PMID- 11595508
OWN - NLM
STAT- MEDLINE
DCOM- 20011205
LR  - 20190901
IS  - 0960-0760 (Print)
IS  - 0960-0760 (Linking)
VI  - 78
IP  - 3
DP  - 2001 Sep
TI  - Stable expression of human 5alpha-reductase type II in COS1 cells due to 
      chromosomal gene integration: a novel tool for inhibitor identification.
PG  - 275-84
AB  - Inhibitors of human 5alpha-reductase type II are promising drug candidates for 
      the treatment of benign prostatic hyperplasia which is associated with high 
      prostatic DHT levels. In this study we describe the evaluation of potential 
      inhibitors in a new cell assay. First a plasmid (pRcCMV-5alphaII) for the 
      expression of human 5alpha-reductase type II was constructed by the use of the 
      vector pRcCMV and transfected into the African green monkey fibroblast-like cell 
      line COS1. By selection with G418 sulfate, ten COS1 single cell clones were 
      obtained of which three stably exhibited high 5alpha-reductase activity. One 
      single cell clone (COS1-5alphaIIST) was selected for further investigations. By 
      Southern blot analysis, fluorescence in situ hybridization (FISH) and comparative 
      PCR experiments it turned out that the expression plasmid pRcCMV-5alphaII has 
      been integrated into the chromosome, resulting in a long-term stable expression 
      of the foreign 5alpha-reductase gene. The newly established cell line was used 
      for testing novel compounds on their inhibitory effect on human 5alpha-reductase 
      type II. Using this whole cell assay, inhibitors with IC(50) values in the 
      nanomolar range could be identified.
FAU - Reichert, W
AU  - Reichert W
AD  - FR 8.5 Pharmaceutical and Medicinal Chemistry, University of the Saarland, P.O. 
      Box 151150, D-66041, Saarbrucken, Germany.
FAU - Michel, A
AU  - Michel A
FAU - Hartmann, R W
AU  - Hartmann RW
FAU - Jose, J
AU  - Jose J
LA  - eng
PT  - Journal Article
PL  - England
TA  - J Steroid Biochem Mol Biol
JT  - The Journal of steroid biochemistry and molecular biology
JID - 9015483
RN  - 0 (DNA Primers)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Isoenzymes)
RN  - 0 (Recombinant Proteins)
RN  - 08J2K08A3Y (Dihydrotestosterone)
RN  - 409J2J96VR (Androstenedione)
RN  - EC 1.- (Oxidoreductases)
RN  - EC 1.3.1.22 (Cholestenone 5 alpha-Reductase)
SB  - IM
MH  - Androstenedione/metabolism
MH  - Animals
MH  - Base Sequence
MH  - COS Cells
MH  - Cholestenone 5 alpha-Reductase
MH  - Chromosome Mapping
MH  - DNA Primers/genetics
MH  - Dihydrotestosterone/metabolism
MH  - Drug Evaluation, Preclinical
MH  - Enzyme Inhibitors/pharmacology
MH  - Gene Expression
MH  - Humans
MH  - Isoenzymes/antagonists & inhibitors/genetics
MH  - Male
MH  - Oxidoreductases/antagonists & inhibitors/*genetics
MH  - Plasmids/genetics
MH  - Prostatic Hyperplasia/drug therapy/metabolism
MH  - Recombinant Proteins/antagonists & inhibitors/genetics
MH  - Transfection
EDAT- 2001/10/12 10:00
MHDA- 2002/01/05 10:01
CRDT- 2001/10/12 10:00
PHST- 2001/10/12 10:00 [pubmed]
PHST- 2002/01/05 10:01 [medline]
PHST- 2001/10/12 10:00 [entrez]
AID - S0960-0760(01)00092-9 [pii]
AID - 10.1016/s0960-0760(01)00092-9 [doi]
PST - ppublish
SO  - J Steroid Biochem Mol Biol. 2001 Sep;78(3):275-84. doi: 
      10.1016/s0960-0760(01)00092-9.