Phenotype-based screening of mechanistically annotated compounds in combination with gene expression and pathway analysis identifies candidate drug targets in a human squamous carcinoma cell model.
The squamous cell carcinoma HeLa cell line and an epithelial cell line hTERT-RPE with a nonmalignant phenotype were interrogated for HeLa cell selectivity in response to 1267 annotated compounds representing 56 pharmacological classes. Selective cytotoxic activity was observed for 14 of these compounds dominated by cyclic adenosine monophosphate (cAMP) selective phosphodiesterase (PDE) inhibitors, which tended to span a representation of the chemical descriptor space of the library. The PDE inhibitors induced delayed cell death with features compatible with classical apoptosis. The PDE inhibitors were largely inactive when tested against a cell line panel consisting of hematological and nonsquamous epithelial phenotypes. In a genome-wide DNA microarray analysis, PDE3A and PDE2A were found to be significantly increased in HeLa cells compared to the other cell lines. The pathway analysis software PathwayAssist was subsequently used to extract a list of proteins and small molecules retrieved from Medline abstracts associated with the hit compounds. The resulting list consisted of major parts of the cAMP-protein kinase A pathway linking to ERK, P38, and AKT. This molecular network may provide a basis for further exploitation of novel candidate targets for the treatment of squamous cell carcinoma.
Fryknäs
Mårten
M
Department of Genetics and Pathology, Uppsala University, S-751 85 Uppsala, Sweden.
Rickardson
Linda
L
Wickström
Malin
M
Dhar
Sumeer
S
Lövborg
Henrik
H
Gullbo
Joachim
J
Nygren
Peter
P
Gustafsson
Mats G
MG
Isaksson
Anders
A
Larsson
Rolf
R
D002294:Carcinoma, Squamous Cell / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism*; D020169:Caspases / Q000378:metabolism; D020650:Combinatorial Chemistry Techniques / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods; D001330:Electronic Data Processing / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D011724:Pyridazines / Q000494:pharmacology; D011799:Quinazolines; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics*
D010726:Phosphodiesterase Inhibitors; D011724:Pyridazines; D011799:Quinazolines; D012333:RNA, Messenger; C042527:imazodan; C040850:quazinone; D010727:Phosphoric Diester Hydrolases; D020169:Caspases; C066436:zardaverine