BACKGROUND: In this study, the effect of clofarabine, a new generation 2'-deoxyadenosine analogue, on promoter methylation and transcriptional activity of selected genes (PTEN, APC, RARB2, ZAP70) in K562 cells was assessed. MATERIALS AND METHODS: Promoter methylation was estimated using methylation-sensitive restriction analysis. The mRNA level of the genes was measured with real-time PCR. RESULTS: The inhibitory cytostatic index (IG(50)) for clofarabine in K562 cells cultured for 72 (or 96) h was 8 nM. The drug (20 nM) caused: (i) potent diminution in methylation of PTEN promoter, moderate methylation reduction of APC and RARB2 promoters, and complete methylation of ZAP70 promoter; (ii) significant stimulation of PTEN, APC, RARB, and p21 mRNA expression and (iii) decline in mRNA level of ZAP70 and DNMT1 genes. CONCLUSION: The results indicated that clofarabine is involved in epigenetic regulation of transcriptional activity of the tested tumour suppressor genes and genes encoding proteins involved in DNA methylation process.
BACKGROUND: In this study, the effect of clofarabine, a new generation 2'-deoxyadenosine analogue, on promoter methylation and transcriptional activity of selected genes (PTEN, APC, RARB2, ZAP70) in K562 cells was assessed. MATERIALS AND METHODS: Promoter methylation was estimated using methylation-sensitive restriction analysis. The mRNA level of the genes was measured with real-time PCR. RESULTS: The inhibitory cytostatic index (IG(50)) for clofarabine in K562 cells cultured for 72 (or 96) h was 8 nM. The drug (20 nM) caused: (i) potent diminution in methylation of PTEN promoter, moderate methylation reduction of APC and RARB2 promoters, and complete methylation of ZAP70 promoter; (ii) significant stimulation of PTEN, APC, RARB, and p21 mRNA expression and (iii) decline in mRNA level of ZAP70 and DNMT1 genes. CONCLUSION: The results indicated that clofarabine is involved in epigenetic regulation of transcriptional activity of the tested tumour suppressor genes and genes encoding proteins involved in DNA methylation process.
Authors: Nikolas Herold; Sean G Rudd; Linda Ljungblad; Kumar Sanjiv; Ida Hed Myrberg; Cynthia B J Paulin; Yaser Heshmati; Anna Hagenkort; Juliane Kutzner; Brent D G Page; José M Calderón-Montaño; Olga Loseva; Ann-Sofie Jemth; Lorenzo Bulli; Hanna Axelsson; Bianca Tesi; Nicholas C K Valerie; Andreas Höglund; Julia Bladh; Elisée Wiita; Mikael Sundin; Michael Uhlin; Georgios Rassidakis; Mats Heyman; Katja Pokrovskaja Tamm; Ulrika Warpman-Berglund; Julian Walfridsson; Sören Lehmann; Dan Grandér; Thomas Lundbäck; Per Kogner; Jan-Inge Henter; Thomas Helleday; Torsten Schaller Journal: Nat Med Date: 2017-01-09 Impact factor: 53.440