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Fig. 2 | Molecular Cancer

Fig. 2

From: DNA methylation-associated dysregulation of transfer RNA expression in human cancer

Fig. 2

tDNA methylation is associated with different overall survival in TCGA cohorts. A Dot plots summarizing the logrank tests (left) and univariate Cox regression models (right) used to compare the overall survival of patients according to methylation status of the 71 tDNAs. Cases that are significantly associated with different prognosis are represented in large-sized bullets. Yellow and blue represent favorable and unfavorable prognosis according to univariate Cox regression analyses (right), respectively. HR, hazard ratio. B Kaplan-Meier curves show that tRNA-Arg-TCT-4-1 hypomethylation is associated with a shorter overall survival in KIRP (top) and UCEC (below) TCGA cohorts. HR, hazard ratio; CI, confidence interval. p-values correspond to logrank tests. C Bisulfite genomic sequencing confirms tRNA-Arg-TCT-4-1 hypomethylation in HEC1 cell line. The tDNA sequence is indicated with a blue bracket. The orange rectangles correspond to the A and B boxes of the tDNA. The TSS is marked with a black arrow. CpG dinucleotides are represented as short vertical lines, and their methylation status is denoted with black (methylated) or white (unmethylated) squares. The CpG represented in the HM450 microarray is marked with a red asterisk. D qRT-PCR exposes higher tRNA-Arg-TCT-4-1 levels in HEC1 compared to DND41 and SW48 cells. Data represent the mean ± SD of biological triplicates analyzed by an unpaired two-tailed Student’s t-test. *** p < 0.001. E Cell cycle analysis reveals an accumulation of tRNA-Arg-TCT-4-1 knockout HEC1 cells in G0/G1 phase. Data shown represent the mean ± SD of biological triplicates analyzed by unpaired two-tailed Student’s t-test. * p < 0.05. F SRB assay shows a reduced growth of HEC1 knockout cells. Data at each time points are the mean ± SD of four biological replicates. Statistical differences were determined using an unpaired two-tailed Student’s t-test at the 144 h final time point. * p < 0.05. G Transwell assay shows a reduced migration of tRNA-Arg-TCT-4-1-silenced HEC1 cells. Data represent the mean ± SD of biological triplicates analyzed by an unpaired two-tailed Student’s t-test. *** p < 0.001. Representative images of the Transwell insert membranes are shown.

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