Paclitaxel　(PTX),　a　tubulin-binding　agent,　is　widely　used　and　has　shown　good　efficacy　in　the　initial　period　of　treatment　for　non-small　cell　lung　cancer　(NSCLC).　However,　the　relatively　rapid　acquisition　of　resistance　to　PTX　treatments　that　is　observed　in　virtually　all　cases　significantly　limits　its　utility　and　remains　a　substantial　challenge　to　the　clinical　management　of　NSCLC.　The　aim　of　this　study　was　to　identify　candidate　genes　and　mechanisms　that　might　mediate　acquired　paclitaxel　resistance.　In　this　work,　we　established　paclitaxel-resistant　cells　(A549-T)　from　parental　cell　lines　by　step-dose　selection　in vitro.　Using　methylation　chip　analysis　and　transcriptome　sequencing,　43,426　differentially　methylated　genes　and　2,870　differentially　expressed　genes　are　identified.　Six　genes　(KANK1,　ALDH3A1,　GALNT14,　PIK3R3,　LRG1,　WEE2),　which　may　be　related　to　paclitaxel　resistance　in　lung　adenocarcinoma,　were　identified.　Among　these　genes,　KANK1　exhibited　significant　differences　in　methylation　and　expression　between　cell　lines.　Since　KANK1　plays　an　important　role　in　the　development　of　renal　cancer　and　gastric　cancer,　we　hypothesised　that　it　may　also　play　a　role　in　acquired　resistance　in　lung　adenocarcinoma.　Transient　transfection　of　SiKANK1　significantly　reduced　the　expression　of　KANK1,　reducing　apoptosis,　increasing　cell　migration,　and　enhancing　the　tolerance　of　A549　cells　to　paclitaxel.　KANK1　acts　as　a　tumour　suppressor　gene,　mediating　the　resistance　of　lung　adenocarcinoma　A549　to　paclitaxel.　The　reduction　of　KANK1　expression　can　increase　the　paclitaxel　resistance　of　non-small　cell　lung　cancer　and　increase　the　difficulty　of　clinical　treatment.　©　2020,　©　2020　The　Author(s).　Published　by　Informa　UK　Limited,　trading　as　Taylor　&　Francis　Group.