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目的探讨携带凋亡素基因(vp3gene)的重组腺病毒载体(AdAFPvp3)治疗C57BL/6小鼠肝细胞癌模型的可行性。方法检测腺病毒AdAFPvp3、腺病毒AdCMV-eGFP滴度。建立c57BL/6小鼠皮下荷肝细胞癌模型(n=24),计算成瘤率。待小鼠皮下肿瘤长至5mm左右时,随机分为治疗组、AdCMV-eGFP对照组、PBS对照组,每组8只,分别于瘤内注射AdAFPvp3(5×108pfu/100μl)、AdCMV-eGFP(5×108pfu/100μl)和PBS(100μl),隔日1次,共2次。每日测量肿瘤体积并观察有无不良反应,注射后7d处死小鼠,计算抑瘤率。肿瘤、肝脏和脾脏均行病理检查,用TUNEL法鉴定vp3在体内诱导肝癌细胞死亡的方式。结果腺病毒AdAF-Pvp3、AdCMV-eGFP滴度均为5×109pfu/ml。皮下肝癌成瘤率100%。瘤内注射AdAFPvp3后,小鼠肿瘤体积较病毒AdCMV-eGFP对照组、PBS对照组明显缩小(P<0.05)。治疗组抑瘤率78.87%,明显高于AdCMV-eGFP对照组(5.01%,P<0.05),治疗未见明显不良反应。凋亡素在体内以凋亡方式诱导肝癌细胞死亡。结论重组腺病毒载体AdAFPvp3基因治疗能抑制小鼠实验性肝细胞癌生长。
Objective To investigate the feasibility of recombinant adenovirus vector (AdAFPvp3) carrying apoptin gene (vp3gene) in the treatment of hepatocellular carcinoma in C57BL/6 mice. Methods Adenovirus AdAFPvp3, adenovirus AdCMV-eGFP titers were detected. A subcutaneous hepatoma model of c57BL/6 mice was established (n=24) and the tumorigenesis rate was calculated. When mice subcutaneous tumors grow to about 5mm, they were randomly divided into treatment group, AdCMV-eGFP control group, and PBS control group, with 8 mice in each group. Intratumoral injection of AdAFPvp3 (5×10 8 pfu/100 μl) and AdCMV-eGFP were performed. 5 x 10<8> pfu/100 [mu]l) and PBS (100 [mu]l) every other day for a total of 2 times. The tumor volume was measured daily and any adverse reactions were observed. The mice were sacrificed 7 days after the injection and the inhibition rate was calculated. Tumors, livers, and spleens were all pathologically examined. The TUNEL method was used to identify the manner in which vp3 induced hepatoma cell death in vivo. Results The titer of adenovirus AdAF-Pvp3 and AdCMV-eGFP was 5×10 9 pfu/ml. The tumor formation rate of subcutaneous hepatocellular carcinoma is 100%. After intratumoral injection of AdAFPvp3, the tumor volume of the mice was significantly smaller than that of the virus AdCMV-eGFP control group and PBS control group (P<0.05). The inhibitory rate of the treatment group was 78.87%, which was significantly higher than that of the AdCMV-eGFP control group (5.01%, P<0.05). There was no obvious adverse reaction after treatment. Apoptin induces death of hepatoma cells by apoptosis in vivo. Conclusion The recombinant adenoviral vector AdAFPvp3 gene therapy can inhibit the growth of experimental hepatocellular carcinoma in mice.