Abstract for presentation at 11th International Congress of Human Genetics

Identification of Differentially Expressed Genes in PA-1 Cells Treated with CR229, a Selective Cyclin-dependent Kinase Inhibitor Generated using Ligand-Based Drug Screening

  • Chul-Hoon Lee, Dept. Medical Genetics, College of Medicine, Hanyang University, Korea
  • Dr Min Kyoung Kim, Dept. Medical Genetics, College of Medicine, Hanyang University, Korea
  • Ha Lim Oh, Dept. Medical Genetics, College of Medicine, Hanyang University, Korea
  • So Hyun Park, Dept. Medical Genetics, College of Medicine, Hanyang University, Korea
  • A Yeon Lee, Dept. Medical Genetics, College of Medicine, Hanyang University, Korea
  • Prof Dr Youl-Hee Cho, Dept. Medical Genetics, College of Medicine, Hanyang University, Korea

    • To generate new scaffold candidates as highly selective and potent cyclin-dependent kinase (CDK) inhibitors, we performed ligand-based drug screening utilizing 3D-pharmacophore conformations of known potent inhibitors. As a result of this 3D-searching, 6-Bromo-2, 3, 4, 9-tetrahydro-carbolin-1-one (CR229) was generated as a hit compound. CR229 is reported here to be a potent inhibitor of CDK2 (IC50: 3 uM) and CDK1 (IC50: 4.9 uM) but had much less effect on other kinases. CR229 treatment was resulted in a decrease in the viability and inducing cell cycle arrest and apoptosis in PA-1 cells than other human cancer cell lines include normal lymphocyte.
    In PA-1 cells, treatment of CR229 resulted in the induction of cell cycle arrest and apoptosis via modulations p53 dependent pathway and inhibition of CDK activities. And together with these data, we compare gene expression between control and CR229 treated PA-1 cells to identify genes that are specifically or predominantly expressed by employing annealing control primer (ACP)-based GeneFishing PCR. Using 120 ACPs, we identified and sequenced 28 differentially expressed genes (DEGs). Among the 28 DEGs, we cloned and sequenced the 10 DEG. These were all exhibited significant sequence similarity with known human genes (98%-100%). Quantitative real time reverse transcription (RT)-polymerase chain reaction (PCR) confirmed that the expression of these genes is altered by treatment of CR229. Of the 10 genes, 2 (HSP90-1, AD33 mitochondrion) were down-regulated and 10 (CIRB, SHMT2, LMNA, MAP1B, GOT1, ZFP513, TOMM20, FLNB) were up-regulated in CR229 treated PA-1 cells. One of the up-regulated genes is Lamin A, which has been known as being a caspase substrate during apoptosis process. One other gene, MAP1B was found to relate with cell division. The genes identified here will provide insights into the cell cycle and apoptosis related roles played by treated CR229 in PA-1 cells.

    Conference Organiser - ICMS Pty Ltd