This　study　aimed　to　assess　the　role　of　cyclin-dependent　kinase-like　3　(CDKL3)　in　the　progression　of　non-small　cell　lung　cancer　(NSCLC)　as　well　as　the　underlying　mechanisms.　Western　blot　and　qRT-PCR　were　utilized　to　analyze　CDKL3　expression　in　30　pairs　of　NSCLC　and　paraneoplastic　tissues.　A549　cells　with　CDKL3　knockdown　and　PC9　cells　with　CDKL3　overexpression　were　constructed　by　infecting　cells　with　lentiviruses　expressing　shRNA　of　CDKL3　and　expressing　a　full-length　CDKL3　mRNA,　respectively.　The　CCK-8　assay,　flow　cytometry,　wound　healing　assay,　and　Transwell　assay　were　carried　out　to　detect　cell　viability,　apoptosis,　migration,　and　invasion,　respectively.　Autophagosome　morphology　was　observed　by　electron　microscopy　experiments,　the　expression　of　key　components　of　the　PI3K/Akt/mTOR　pathway　was　examined　via　Western　blot　and　their　mRNA　expression　levels　were　determined.　Besides,　the　stably　infected　NSCLC　cells　with　reduced　expression　or　overexpression　of　CDKL3　were　inoculated　into　the　right-back　flank　of　mice　to　generate　tumors.　The　results　showed　that　CDKL3　expression　was　dramatically　increased　in　NSCLC　tissues.　Moreover,　CDKL3　promoted　the　viability　and　migration　of　NSCLC　cells　by　suppressing　autophagy　in　vitro　and　in　vivo.　In　addition,　CDKL3　might　modulate　PI3K/Akt/mTOR　signaling　in　NSCLC.　Overall,　CDKL3　might　promote　NSCLC　cell　viability　and　metastasis　by　inhibiting　autophagy　and　activating　the　PI3K/Akt/mTOR　signaling　pathway.