Table 1.
Prediction results of miRNAs targeting the GDF15 3′ UTR SNP rs1054564.
Fig 1.
Schematic of GDF15 mRNA harboring putative miR-873-5p/miR-1233-3p binding sites and SNP rs1054564 G>C in the 3′ UTR.
Alignment shows the GDF15 3′ UTR variant rs1054564 G>C region with the miRNA hsa-miR-873-5p and hsa-miR-1233-3p motifs. The location of the rs1054564 G>C polymorphism is designated with an arrow in the sequence alignment. Allele G forms a Watson-Crick base-pair with C in the miRNAs (solid line), whereas allele C does not (no line). The seed regions of both miRNAs are designated with an open box. The dots between the base pairs G:U represent GU wobble pairs.
Fig 2.
In silico analysis of the pairing of miR-873-5p and miR-1233-3p to the binding site in the 3′ UTR of GDF15 showing the effect of the minor allele of rs1054564 on the hybrid structures formed between hsa-miR-873-5p/hsa-miR-1233-3p and the GDF15 3′ UTR.
Allele C disrupts the stem part of the typical stem-loop RNA folding structure. The arrow indicates the SNP site in the GDF15 3′ UTR in each folding structure.
Fig 3.
Mimic miRNAs regulate human GDF15 3′ UTR expression in human HEK293T cells.
Results of luciferase reporter activity were analyzed by Student’s t-test and are expressed as mean ± SEMs. (A) Effect of mimic miR-873-5p. (B) Effect of mimic miR-1233-3p. (C) Difference in net suppressive effect of mimic miR-873-5p between rs1054564 alleles G and C. (D) Difference in net suppressive effect of mimic miR-1233-3p between rs1054564 alleles G and C.
Fig 4.
Effect of hsa-miR-873-5p and hsa-miR-1233-3p mimic miRNAs on GDF15 protein expression in human melanoma A2058 cells.
Endogenous GDF15 was suppressed by both mimic miRNAs and restored by miRNA inhibitors. Data represent expression levels relative to those of control samples and indicate the mean of three independent experiments; error bars indicate SE. p-values were determined by a Student’s t-test, and * denotes a p-value < 0.05.