Fig 1: MiR‐4664‐3p can promote the cell cycle and target‐downregulate CDK2AP2 expression, which is an essential component for CDK2AP1 function. (a) TargetScan analysis predicted that the CDK2AP2 mRNA‐3′‐UTR is targeted by miR‐4664‐3p. CDK2AP2 mRNA 3′‐UTR or mutated CDK2AP2 mRNA 3′‐UTR was cloned into psiCHECK2. (b) Dual‐luciferase gene reporter assay of miRNA cotransfection with psiCHECK2 containing CDK2AP2 mRNA 3′‐UTR or mutated CDK2AP2 mRNA 3′‐UTR; analysis of variance (ANOVA) test. (c) Quantitative reverse transcription‐polymerase chain reaction (qRT‐PCR) and western blot detection of CDK2AP2 in miR‐4664‐3p, miR‐4664‐3p inhibitor, or scramble‐treated A549 cells. (d) Western blot detection of the p53/p21/p27/CDK2/cyclin E1 axis in miR‐4664‐3p, miR‐4664‐3p inhibitor, or scramble‐treated A549 cells. (e) Cell cycle analyzed by flow cytometry in miR‐4664‐3p mimic, miR‐4664‐3p inhibitor, or scrambled‐treated A549 cells; student's t‐test. (f) CDK2AP2 expression in xenografts analyzed by immunohistochemistry (IHC). (g) Western blot detection of the CDK2AP2/p53/p21/p27/CDK2/cyclin E1 axis in xenografts. (h) Co‐immunoprecipitation (Co‐IP) assay detecting the interaction between CDK2AP2 and CDK2AP1 in Flag‐pcDNA‐CDK2AP1 transfected cells. (i) 5‐ethynyl‐2′‐deoxyuridine (EdU) assay of A549 cells with cotransfection of pcDNA3.1 + NC, pcDNA‐CDK2AP1 + NC, pcDNA3.1 + si‐CDK2AP2 or pcDNA‐CDK2AP1+ si‐CDK2AP2. (j) Cell counting kit‐8 (CCK‐8) assay of A549 in 72 h post‐transfection of pcDNA3.1 + NC, pcDNA‐CDK2AP1 + NC, pcDNA3.1 + si‐CDK2AP2 or pcDNA‐CDK2AP1+ si‐CDK2AP2; analysis of variance (ANOVA) test. Data are expressed as mean ± SD for triplicate experiments. ***p < 0.001, **p < 0.01, *p < 0.05.