热化疗诱导舌鳞癌CAL-27细胞免疫原性死亡的实验研究

目的:探讨平阳霉素（pingyangmycin, PYM）、热疗（hyperthermia, HT）及两者联合能否诱导舌鳞癌CAL-27细胞发生免疫原性死亡。方法:对人舌鳞癌细胞CAL-27分别进行PYM、HT及两者联合处理,采用CCK-8实验、Annexin V/PI联合染色、流式细胞术及酶联免疫吸附实验探讨不同处理方式对细胞增殖、凋亡、CRT膜表达及HMGB1分泌的影响。采用SPSS 20.0软件包对数据进行统计学分析。结果:PYM抑制CAL-27细胞活性,且呈浓度依赖性（P<0.05）。PYM及HT均使CRT膜表达率升高;两者联合应用,细胞凋亡、CRT膜表达率及HMGB1分泌均较未处理组、单纯化疗组及单纯HT组升高,差异有统计学意义（P<0.05）。结论:PYM化疗及HT均能诱导舌鳞癌CAL-27细胞CRT由胞内至膜表面转位,并促进HMGB1分泌;两者联合应用,无论在诱导凋亡还是在诱导免疫原性死亡方面,效果优于单纯PYM化疗组。

Study on immunogenic cell death induced by thermo-chemotherapy in CAL-27 cells of tongue squamous cell carcinoma

PURPOSE:To investigate whether pingyangmycin(PYM),hyperthermia(HT)and their combination could induce immunogenic cell death of CAL-27 cells.METHODS:CCK-8 assay,flow cytometry and ELISA assay were used to detect the effect of different treatments on CAL-27 cell proliferation,apoptosis,membrane expression rate of CRT and HMGB1 secretion.SPSS 20.0 software package was used for statistical analysis of the data.RESULTS:PYM inhibited CAL-27 cell activity in a concentration-dependent manner(P<0.05).The membrane expression rate of CRT after PYM treatment and HT treatment were significantly higher than that in the untreated group.After treatment of PYM combined with HT,apoptosis,CRT membrane expression rate and HMGB1 secretion were all increased significantly compared with untreated group,chemotherapy group and HT group(P<0.05).CONCLUSIONS:PYM chemotherapy and HT could induce CRT translocation from intracellular to membrane surface of CAL-27 cells,and combined application of PYM and HT was significantly better than that of PYM alone in inducing apoptosis and immunogenic cell death.