Radiotherapy Sensitization Effect Of Celecoxib On Hypoxia Nasopharyngeal Carcinoma Cell Line CNE-2

Objective:This experiment aimed to investigate radiotherapy sensitization effect the celecoxib on Hypoxia Nasopharyngeal Carcinoma Cell Line CNE-2 and its primary mechanism.Methods:Nasopharyngeal carcinoma cell line CNE-2 were selected as experimental subjects,and cells in logarithmic growth phase were selected for experiment.(1)Tetrazolium salt colorimetric assay(MTT)was used to detect the effect of cobalt chloride(Co Cl2)on the proliferation of Nasopharyngeal Carcinoma Cell Line CNE-2,The appropriate drug concentration was used to induce hypoxia CNE-2 cells.(2)MTT was applied to detect the proliferation effect of celecoxib on Hypoxia Nasopharyngeal Carcinoma Cell Line CNE-2,and the suitable drug concentration was selected;(3)The experiment was divided into Normoxia group,hypoxia group and hypoxia celecoxib group.The colony formation assay was used to detect the radiosensitivity of the Hypoxic Nasopharyngeal Carcinoma Cell Line CNE-2 after celecoxib treatment.(4)The experiment was divided into normoxic control group,Hypoxia control group,hypoxia radiotherapy group,normoxia radiotherapy group and hypoxia celecoxib radiotherapy group.FCM was used to detect the apoptosis of Hypoxic Nasopharyngeal Carcinoma Cell Line CNE-2 with celecoxib combined with radiotherapy Effect of cell cycle.Results:(1)The IC50 values of Co Cl2 treated with CNE-2 cells at 24 h and 48 h were 521.5umol/L and 470.1umol/L,respectively.Within 48 hours,compared with the blank control group,the inhibition rate of CNE-2 cells after 50umol/l and 100umol/l Co Cl2 treatment on CNE-2 cells had no significant difference(P>0.05).There was a significant difference in the inhibition rate of CNE-2 cells between 150umol/l and 200 umol Co Cl2(P<0.05).Co Cl2 at a concentration of 100 umol/l(close to 20% IC50)was chosen for experimental induction of hypoxia.(2)The effects of celecoxib on hypoxic CNE-2 cells 24 h and 48 h showed that celecoxib inhibited the proliferation of hypoxia CNE-2 cells in a time and concentration dependent manner.The IC50 values at 24 h and 48 h were 38.9ug/ml and 36.02ug/ml,respectively.Celecoxib was selected for experiments at a concentration of 10 ug/ml(2 concentrations below the IC50 value).(3)Clone formation experiments showed that after 0,2,4,6Gy irradiation,the survival score(SF)of hypoxia radiotherapy group was higher than that of normoxia radiotherapy group(P<0.05).Survival score(SF)of hypoxia and celecoxib radiotherapy group was lower than that of hypoxia radiotherapy group(P<0.05).To construct single-click multi-target model to fit the cell survival curve,D0,Dq,N in normoxia group,hypoxia group and hypoxemic celecoxib group were(2.82,2.85,2.22),(1.75,2.05,1.35),(1.857,2.051,1.833).Compared with the hypoxic radiotherapy group,radiosensitization ratio(SER)of the hypoxia celecoxib radiotherapy group was 1.28.(4)The result of flow cytometry showed that the distribution of G0-G1 in cell cycle increased gradually compared with normoxia control group,hypoxia control group,hypoxia radiotherapy group,normoxia radiotherapy group and hypoxia celecoxib radiotherapy group,S phase distribution of the cell cycle gradually decreased,the apoptosis rate increased(P<0.05).Conclusion:1.Celecoxib can inhibit the proliferation of Hypoxic Nasopharyngeal Carcinoma Cell Line CNE-2.2.celecoxib can enhance radiosensitivity of Hypoxic Nasopharyngeal Carcinoma Cell Line CNE-2.3.the mechanism may be related to celecoxib inhibiting G0-G1 phase arrest in hypoxic nasopharyngeal carcinoma CNE-2 cells and inducing apoptosis of hypoxia CNE-2 cell.