Reversal Effect Of Kanglaite Injection On Drug Resistance Of Human Nasopharyngeal Carcinoma Cell Line CNE2/DDP

Objective: to study whether Kanglaite injection can reverse the drug resistance of human nasopharyngeal carcinoma CNE2/DDP cells and its mechanism,in order to provide experimental basis for improving the chemosensitivity of nasopharyngeal carcinoma to develop new drugs。Materials and methods: human nasopharyngeal carcinoma CNE2 and CNE2/DDP cells were used as experimental objects,and the morphological differences between drug-resistant and non-drug-resistant strains were observed by inverted microscope.The effects of different concentrations of cisplatin(0,0.35ug/m L,0.7ug/m L,1.4ug/m L,2.8ug/m L,5.6ug/m L,11.2ug/m L)and Kanglaite injection(0,0.125g/L,0.25g/L,0.5g/L)on the proliferation of CNE2 cells were detected by CCK8.CNE2/DDP cells were treated with different concentrations of cisplatin(0,0.125g/L,0.25g/L,0.5g/L)for 48 h.After CNE2/DDP48 h was treated with Kanglaite injection,which had no obvious inhibitory effect on CNE2/DDP cells,the expression of P-glycoprotein(P-gp)and Cleaved Caspase-3 in CNE2/DDP cells was detected by Western blot.Results: 1.Under inverted microscope,CNE2/DDP cells showed irregular morphology,relatively slow growth,larger volume than CNE2 cells,clear granular substances in the cytoplasm,multinucleated giant cells were easy to see,and their adhesion ability was weaker than that of CNE2 cells,that is,the time of trypsin digestion was shorter than that of CNE2 cells.2.After CNE2/DDP and CNE2 cells were treated with different concentrations of cisplatin,CCK8 results showed that the cell survival rate of cisplatin-resistant human nasopharyngeal carcinoma cell line CNE2/DDP was higher than that of CNE2.The IC50 value of CNE2 cells was 1.610 ±0.1362 and the drug resistance index of CNE2/DDP cells was 13.343 ±1.5404.Kanglaite injection at different concentrations inhibited the proliferation of human nasopharyngeal cancer cells CNE2 and CNE2/DDP cells in a concentration-dependent manner.Among them,0.125g/L Kanglaite injection had no cytotoxic effect on CNE2/DDP cells(inhibition rate < 5%),while 0.25g/L Kanglaite injection had weak cytotoxicity on CNE2/DDP cells(inhibition rate < 10%).CNE2/DDP cells were treated with Kanglaite injection of 0.125 and 0.25g/L.The reversal ratios of cisplatin resistance of 0.125g/L and 0.25g/L Kanglaite injection to human nasopharyngeal cancer cell CNE2/DDP cells were1.29,1.94,respectively.The results of Western blotting showed that the expression of P-gp protein decreased and the expression of Cleaved Caspase-3 protein increased in 0.125g/L Kanglaite injection group and0.25g/L Kanglaite injection group.Compared with the control group,the expression of P-gp protein in 0.125g/L Kanglaite injection group decreased slightly,while the expression of Cleaved Caspase-3 protein increased slightly,but the difference was not statistically significant.The expression of P-gp protein in the 0.25g/L Kanglaite injection group was significantly lower than that in the control group,and the difference was statistically significant(P < 0.05).The expression of Cleaved Caspase-3protein in the Kanglaite injection group was significantly higher than that in the control group(P < 0.01).Conclusion: 1.The drug resistance index of human nasopharyngeal carcinoma CNE2/DDP cells was 8.2814,which proved that the cells had drug resistance.2.Different concentrations of Kanglaite injection inhibited the proliferation of human nasopharyngeal carcinoma cells CNE2 and CNE2/DDP in a concentration-dependent manner.3.Kanglaite injection can reverse the drug resistance of human nasopharyngeal carcinoma CNE2/DDP cells.4.The mechanism of Kanglaite injection reversing the drug resistance of human nasopharyngeal carcinoma CNE2/DDP cells may be related to that Kanglaite injection can inhibit the expression of P-gp protein and increase the expression of Cleaved Caspase-3 protein,that is,it can reverse drug resistance by regulating the expression of multidrug resistance genes and promoting apoptosis.