMD dissected cells). These data provide evidence that ILK mRNA is quantitatively upregulated in prostate adenocarcinoma cells versus normal epithelial cells and is therefore a useful internal reference gene marker to evaluate the quality of prostate adenocarcinoma cell derived mRNA used for large scale prostate cancer cDNA gene profiling.
Kieffer
Nelly
N
Laboratoire de Biologie et Physiologie Intégrée LBPI, Université du Luxembourg, L-1511 Luxembourg, Luxembourg. kieffer@cu.lu
Schmitz
Martine
M
Plançon
Sébastien
S
Margue
Christiane
C
Huselstein
Fanny
F
Grignard
Gérard
G
Dippel
Walter
W
Nathan
Michel
M
Giacchi
Sandro
S
Scheiden
René
R
D000230:Adenocarcinoma / Q000737:chemistry / Q000235:genetics*; D014408:Biomarkers, Tumor; D001706:Biopsy; D015153:Blotting, Western; D045744:Cell Line, Tumor; D005629:Frozen Sections; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D042282:Microdissection; D020411:Oligonucleotide Array Sequence Analysis; D011471:Prostatic Neoplasms / Q000737:chemistry / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases / Q000032:analysis / Q000235:genetics*; D011786:Quality Control; D012333:RNA, Messenger / Q000032:analysis*; D000099200:Scaffold Protein ILK
D014408:Biomarkers, Tumor; D017346:Protein Serine-Threonine Kinases; D012333:RNA, Messenger; D000099200:Scaffold Protein ILK