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目的:观察口服携带HIV tip30与人IFN-γ基因减毒沙门菌(SL7207)基因治疗舌癌荷瘤裸鼠的疗效。方法:4周龄BALB/c雄性裸鼠25只,随机分为PBS对照组、单纯SL7207组、携带人IFN-γ基因SL7207-pCI-IFN治疗组、携带HIV tip30基因SL7207-pCI-tip30治疗组和同时携带HIV tip30与人IFN-γ基因SL7207-pCI-tip30/IFN联合治疗组5组。裸鼠下颌下区皮下接种人舌癌Tca8113细胞10 d后,口服相应治疗菌,每周1次,共3次。测定瘤体积及瘤重变化,计算抑瘤率,免疫组化检测肿瘤组织中IFN-γ、Tip30的表达,琼脂糖凝胶电泳检测细胞凋亡。采用SPSS15.0软件包对数据进行统计学分析。结果:肿瘤生长曲线、动物累积生存率及肿瘤抑瘤率指标均显示,PBS对照组肿瘤呈进行性生长;单纯SL7207组显示一定抑瘤效果(P<0.05);携带人IFN-γ基因SL7207-pCI-IFN治疗组和携带HIV tip30基因SL7207-pCI-tip30治疗组均具有较好疗效(P<0.05),但2组之间无显著差异;Tip30和IFN联合SL7207-pCI-tip30/IFN治疗组疗效最好(P<0.01)。表明Tip30和IFN联合具有协同抗肿瘤作用。免疫组化检测发现,口服携带不同目的基因重组菌的肿瘤组织内相应蛋白高表达。琼脂糖凝胶电泳分析发现,转tip30基因的肿瘤细胞基因组DNA表现出凋亡DNA“阶梯状”条带。结论:口服重组减毒沙门菌具有体内瘤体转基因的能力,而且可使同时携带的HIV tip30与人IFN-γ2个目的基因同时达到瘤体细胞内表达,发挥协同抗瘤作用,为舌癌的基因治疗提供了新的方法。
Objective: To observe the curative effect of oral administration of tip30 and human IFN-γ gene attenuated Salmonella typhimurium (SL7207) on tongue cancer-bearing nude mice. Methods: Twenty-five male BALB / c nude mice of 4 weeks old were randomly divided into PBS control group, simple SL7207 group, SL7207-pCI-IFN group carrying human IFN-γ gene and SL7207-pCI-tip30 group carrying HIV tip30 gene And group 5 with the combination of HIV tip30 and human IFN-γ gene SL7207-pCI-tip30 / IFN at the same time. The submandibular area of nude mice was subcutaneously inoculated with human tongue cancer Tca8113 cells for 10 d, and the corresponding therapeutic bacteria were orally administered once a week for 3 times. Tumor volume and tumor weight were measured to calculate the tumor inhibition rate. Immunohistochemistry was used to detect the expression of IFN-γ and Tip30 in tumor tissues. The apoptosis was detected by agarose gel electrophoresis. SPSS15.0 software package for statistical analysis of the data. Results: The tumor growth curve, the cumulative survival rate of animals and the tumor inhibition rate indicators showed that the tumors in the PBS control group progressed progressively. The SL7207 alone group showed some antitumor effects (P <0.05). The human IFN-γ gene SL7207- There was no significant difference between the two groups in the pCI-IFN treatment group and in the SL7207-pCI-tip30 treatment group carrying the tip30 gene of HIV (P <0.05) The best effect (P <0.01). Tip30 and IFN combination has a synergistic anti-tumor effect. The results of immunohistochemistry showed that the overexpression of the corresponding protein in the tumor tissue orally carrying the recombinant bacterium of different gene of interest. Agarose gel electrophoresis analysis showed that the genomic DNA of the tip30 transgenic tumor cells showed apoptotic DNA “staircase ” bands. Conclusion: Oral recombinant attenuated Salmonella has the ability of in vivo tumor gene transfection, and can simultaneously carry both HIV tip30 and human IFN-γ2 target genes in tumor cells at the same time to play a synergistic anti-tumor effect for tongue cancer Gene therapy offers a new way.