cDNA MICROARRAYS EXPERIMENTAL DESIGN AND GENE EXPRESSION ANALYSIS OF GENISTEIN TREATED HUMAN PROSTATE CANCR CELLS PC-3

  • RAMONA SUHAROSCHI 1Human Nutrition Department. Food Science and Technology. Comparative Oncology Department. University of Agricultural Sciences and Veterinary Medicine of Cluj. Romania. 3-5 Manastur. 3400 Cluj-Napoca.
  • I.R. ROWLAND Northern Ireland Centre for Food and Helath (NICHE). School of Biomedical Sciences. University of Ulster. Cromore Road. Coleraine. N. Ireland. BT52 1SA. UK
  • H. KLOCKER 3Urological Laboratory. Department of Urology. Innsbruck Medical University. Anichstrasse 35. A-6020 Innsbruck. Austria
  • RL. HANCOCK 3Urological Laboratory. Department of Urology. Innsbruck Medical University. Anichstrasse 35. A-6020 Innsbruck. Austria
  • R.S. PARDINI 4Cancer Research Laboratory. Biochemistry Department. School of Medicine. College of Agriculture. University of Nevada. Reno. MS330. 1664 N Virginia St. 89557. Reno. NV. USA
  • AL. BABA University of Agricultural Sciences and Veterinary Medicine of Cluj. Romania. 3-5 Manastur. 3400 Cluj-Napoca
Keywords: PC3, Genistein, cDNA microarrays

Abstract

cDNA microarrays was developed to investigate the relative expression of the corresponding total RNA in prostate cancer cell line PC-3 (AR-) treated with therapeutically doses of Genistein 50uM compared with untreated PC-3 prostate cancer cells. For generating fluorescently labeled cDNA samples to be used in microarray screening we used the SuperScriptTM Indirect cDNA Labeling System (InvitrogenTM). Hybridization procedure was performed overnight using a humidified Corning chamber at 450 C in water bath. The image analysis was performed using GenePix Pro 41121 and ImaGene and for Data analysis we used GeneSight. GeneSpring and MATLAB (MatArray and maanova) software for expression analysis. The gene expression analysis revealed that Genistein at therapeutically and physiologically doses as well caused a differential expression of: “6-phosphofructo-2-kinase/fructose-2. 6-biphosphate 3”. “ATPase. Ca++ transporting. ubiquitous”. “BTG family. member 2”. “H2A histone family. member L”. “H3 histone. family 3A” "UDP-Gal:betaGlcNAc beta 1.4- galactosyltransferase. polypeptide 2". "calcium channel. voltage-dependent. L type. alpha 1C subunit". "calmodulin 3 (phosphorylase kinase. delta)". "caspase 5. apoptosis-related cysteine protease". "chaperonin containing TCP1. subunit 7 (eta)". "general transcription factor IIH. polypeptide 4. 52kDa". "phosphatidylinositol transfer protein. membrane-associated". "polymerase (RNA) II (DNA directed) polypeptide A. 220kDa". "potassium voltage-gated channel. KQT-like subfamily. member 2". "protein tyrosine phosphatase. non-receptor type 3". "protein tyrosine phosphatase. non-receptor type 7". "ras homolog gene family. member N". "transcription elongation factor B (SIII). polypeptide 1 (15kDa. elongin C)". "transcription factor B2. mitochondrial". "tumor necrosis factor. alpha-induced protein 2". Cdc42 guanine nucleotide exchange factor (GEF) 9. DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide 38 acrosomal vesicle protein 1. protocadherin 1 (cadherin-like 1). putative nucleolar RNA helicase. regulator of G-protein signaling 3. ubiquitin specific protease 11. vesicle-associated membrane protein 2 (synaptobrevin 2). We conclude that Genistein can preferentially cause a decrease of some gene involved in apoptosis pathways and an increase in others in human prostate cancer PC-3 cell line. We make a bioinformatics analysis for the identified genes involved in apoptosis induced by Genistein into PC-3 cells. During this analysis we determine the gene sequences. transcripts

Author Biographies

RAMONA SUHAROSCHI, 1Human Nutrition Department. Food Science and Technology. Comparative Oncology Department. University of Agricultural Sciences and Veterinary Medicine of Cluj. Romania. 3-5 Manastur. 3400 Cluj-Napoca.
BIOTECHNOLOGIES
I.R. ROWLAND, Northern Ireland Centre for Food and Helath (NICHE). School of Biomedical Sciences. University of Ulster. Cromore Road. Coleraine. N. Ireland. BT52 1SA. UK
BIOTECHNOLOGIES
H. KLOCKER, 3Urological Laboratory. Department of Urology. Innsbruck Medical University. Anichstrasse 35. A-6020 Innsbruck. Austria
BIOTECHNOLOGIES
RL. HANCOCK, 3Urological Laboratory. Department of Urology. Innsbruck Medical University. Anichstrasse 35. A-6020 Innsbruck. Austria
BIOTECHNOLOGIES
R.S. PARDINI, 4Cancer Research Laboratory. Biochemistry Department. School of Medicine. College of Agriculture. University of Nevada. Reno. MS330. 1664 N Virginia St. 89557. Reno. NV. USA
BIOTECHNOLOGIES
AL. BABA, University of Agricultural Sciences and Veterinary Medicine of Cluj. Romania. 3-5 Manastur. 3400 Cluj-Napoca
BIOTECHNOLOGIES
Published
2009-09-29