Abstract: Objective To establish radiation-resistant nasopharyngeal carcinoma cell subline CNE-2S and reveal the relationship between expression levels of SHP-1/p21/CDK6/Cyclin D1 and radiation-resistance in nasopharyngeal carcinoma cells. Methods Radiation-resistant nasopharyngeal carcinoma cell subline was set up by X-ray hypofractionted radiotherapy.The differences on morphology and growth kinetics between different passengers were observed.The cell cycles of different passengers were analyzed.The protein levels in SHP-1/p21/CDK6/Cyclin D1 signaling pathway were examined in various clones of cells. Results The X-ray hypofractionted radiotherapy could build radioresistant subline of nasopharyngeal carcinoma.Meanwhile,the percentage of CNE-2S1 cells in S phase was significantly increased than that of CNE-2 cells,G₁ phase cells were decreased significantly,G₂-M phase cells had no obvious change.In addition,protein levels of SHP-1,CDK6,and CylinD1 in CNE-2S1 cells were significantly increased,but the expression of p21 was significantly reduced than that of CNE-2 cells. Conclusion SHP-1-p21-CyclinD1-CDK6 pathway may have a certain role in radiation sensitivity and cell cycle distribution of nasopharyngeal carcinoma.