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Figure 3 | BMC Biology

Figure 3

From: Mild folate deficiency induces genetic and epigenetic instability and phenotype changes in prostate cancer cells

Figure 3

Mild folate depletion induces genetic damage in prostate cells. (A) High-performance liquid chromatography analysis of nucleotide pools. Results are the average and standard deviation of three independent injections. Values are relative to cells grown in control medium. (B) Single strand breaks generated by treatment with uracil-DNA-glycosidase (UDG) and Exonuclease III are quantified by random oligonucleotide-primed synthesis performed with the Klenow polymerase in the presence of P32-deoxcytidine triphosphate (dCTP; left). The values of dCTP incorporation for all cell lines are the average of five replicates after subtraction of no enzyme treatment control background and presented as an increase over cells grown in control medium. (C) Spectral karyotype analysis reveals that mild folate depletion induced de novo chromosomal aberrations in G100. A representative metaphase from GCtrl (top), which exhibits only marker chromosomes common to both GCtrl and G100 (Table 1) and a representative metaphase from G100 (bottom) which harbours the common t(1;10) translocation along with four marker chromosomes, indicated by arrows, found frequently in G100, but never in GCtrl (Table 1). (D) Distribution of metaphase cells with frequent patterns of marker chromosomes shared between GCtrl and G100 indicates lack of clonal selection in G100. Ten different patterns (A-J) of the five marker chromosomes (a-e; Table 1) were found in two or more metaphase cells. G100 retains significant heterogeneity of patterns of marker chromosomes including two novel patterns (I and J). Pattern A represents marker chromosomes a, b, c and d; pattern B = marker a; pattern C = markers a and e; pattern D = marker b; pattern E = no marker chromosomes; pattern F = markers a, b and c; pattern G = markers b, c and d; pattern H = markers b and c; pattern I = a, b, c, d and e; pattern J = marker e. * = P < 0.05, ** = P < 0.01, *** = P < 0.001.

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