Expression profiles of a human pancreatic cancer cell line upon induction of apoptosis search for modulators in cancer therapy.
We analyzed the differential gene expression in the pancreatic cancer cell line NP-18 upon induction of apoptosis caused by cyclin-dependent kinase inhibition triggered by either overexpression of the tumor suppressor gene p16(INK4A)using an adenoviral construction or incubation with the chemical inhibitors, roscovitine or olomoucine. Screening was performed using cDNA arrays from Clontech that allowed the determination of the expression of 1,176 genes specifically related with cancer. The analysis was carried out using the Atlas Image 2.01 (Clontech) and GeneSpring 4.2 (Silicon Genetics) softwares. Among the differentially expressed genes, we chose for further validation histone deacetylase 1 (HDAC1), von Hippel Lindau and decorin as upregulated genes, and Sp1, hypoxia-inducible factor-1 alpha and DNA primase as downregulated genes. The changes in the expression of these genes to mRNA were validated by quantitative RT-PCR and the final translation into protein by Western blot analysis. Inhibition of HDAC activity, Sp1 binding and DNA primase expression led to an increase in the level of apoptosis, both in parental cells and in doxorubicin-resistant cells. Therefore, these proteins could constitute possible targets to develop modulators in cancer chemotherapy that would increase or restore apoptosis.
Blasco
F
F
Biomedical Research Laboratory, Merck Farma y Química, S.A., Science Park of Barcelona, Barcelona, Spain.
Peñuelas
S
S
Cascalló
M
M
Hernández
J L
JL
Alemany
C
C
Masa
M
M
Calbó
J
J
Soler
M
M
Nicolás
M
M
Pérez-Torras
S
S
Gómez
A
A
Tarrasón
G
G
Noé
V
V
Mazo
A
A
Ciudad
C J
CJ
Piulats
J
J
D000256:Adenoviridae; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D014408:Biomarkers, Tumor / Q000032:analysis*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000032:analysis / Q000235:genetics*; D019915:DNA Primase / Q000032:analysis; D004268:DNA-Binding Proteins / Q000032:analysis; D058575:Decorin; D015536:Down-Regulation; D016326:Extracellular Matrix Proteins; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D019942:Genes, p16 / Q000187:drug effects; D005822:Genetic Vectors; D056284:Histone Deacetylase 1; D006655:Histone Deacetylases / Q000032:analysis; D006801:Humans; D051793:Hypoxia-Inducible Factor 1; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D007701:Kinetin; D009687:Nuclear Proteins / Q000032:analysis; D010190:Pancreatic Neoplasms / Q000737:chemistry* / Q000188:drug therapy; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011509:Proteoglycans / Q000032:analysis; D011687:Purines / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077546:Roscovitine; D014157:Transcription Factors / Q000032:analysis; D025521:Tumor Suppressor Proteins / Q000032:analysis; D044767:Ubiquitin-Protein Ligases / Q000032:analysis; D015854:Up-Regulation; D051794:Von Hippel-Lindau Tumor Suppressor Protein
D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; D019941:Cyclin-Dependent Kinase Inhibitor p16; C546878:DCN protein, human; D004268:DNA-Binding Proteins; D058575:Decorin; D016326:Extracellular Matrix Proteins; C497442:HIF1A protein, human; D051793:Hypoxia-Inducible Factor 1; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D009687:Nuclear Proteins; D047428:Protein Kinase Inhibitors; D011509:Proteoglycans; D011687:Purines; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D000077546:Roscovitine; C090046:olomoucine; D044767:Ubiquitin-Protein Ligases; D051794:Von Hippel-Lindau Tumor Suppressor Protein; D019915:DNA Primase; C110550:HDAC1 protein, human; D056284:Histone Deacetylase 1; D006655:Histone Deacetylases; C497446:VHL protein, human; D007701:Kinetin