VEGF融合蛋白疫苗联合环磷酰胺抗小鼠H22肝癌实验研究

Improved antitumor efficacy by combination treatment with recombined VEGF protein vaccine and cyclophosphamide in H22 hepatocellular carcinoma bearing-mice

目的探讨分子佐剂修饰的VEGF融合蛋白疫苗与低剂量环磷酰胺(CTX)能否产生协同抗肝癌作用。方法建立小鼠H22肝癌模型,将BALB/c小鼠随机分为4组:生理盐水对照组、CTX单药组、VEGF蛋白疫苗单药组(V2组)、V2与CTX联合治疗组(V2+CTX)。分别在H22肝癌皮下移植瘤及皮内肿瘤模型中评价联合治疗方案抗肿瘤生长及抗血管生成的能力,并通过Western blot及ELISA方法检测抗-VEGF抗体水平。结果皮下移植瘤模型结果显示V2+CTX联合治疗组的肿瘤体积、平均瘤重低于各单药治疗组(P<0.05 vs V2,P<0.01 vs CTX);在皮内肿瘤血管模型中,联合治疗对肿瘤新生血管的抑制作用最为明显,与单独的V2及CTX组相比,差异均有统计学意义(P<0.05 vs V2,P<0.01 vs CTX);Western blot及ELISA的结果显示,V2+CTX联合治疗诱导小鼠产生了高水平的特异性抗-VEGF抗体。结论低剂量CTX与重组VEGF融合蛋白疫苗联合治疗有协同抗肝癌作用。

Aim To investigate the antitumor and antiangiogenic effects of combined low-dose cyclophosphamide（CTX）and recombined VEGF protein vaccine.Methods In this experiment, H22 hepatocellular carcinoma model was established in BALB/c mice.Mice were randomly divided into four groups： control group, CTX group（CTX）, VEGF protein vaccine group（V2）and CTX plus V2 group（CTX＋V2）.The anti-tumor efficacy and antiangiogenic effect were investigated using a subcutaneous tumor model and an intradermal tumor model.Western blot and ELISAwere further adopted to detect the specific anti-VEGF antibody.Results CTX＋V2 group displayed a lower tumor volume and tumor weight than either the single therapy group in the subcutaneous tumor model（P〈0.05 vs V2,P〈0.01 vs CTX）.Meanwhile, CTX＋V2 was more effective for antagonizing tumor-associated angiogenesis compared with either the single therapy（P〈0.05 vs V2, P〈0.01 vs CTX）.After CTX＋V2 immunization, high titer of anti-VEGF antibody was detected by ELISA and verified by Western blot.Conclusion The therapy of CTX combined with V2 has significant synergistic effect against H22 hepatocellular carcinoma.