Objective To establish the paclitaxel-resistant gastric cancer cell(HGC-27/PTX) and to investigate the changes of characteristics before and after resistance,as well as the possible resistant mechanisms.Methods The paclitaxel-resistant gastric cancer cell HGC-27/PTX was established by increasing paclitaxel dose gradually and intermittently.The IC50 (50% inhibitory concentration) and cell cycle were determined by CCK-8 assay and flow cytometry,respectively.The differentially expressed genes (DEGs) and signaling pathways were analyzed using RNAseq.Results The establishment of HGC-27/PTX cells lasted 9 months,and the sensitivity of paclitaxel of HGC-27/PTX cells was significantly lower than parental cells (P<0.05).Compared to parental cells,the morphology of HGC-27/PTX cells was slightly different,and the proportion of S and G2/M phase was obviously increased (P<0.01).A total of 274 DEGs were identified between the resistant and parental cells with 130 genes up-regulated and 144 genes down-regulated.DEGs were significantly enriched in extracellular matrix (ECM)-receptor interaction(P<0.001) and PI3K-Akt signaling pathways (P<0.05),which could provide evidences for reversing paclitaxel resistance.Conclusions The paclitaxel-resistant gastric cancer cells HGC-27/PTX was established with stable culturein vitro,which provides an ideal model for future study on the mechanism of drug resistance.