title: Synergistic effect of ponatinib and epigallocatechin-3-gallate induces apoptosis in chronic myeloid leukemia cells through altering expressions of cell cycle regulatory genes.
abstract: OBJECTIVE
  Ponatinib (P) has been used for the treatment of chronic myeloid leukemia (CML) and it is known that inhibition of BCR-ABL fusion protein by ponatinib induces apoptosis of CML cells. Epigallocatechin-3-gallate (EGCG), which is a polyphenol in green tea, induces apoptosis in different types of cancer cells. The purpose of this study was to determine the cytotoxic and apoptotic effects of ponatinib and EGCG combination in K562 CML cell line. This study also aimed to detect alterations of the expression levels of cell cycle-regulation related genes after ponatinib and EGCG combination in K562 CML cell line.
  
  
  METHODS
  The cytotoxic effects of the compounds on K562 cells were determined in a time-and dose-dependent manner by using WST-1 analysis. The combination index (CI) isobologram was used to analyze the data. Apoptotic effects of P-EGCG were defined by flow cytometry and gene expressions were detected by RT-qPCR.
  
  
  RESULTS
  IC50values of ponatinib and EGCG were 87.13 nM and 50μM, respectively. CI value of the P-EGCG was 0.658 and the combination showed synergistic effect (ED90 value: 28.39 nM ponatinib, 117.12 μg/ml EGCG). Ponatinib, EGCG and P-EGCG induced apoptosis compared to control cells. CyclinD1 and CDC25A were downregulated by P-EGCG by 2.49 and 2.63-fold, respectively. TGF-β2 was upregulated by 4.57-fold.
  
  
  CONCLUSIONS
  EGCG possesses cytotoxic and apoptotic properties and may cooperate with the growth inhibiting activity of ponatinib synergistically against CML cells. P-EGCG mediated apoptosis might be associated with upregulation of TGF-β2 gene and downregulation of cyclinD1 and CDC25A genes.
authors.affiliation: Department of Medical Biology, School of Medicine, Ege University, Izmir, Turkey.
authors.forename: Bakiye
authors.initials: B
authors.lastname: Goker
authors.forename: Cansu
authors.initials: C
authors.lastname: Caliskan
authors.forename: Hasan
authors.initials: H
authors.lastname: Onur Caglar
authors.forename: Cagla
authors.initials: C
authors.lastname: Kayabasi
authors.forename: Tugce
authors.initials: T
authors.lastname: Balci
authors.forename: Burcu
authors.initials: B
authors.lastname: Erbaykent Tepedelen
authors.forename: Duygu
authors.initials: D
authors.lastname: Aygunes
authors.forename: Sunde
authors.initials: S
authors.lastname: Yilmaz Susluer
authors.forename: Zeynep
authors.initials: Z
authors.lastname: Mutlu
authors.forename: Nur
authors.initials: N
authors.lastname: Selvi Gunel
authors.forename: Mehmet
authors.initials: M
authors.lastname: Korkmaz
authors.forename: Guray
authors.initials: G
authors.lastname: Saydam
authors.forename: Cumhur
authors.initials: C
authors.lastname: Gunduz
authors.forename: Cigir
authors.initials: C
authors.lastname: Biray Avci
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis; D002392:Catechin / Q000031:analogs & derivatives* / Q000494:pharmacology; D002453:Cell Cycle / Q000235:genetics*; D016044:Fusion Proteins, bcr-abl; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D020014:K562 Cells; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics*; D011724:Pyridazines / Q000494:pharmacology*
chemical_list: D000970:Antineoplastic Agents; D007093:Imidazoles; D011724:Pyridazines; C545373:ponatinib; D002392:Catechin; C045651:epigallocatechin gallate; D016044:Fusion Proteins, bcr-abl