Pyruvate dehydrogenase kinase 4 (PDK4) was upregulated in gastric cancer (GC) tissues and cells, and PDK4 was identified as a direct target of miR‐5683. (A) The color of the culture medium in SGC7901. (B) Target gene searched on TargetScan, miRDB, starBase, Tarbase, and RNA22 prediction websites. (C) Relative expression of PDK1, PDK2, PDK3, and PDK4 after miR‐5683 downregulation. (D) The association of PDK4 with GC stage using GEPIA database. (E, F) Kaplan‐Meier analysis of the association of PDK4 with overall survival (E) or recurrence‐free survival (F) in patients with GC using GEPIA database. (G) PDK4 expression levels determined in 70 pairs of human GC tissues and adjacent normal tissues by qRT‐PCR. (H, I) The potential miR‐5683 binding site at the 3'‐UTR of PDK4 mRNA was computationally predicted by TargetScan. Luciferase activity was analyzed in cells co‐transfected with miR‐5683‐mimics or negative control and pGL3‐PDK4 or pGL3‐PDK4‐mut. (J) PDK4 protein expression in GC specimens and adjacent normal tissues determined by immunohistochemistry staining. (K) PDK4 protein expression was analyzed by western blotting in six pairs of GC tissues. (L) A negative correlation between the expression levels of miR‐5683 and PDK4 in GC specimens (P < .05). (M) PDK4 protein expression in GC cells transfected with lentiviruses expressing miR‐5683‐mimics and miR‐5683‐inhibitor. (N) Effects of changes in miR‐5683 expression on PDK4 mRNA expression in GC cells analyzed by qRT‐PCR. *p < .05, **p < .01, ***p < .001.