title: Clofarabine Has Apoptotic Effect on T47D Breast Cancer Cell Line via P53R2 Gene Expression.
abstract: OBJECTIVE
  Clofarabine, a purine nucleoside analogue and inhibitor of Ribonucleotide Reductase (RR), is used for treatment of leukemia. Clofarabine-induced defect in DNA replication, induces p53 and subsequently P53R2 genes as subunit of RR. clofarabine deregulated P53R2 gene expression leading to the elevated levels of P53R2 which impose resistance to DNA damaging drugs. In this study the apoptotic and cytotoxic effects of clofarabine has been investigated on breast cancer cell line.
  
  
  METHODS
  Cofarabine cytotoxicity on T47D cells has been studied by MTT assay. T47D cells were exposed to the different concentrations of clofarabine for 24, 48 and 72 hours intervals. Relative expression of P53R2 gene has been studied using real-time PCR. Moreover, after treating with clofarabine the apoptotic and necrotic cells were detected using Annexin V and propodium iodide (PI) reagents by flowcytometry technique.
  
  
  RESULTS
  MTT assay results showed that the clofarabine IC50 on T47D cell line were 3 and 2.5µM after 48 and 72 h exposure, respectively. Clofarabine did not show any significant cytotoxic effect after 24 h exposure. The analysis of qRT-PCR showed a significant increase in P53R2 gene expression in treated cells with both 2.5 and 3 μM doses and also, the results of flowcytometry revealed 26.91 and 74.46 percent apoptosis induction in 48 and 72h treatments respectively in comparison to the control groups.
  
  
  CONCLUSIONS
  Our results showed that apoptotic and cytotoxic effects of clofarabine on T47D cell line were in time and dose dependent manner; therefore it could be considered a new candidate in breast cancer therapy.
authors.affiliation: Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.; Department of Clinical Biochemistry, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Rahmati-Yamchi
authors.affiliation: Department of Clinical Biochemistry, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Nosratollah
authors.initials: N
authors.lastname: Zarghami
authors.affiliation: Stem Cell Research center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Hojjatollah
authors.initials: H
authors.lastname: Nozad Charoudeh
authors.affiliation: Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.; Department of Clinical Biochemistry, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Yasin
authors.initials: Y
authors.lastname: Ahmadi
authors.affiliation: Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Behzad
authors.initials: B
authors.lastname: Baradaran
authors.affiliation: Department of Biology, Faculty of Natural Science, University of Tabriz, Tabriz, Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Khalaj-Kondori
authors.affiliation: Liver and Gastrointestinal disease research center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Morteza
authors.initials: M
authors.lastname: Milani
authors.affiliation: Liver and Gastrointestinal disease research center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Abolfazl
authors.initials: A
authors.lastname: Akbarzadeh
authors.affiliation: Department of Clinical Biochemistry, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Maghsud
authors.initials: M
authors.lastname: Shaker
authors.affiliation: Department of Medical biotechnology, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Pourhassan-Moghaddam
keywords: Apoptosis; Breast Cancer; DNA damage; P53R2
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