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融合蛋白hVEGF121/βhCG与mGM-CSF/βhCG联合抗肿瘤作用及其机制

Anti-tumor effect and its mechanism of co-administration of fusion proteins hVEGF121/βhCG and mGM-CSF/βhCG

  • 摘要: 探讨mGM-CSF/βhCG(GC)和hVEGF121/βhCG(VC)两种融合蛋白联用对C57BL/6J小鼠RM-1前列腺癌和B16F10黑色素瘤的抑制效果,并对其抗肿瘤作用机制进行初步研究。采用含有pET-28a-mGM-CSF-X10-βhCGCTP37和pET-28a-VEGF-M2-X10-βhCG-CTP37的两个重组菌进行乳糖诱导表达,分离纯化融合蛋白。将两种蛋白制备抗肿瘤蛋白疫苗(VC蛋白疫苗和GC蛋白疫苗),并将两蛋白混合制备联合蛋白疫苗(VGC蛋白疫苗),免疫C57BL/6J接瘤小鼠,测量瘤块生长状态;检测免疫小鼠脾细胞增殖以及对肿瘤细胞的细胞毒作用;ELISA检测小鼠体内细胞因子IFN-γ和抗hVEGF抗体浓度。结果发现,在前列腺癌和黑色素瘤小鼠移植瘤模型中,与生理盐水NS组相比,各实验组均具有统计学意义(P<;0.05)。其中VGC组抗肿瘤效果优于GC组和VC组(P<;0.01),且对前列腺癌和黑色素瘤的抑瘤率分别达到(41.7±0.83)%和(46.4±1.27)%。由此可见,VC和GC两种蛋白联合后,通过发挥抗肿瘤免疫和抗肿瘤血管生成双重作用,高效地抑制了RM-1前列腺癌和B16F10黑色素瘤的生长。

     

    Abstract: This study aimed at investigating the inhibitory effects and the anti-tumor mechanisms of co-administration of fusion proteins mGM-CSF/βhCG(GC)and hVEGF121/βhCG(VC)on RM-1 prostatic cancer and B16F10 melanoma in the C57BL/6J mouse model. Two recombinant stains containing pET-28a-mGM-CSF-X10-βhCGCTP37 and pET-28a-VEGF-M2-X10-βhCG-CTP37 were induced by lactose to express fusion proteins. The fusion proteins were separated and purified to prepare the anti-tumor protein vaccines(VC protein vaccine and GC protein vaccine), which were then mixed to prepare a combined protein vaccine named VGC protein vaccine. The prostatic cancer and melanoma tumor-bearing mice C57BL/6J were immunized with described vaccines, then the growth of each tumor was measured; splenocyte proliferation of immunized mice was detected; and the cytotoxic effects of the vaccine on tumor cells were tested. After that, the in vivo concentrations of IFN-γ and anti-hVEGF antibodies were investigated by ELISA. The difference between each experimental group and normal saline group(NS)was statistically significant in both tumor-bearing mouse models(P< 0. 05)respectively. Besides, VGC group exhibited significantly better anti-tumor effect compared with the GC and VC groups with the anti-tumor rate(41. 7±0. 83)% and(46. 4±1. 27)% for prostatic cancer and melanoma tumor, respectively. The co-administration of the two proteins, VC and GC, could inhibit the growth of RM-1 prostatic tumor and B16F10 melanoma effectively via anti-tumor immunity and anti-tumor angiogenesis.

     

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