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
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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。
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关键词:
- 血管内皮生长因子(VEGF) /
- 人绒毛膜促性腺激素(hCG) /
- 鼠源粒细胞巨噬细胞集落刺激因子(mGM-CSF) /
- 前列腺癌 /
- 黑色素瘤
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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