将融合蛋白hVEGF121-M2-X10-βhCG-CTP37作为免疫原,并分别联合化学药物环磷酰胺(CTX)和多西他赛(DTX),探索其对C57BL/6J小鼠B16F10黑色素瘤的抑制效应。将h VEGF121/βhCG融合蛋白(VC)分别与CTX和DTX联合给药(VCC、VCD),免疫接种B16F10黑色素瘤小鼠,对免疫小鼠皮内肿瘤组织周围毛细血管进行计数、测量各组小鼠瘤块体积和重量、对肿瘤组织进行切片分析、检测免疫小鼠脏器指数、MTT法检测脾细胞增殖、LDH法检测特异性杀伤活性。皮内肿瘤组织周围血管计数结果显示,与NS组比,DTX组、VC组以及VCD组均能高效抑制黑色素瘤血管生成(P〈0.01);皮下肿瘤抑制结果显示,与NS组相比,DTX组和VCD组抑瘤效果最为显著,抑瘤率分别为60.0%,70.0%,其中VCD组抗肿瘤效果优于VC组或DTX组给药组(P〈0.01);肿瘤组织病理学切片结果显示,VCD组的病变程度和炎细胞浸润程度最显著;对小鼠脾脏指数分析结果显示,与NS组相比,DTX组和VCD组的脾脏指数均显著增高(P〈0.05);抗肿瘤免疫实验中:与NS组相比,VCD组提高了脾细胞增殖能力和细胞毒作用(P〈0.05),且VCD组小鼠的脾细胞杀伤率在效靶比为20∶1、40∶1和80∶1时均具有显著性差异(P〈0.05)。h VEGF121/βhCG融合蛋白与化学药物DTX初步表现出协同抗肿瘤的效果,而与化学药物CTX未表现出协同抗肿瘤作用。
The fusion protein h VEGF121-M2-X10-βhCG-CTP37 was taken as an immunogen and combined with chemicals cyclophosphamide( CTX) and docetaxel( DTX) respectively to explore its effect of inhibition B16F10 melanoma on the C57 BL /6J mice. The fusion protein h VEGF121/ βhCG( VC) was combined with CTX and DTX( VCC,VCD),respectively. And then immunized with B16F10 melanoma of C57 BL /6J mice. Counting the capillaries around the tumor tissue within the immunized mice skin,the tumor volume and weight of each group were measured and then HE staining was used to analyze the slices of tumor tissues. Visceral index of each mouse was tested. MTT assay was carried out to detect proliferation of splenocyte. Cytotoxicity was detected by LDH. The results of counting vessels surrounding of the tumor tissue showed that compared with normal saline( NS),DTX group,VC group and VCD group can inhibit melanoma angiogenesis effectively( P〈 0. 01). The results of inhibition to subcutaneous tumor showed that compared with NS group,DTX group and VCD group anti-tumor effects are more remarkabe( P〈 0. 01),and the anti-tumor rates can get 60% and 70% respectively,besides that the anti-tumor effect of VCD group was more positive than the DTX or VC alone( P 〈0. 01). The histopathology of tumor sections showed that the degree of pathological changes and inflammatory cell infiltration of VCD group was the most significant. The spleen index analysis showed that compared with the NS group,the index in DTX group and VCD group was significanty higher( P 〈0. 05). Anti-tumor immunity assay showed that splenocyte proliferation and cytotoxic effects on tumor cells increased compared with NS group( P〈 0. 05),also,when the effector-target ratio in VCD group is 20 ∶ 1,40 ∶ 1 and80∶ 1,it showed significantly difference compared with group NS( P 〈0. 01). The combination between DTX and VC showed a synergetic anti-tumor effect while there was no synergetic anti-tumor effect between CTX and VC.