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目的研究腺病毒介导的组织途径抑制因子2(tissuefactorpathwayinhibitor-2,TFPI-2)基因对喉鳞状细胞癌(简称鳞癌)生长的抑制作用。方法扩增并鉴定携带人-TFPI-2的重组腺病毒(adenovirusesTFPI-2,AdTFPI-2)。建立喉鳞癌荷瘤裸鼠动物模型,治疗组9只裸鼠瘤周注射AdTFPI2的重组腺病毒2×1011空斑形成单位(plaqueformingunits,PFU)共3次,对照组9只裸鼠瘤周注射等量的空载体,观察治疗结束后1周两组肿瘤质量和体积的差异。运用透射电镜观察肿瘤形态学变化;运用免疫组化方法检测肿瘤组织中增殖核抗原的表达。结果扩增后的AdTFPI-2的病毒滴度为2.8×1012PFU/ml。AdTFPI2治疗组裸鼠肿瘤平均体积和质量分别为(1.20±0.34)cm3和(1.52±0.39)g,明显小于空载体对照组的(2.08±0.52)cm3和(2.67±0.47)g(P<0.01)。透射电镜观察到AdTFPI2治疗组肿瘤呈凋亡改变,免疫组化结果显示AdTFPI-2治疗组的增殖细胞核抗原指数(54.9%±12.4%)明显低于对照组(75.8%±11.2%,P<0.01)。结论瘤周注射携带人TFPI2的重组腺病毒可抑制裸鼠体内喉鳞癌的生长。
Objective To study the inhibitory effect of adenovirus-mediated TFPI-2 gene on the growth of laryngeal squamous cell carcinoma (SCC). Methods Recombinant adenovirus carrying human TFPI-2 (adenoviruses TFPI-2, AdTFPI-2) was amplified and identified. The animal model of laryngeal squamous cell carcinoma in nude mice was established. In the treatment group, 9 nude mice in the treatment group were injected with 2 × 1011 recombinant plaqueforming units (PFU) The same amount of empty vector to observe the difference between the two groups of tumor mass and volume after 1 week of treatment. The morphological changes of tumor were observed by transmission electron microscope. The expression of proliferating nuclear antigen in tumor tissue was detected by immunohistochemistry. Results The amplified virus titer of AdTFPI-2 was 2.8 × 10 12 PFU / ml. The average volume and mass of tumors in nude mice treated with AdTFPI2 were (1.20 ± 0.34) cm3 and (1.52 ± 0.39) g, respectively, which were significantly lower than those in the control group (2.08 ± 0.52 cm3 and 2.67 ± 0.47g, P <0.01 ). The results of transmission electron microscopy revealed that the AdTFPI2-treated tumors showed apoptotic changes, and the immunohistochemistry results showed that the Ad-NPT-2 treated group had significantly lower proliferative cell nuclear antigen index (54.9% ± 12.4%) than that of the control group (75.8% ± 11.2%, P <0.01) ). Conclusion Peritumor injection of recombinant adenovirus carrying human TFPI2 can inhibit the growth of laryngeal squamous cell carcinoma in nude mice.