Miniaturization of primary porcine urinary bladder epithelial cell cultures and application for gene expression studies after exposure to benzo[a]pyrene.
Benzo[a]pyrene (BaP) is an environmental pollutant used as a key marker substance for polycyclic aromatic hydrocarbons (PAHs). PAHs are believed to play a prominent role in the development of bladder cancer. A test system based on primary porcine urinary bladder epithelial cells (PUBEC) has been utilized as an in vitro model for urinary bladder epithelium. Recently in PUBEC cultures derived from pools of several bladders potent induction of CYP1A1 was detected after BaP treatment. Results from a modified approach using miniaturized PUBEC cultures for the analysis of individual bladder specimens with regard to cell growth and to BaP-mediated induction of CYP1A1 mRNA expression are presented herein. Two types of responses, low and high CYP1A1 induction among individual bladder specimens from eight donor animals, were detected. All of these tissue samples expressed the wild-type genotype of CYP1A1.
Alexander
Michael
M
Department of Structural Biology, Max Planck Institute for Molecular Physiology, Dortmund, Germany.
Behm
Claudia
C
Follmann
Wolfram
W
Kuhlmann
Jurgen
J
D000595:Amino Acid Sequence; D000818:Animals; D001564:Benzo(a)pyrene / Q000633:toxicity*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D019363:Cytochrome P-450 CYP1A1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000737:chemistry; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D008904:Miniaturization / Q000379:methods; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D013552:Swine; D013997:Time Factors; D001743:Urinary Bladder / Q000166:cytology* / Q000187:drug effects
D018076:DNA, Complementary; D012333:RNA, Messenger; D001564:Benzo(a)pyrene; D019363:Cytochrome P-450 CYP1A1