腺病毒介导的组织途径抑制因子2基因抑制喉鳞状细胞癌生长的实验研究

Experimental investigation of effects of adenovirus transducted tissue factor pathway inhibitor-2 gene on growth of laryngeal squamous carcinoma

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 investigate the inhibitory effects of adenovirus transducted TFPI-2 gene on the growth of laryngeal squamous carcinoma. Methods Recombinant adenoviruses carrying human (TFPI-2) gene were amplified and identified. The nude mouse model of laryngeal squamous carcinoma was established by intracutaneous injection of Hep-2 cells. Mice in the treated group were injected with recombinant adenoviruses with Ad-TFPI-2 (adenoviruses-TFPI-2) in peritumor tissue while mice in control group were injected with equivalent null plasmids. After treatment, the tumor weight and volume of tumor in each mouse were measured respectively. The morphological changes of tumor cells were observed using transmission electron microscope and proliferating cell nuclear antigen (PCNA) expression was examined using immunohistochemistry. Results The Ad-TFPI-2 virus titer was 2.8×10^(12) PFU/ml after amplification. The average tumor weight and volume in Ad-TFPI-2 treated group were (1.20±0.34)cm^3 and the volume ((1.52±)0.39)g, which were significantly lower than the tumor weight (2.08±0.52)cm^3 and (2.67±0.47)g in the control group(P<0.01). Apoptosis was observed in the tumors of Ad-TFPI-2 treated group. The PCNA index in Ad-TFPI-2 group was (54.9%±12.4%),which was obviously lower than that (75.8%±11.2%)in control group(P<0.01). Conclusion Peritumor injection of Ad-TFPI-2 can inhibit the growth of laryngeal squamous carcinoma in nude mouse model.