Preliminary Study On The Expression Of CRT,HSP70,HMGB1 And ATP In Tongue Cancer Cal27 Cells By Anlotinib Hydrochloride And Paclitaxel

Objective: To investigate the effects of anlotinib hydrochloride and paclitaxel on the expression of immunogenic death-related molecules CRT,HSP70,HMGB1 and ATP in tongue cancer Cal27 cells.Methods: 1.Tongue cancer Cal27 cells were selected as the research object,the optimal concentration and action time of Al3818 and PTX were detected using CCK-8 method;2.The cells were divided into blank control group,AL3818 group,PTX group,and AL3818+PTX group;3.The apoptosis of cells in different experimental groups were detected using flow cytometry;4.The expression of CRT and HSP70 in Cal27 cells were detected using immunofluorescence and flow cytometry;5.Enzyme-linked immunoassay was adopted to detect the expression of HMGB1 in Cal27 cell supernatant;6.ATP luminescence kit was used to detect the expression of ATP in Cal27 cell supernatant;7.The above experiments were repeated three times,and the results were statistically analyzed using SPSS 18.0 software,and the results were expressed as mean ± standard deviation.The independent samples t test was adopted for the comparison between two groups and single factor one-factor variance analysis was adopted for multi-group comparison;P <0.05 was considered statistically different.Results: 1.CCK-8 results showed that 24 h was the optimal time for the effect of AL3818 and PTX on Cal27 cells,6.254μmol/L and 1.718μmol/L was the optimal concentration,respectively;2.Flow cytometry experiment results showed that AL3818 and PTX could induce Cal27 cell apoptosis,which had statistical differences(P<0.05)compared with the control group.AL3818 + PTX group had the highest apoptosis rate,which had statistical differences(P<0.05)compared with control group,Al3818 group,and PTX group;3.Results of immunofluorescence,flow cytometry,enzyme-linked immunization experiment,and ATP luminescent kit experiment showed that AL3818 and PTX could induce the exposure of CRT,HSP70,HMGB1,and ATP in Cal27 cells,which had statistical differences(P<0.05)compared with control group.AL3818+PTX group had the optimal effect,which had statistical differences(P<0.05)compared with control group,AL3818 group,and PTX group.Conclusion: AL3818 and PTX could induce immunogenic death of tongue cancer,and this process was related to the exposure of immune-related molecules CRT,HSP70,HMGB1,and ATP.The immune effect of AL3818 combined with PTX on tongue cancer was better than the separate effect of AL3818 and PTX.