融合蛋白G3G6和HST1致敏DC疫苗抗黑色素瘤的药效学研究

Pharmacological effects of anti-melanoma DC vaccine sensitized by fusion proteins of G3G6 and HST1

探究促性腺激素释放激素(GnRH)与胃泌素释放肽(GRP)偶联的融合蛋白(G3G6)和热休克蛋白65(HSP65)与六跨膜前列腺上皮抗原(STEAP1)偶联的融合蛋白(HST1)致敏树突状细胞(DC)的效果和致敏后DC对B16F10黑色素瘤的抑制杀伤作用。利用实验室现存工程菌表达融合蛋白G3G6和HST1,按未致敏DC(US-DC)组,G3G6融合蛋白致敏DC(G3G6-DC)组,HST1融合蛋白致敏DC(HST1-DC)组和G3G6及HST1联合致敏DC(GH-DC)组致敏小鼠骨髓来源的DC分化成熟,获得融合蛋白致敏的相应DC疫苗。将B16F10黑色素瘤细胞以1×10~6个/只移植于C57BL/6J雄性小鼠构建黑色素瘤模型,DC疫苗免疫治疗,体内外实验探究DC疫苗的抗肿瘤药效。流式细胞术分析证明,融合蛋白有效刺激DC分化成熟;动物实验显示,与US-DC组相比,G3G6-DC黑色素瘤抑制率为35.75%,HST1-DC组为34.03%,GH-DC组为55.74%。研究结果初步证明,G3G6-DC和HST1-DC均可有效抑制黑色素瘤B16F10细胞小鼠移植瘤的生长(P<0.05),且联合用药优于单独用药(P<0.01)。

This study aimed to investigate the effects of fusion proteins GnRH-GRP (G3G6) and HSP65-STEAP1 (HST1) on dendritic cells (DC) and the sensitization of DCs to B16F10 melanoma.The fusion proteins G3G6 and HST1 were obtained using the previous engineering strains in our laboratory.Group by unsensitized DC (US-DC),the G3G6 fusion protein sensitized DC,the HST1 fusion protein sensitized DC (HST1-DC) and the combined sensitized DC (GH-DC),the mouse bone marrow-derived DCs were sensitized with fusion protein to obtain the fusion protein sensitized DC vaccines.B16F10 melanoma cells were transplanted into C57BL/6J male mice to construct a melanoma model (1×10^6 cells per mouse),and DC vaccine was injected for treatment. The antitumor efficacy of DC vaccine was explored by in vitro and in vivo experiments.Flow cytometry analysis showed that the fusion protein can effectively stimulate DC into differentiation and maturation;in the animal experiment,the inhibition rate of melanoma treated with G3G6-DC was 35.75%,that of HST1-DC group and combination group were 34.03% and 55.74%.It was initially proved that both G3G6-DC and HST1-DC can effectively inhibit the growth of transplanted tumors of melanoma B16F10 cells in mice,and the combination therapy is superior to the single therapy.