Journal: Leukemia

Article Title: Expression of nucleoside-metabolizing enzymes in myelodysplastic syndromes and modulation of response to azacitidine

doi: 10.1038/leu.2013.330

Figure Lengend Snippet: Expression of genes implicated in the metabolism of AZA. ( a ) Expression of UCK1 , UCK2 , DCK , hENT1 , RRMI and RRM2 (relative to GAPDH ) in patients with MDS ( n =57) before AZA treatment. ( b ) Expression of UCK1 (relative to GAPDH ) in patients without response to AZA (SD or PD) vs patients with response to AZA (CR, PR, HI and mCR). In both plots, the thick middle line represents the median expression of the different genes, and the top and bottom edges of the box represent the 75th and 25th percentiles, respectively. The outliers are also represented by open and filled circles and asterisks.

Article Snippet: Specific oligonucleotides and TaqMan probes were available from Applied Biosystems (catalog no: UCK1: Hs01075618_m1; UCK2 : Hs:00367072_m1, DCK : Hs00176127_m1, hENT1 : Hs01085706_m, RRM1 : Hs01040698_m1, RRM2 : Hs01072069_g1).

Techniques: Expressing

Journal: Leukemia

Article Title: Expression of nucleoside-metabolizing enzymes in myelodysplastic syndromes and modulation of response to azacitidine

doi: 10.1038/leu.2013.330

Figure Lengend Snippet: Molecular alterations of UCK1 in MDS patients. ( a ) Representative examples of methylation-specific PCR analysis of UCK1 in seven MDS cases without response to AZA and six cases of MDS patients with response to AZA. C(+), DNA universally methylated; C(−), healthy individual; blank, control without added DNA; U, unmethylated alleles; and M, methylated. ( b ) UCK1 gene locus and the sequenced regions. Five PCR were designed to sequence the complete coding DNA sequence (Exon 1, Exon 2, Exon 3-5, Exon 6 and Exon 7) for all the isoforms of the UCK1 protein. A total of seven common genetic variants, indicated by their reference number, were found to be polymorphic in at least one subject. ( c ) UCK1 expression levels in different genotype groups. Gene expression values were compared with the genotypes of the genetic variants identified. In the plots, the thick middle line represents the median expression of the different genes, and the top and bottom edges of the box represent the 75th and 25th percentiles, respectively. The outliers are represented by asterisks.

Article Snippet: Specific oligonucleotides and TaqMan probes were available from Applied Biosystems (catalog no: UCK1: Hs01075618_m1; UCK2 : Hs:00367072_m1, DCK : Hs00176127_m1, hENT1 : Hs01085706_m, RRM1 : Hs01040698_m1, RRM2 : Hs01072069_g1).

Techniques: Methylation, Control, Sequencing, Expressing, Gene Expression

Journal: Leukemia

Article Title: Expression of nucleoside-metabolizing enzymes in myelodysplastic syndromes and modulation of response to azacitidine

doi: 10.1038/leu.2013.330

Figure Lengend Snippet: Silencing of the UCK1 gene. Effect of siUCK1-1 on the expression of the UCK1 gene (relative to GAPDH ) was evaluated using quantitative PCR ( a ) and western blotting analysis ( b ). Downregulation of the UCK1 mRNA by siUCK1-1 was observed in non-transfected compared with mock transfected control cells, both not treated (* P =0.046) and treated with AZA (** P =0.021) ( a ). Mean of two different experiments is shown. UCK1 protein expression was equally diminished in K562 cells exposed to siUCK1-1 ( b ). ( c ) Apoptosis (Annexin V-FITC (fluorescein isothiocyanate) and PI staining) was measured in K562 cells cultured with SRC, TKO and siUCK1-1, without AZA and with 24-h AZA. AZA induced an increase in Annexin V-positive cells, but apoptosis was blunted in K562 cells when siUCK1-1 was present and UCK1 , as well as UCK1 protein, were downexpressed (* P =0.0015, siUCK1-1 transfected vs control culture).

Article Snippet: Specific oligonucleotides and TaqMan probes were available from Applied Biosystems (catalog no: UCK1: Hs01075618_m1; UCK2 : Hs:00367072_m1, DCK : Hs00176127_m1, hENT1 : Hs01085706_m, RRM1 : Hs01040698_m1, RRM2 : Hs01072069_g1).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Transfection, Control, Staining, Cell Culture

Journal: Leukemia

Article Title: Expression of nucleoside-metabolizing enzymes in myelodysplastic syndromes and modulation of response to azacitidine

doi: 10.1038/leu.2013.330

Figure Lengend Snippet: OS by UCK1 expression. Kaplan–Meier analysis. Solid lines indicate OS of patients with expression of UCK1 above the median; dashed lines indicate OS of patients with expression of UCK1 below the median.

Article Snippet: Specific oligonucleotides and TaqMan probes were available from Applied Biosystems (catalog no: UCK1: Hs01075618_m1; UCK2 : Hs:00367072_m1, DCK : Hs00176127_m1, hENT1 : Hs01085706_m, RRM1 : Hs01040698_m1, RRM2 : Hs01072069_g1).

Techniques: Expressing