Targeting p53 in CML Chronic Phase Leukemia Stem Cells

Chronic myelogenous leukemia (CML) is a hematological disease resulting from generation of BCR/ABL oncogene. Resveratrol is an important phytoalexin which is contained in many plants and it has cytotoxic effects in various cancer types. Gene promoter methylation causes loss of tumor suppressor gene function in human cancer. CHD4 (Chromodomain/Helicase/DNAbinding domain) gene which contains a CpG island is a member of the cadherin family. In this study we aimed to evaluate the role of epigenetic modification of resveratrol in cadherin family genes in K562 human CML cell line. K562 cells were treated with 100 mM (IC50 dose) resveratrol depend on time and dosage during 72 hours and cytotoxicity was evaluated by using WST-1 assay. The RT-qPCR is used for gene expression analysis. Gene expression levels were evaluated by using RT Profiler PCR Array. RT-qPCR results showed that, CHD4 gene expression increased 3.45 fold, according to the control cells that untreated with resveratrol. Significant upregulation was found in CHD4 gene expression among the cadherin family genes in resveratrol treated K562 cells. CHD4 is affected by methylation and frequently is silenced in CML. Downregulation of CHD4 gene expression could trigger activation of the Wnt-pathway and cause aberant cell-to-cell interactions. After treatment with the resveratrol in K562 cells, CHD4 gene expression was upregulated. Resveratrol could increase the cadherin family gene expressions and activation of the Wntpathway is blocked in CML. Also, resveratrol could demethylate CpG island in CHD4 promoter region and increase CHD4 gene expression. Our results indicate that resveratrol may act as a demethylation agent on the promoter of tumor supresor genes such as CHD4 and increase the expression of these genes. Therefore, resveratrol could be used as a pioneering anti-tumorigenesis drug in CML. 411 Targeting p53 in CML Chronic Phase Leukemia Stem Cells Luke F. Peterson, Miao-Chia Lo, Yihong Lui, Diane Gianolla, Emilija Mitrikeska, Nicholas J. Donato, Craig N. Johnson, Shaomeng Wang, Jessica Mercer, Moshe Talpaz Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI, USA; Division of Hematology and Oncology, Department of Internal Medicine, University of Michigan, Ann Arbor, MI, USA; Moores UCSD Cancer Center, University of California San Diego, La Jolla, CA, USA; DNA Sequencing and Microarray Core, University of Michigan, Ann Arbor, MI, USA; Department of Pharmacology and Medicinal Chemistry, University of Michigan, Ann Arbor, MI, USA