目的 探讨加热对体外培养的人肝癌细胞株SMMC-7721分泌的细胞因子的影响,以及细胞因子在肿瘤微环境中的作用,为热疗治疗肿瘤提供实验依据.方法 体外培养人肝癌细胞株SMMC-7721至对数生长期,换成无血清培养基,将其放入恒温水浴箱中,42℃恒温加热2 h.将加热后的人肝癌细胞株SMMC-7721放回到37℃,5%CO2的培养箱中进行复温培养,取复温后不同时间(0、24、48、72 h)的人肝癌细胞株SMMC-7721培养上清进行TNF-α、IL-2、IFN-γ和VEGF等抗肿瘤细胞因子的检测,并与未加热的人肝癌细胞株SMMC-7721细胞培养上清进行比较.结果 复温不同时间的人肝癌细胞株SMMC-7721培养上清中TNF-α、IL-2和IFN-γ抗肿瘤细胞因子的分泌浓度均较未加热的人肝癌细胞株SMMC-7721培养上清中上述肿瘤细胞因子的分泌浓度有不同程度的提高,而VEGF的分泌浓度逐渐下降.结论42℃恒温加热2 h可以促进人肝癌细胞株SMMC-7721培养上清中IL-2、IFN-γ、TNF-α等抗肿瘤细胞因子分泌浓度的增加,减少促进肿瘤生长的细胞因子VEGF的分泌浓度.
Objective To investigate the effect of heating on the cytokines secreted by liver cancer cells SMMC-7721,and to study the effect of these cytokines on cell microenvironment,to provide experimental evidence of hyperther-mia for tumor treatment.Method Human liver cancer cell SMMC-7721 were cultured in vitro to the logarithmic growth phase, and then were replaced with a serum-free medium, placed in constant temperature water bath and heated for 2 hours at 42℃.After heating,the human liver cancer cell line SMMC-7721 were placed to cool down to 37℃,and were cultured again in an incubator with 5% CO2by rewarming,the supernatant of the cultures at various time points after re-warming(0,24,48,72 h)were taken to detect TNF-α,IL-2,IFN-γ,VEGF and other cytokines,and were compared with unheated SMMC-7721 cell culture supernatant.Result After rewarming for different duration,the secretion of cytokines TNF-α,IL-2,and IFN-γ in human cancer cells SMMC-7721 were generally increased as compared with unheated cul-tures,while VEGF secretion gradually decreased.Conclusion Constant heating at 42℃for 2 h can promote the secre-tion of cytokines IL-2,IFN-γ,TNF-α,and other anti-tumor cytokines,while reduce the secretion of VEGF that can pro-mote tumor growth.