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

作　　者：井亮亮 苗梓韬 李曼曼[1] 叶佳[1] 金亮[1] 曹荣月[1] 龙军[2] 

机构地区：[1]中国药科大学生命科学与技术学院,南京210009 [2]南京中医药大学药学院,南京210029

出　　处：《中国药科大学学报》2017年第1期102-109,共8页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.81373232;No.31270985);国家基础科学人才培养基金资助项目(No.J1310032);江苏高校优势学科建设工程资助项目(PAPD)~~

摘　　要：探讨m GM-CSF/βh CG(GC)和h VEGF121/βh CG(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p ET-28a-m GM-CSF-X10-βh CGCTP37和p ET-28a-VEGF-M2-X10-βh CG-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h VEGF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。This study aimed at investigating the inhibitory effects and the anti-tumor mechanisms of co-administration of fusion proteins m GM-CSF/βh CG（ GC） and h VEGF121/βh CG（ VC） on RM-1 prostatic cancer and B16F10 melanoma in the C57 BL/6J mouse model.Two recombinant stains containing p ET-28a-m GM-CSF-X10-βh CGCTP37 and p ET-28a-VEGF-M2-X10-βh CG-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 C57 BL/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-h VEGF 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.

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 

分 类 号：R73-36[医药卫生—肿瘤]