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目的 :观察经动脉插管导入肿瘤坏死因子α(TNF -α)和白介素2(IL -2)重组腺病毒对转移性大鼠肝癌模型的治疗作用。材料和方法 :用293细胞扩增人TNF -α和IL -2重组腺病毒 ,并测定病毒滴度 ;9组大鼠于胃十二指肠动脉分别注入Ad.lacZ(2×109pfu/ml) ,采用X -Gal染色方法检测lacZ基因在各个脏器的表达 ;Walker256乳癌细胞感染人TNF重组腺病毒后 ,观察其体外增殖能力、皮下致瘤性的变化及进行肿瘤局部病理学检查 ;瘤体局部注射TNF重组腺病毒 ,观察荷瘤大鼠的肿瘤生长情况及生存期 ;取大鼠乳腺癌细胞株(Walker256)转染至肝脏的荷瘤大鼠动物模型 ,分为hTNF组 ,人IL -2(hIL -2)组 ,hTNF +hIL -2组 ,病毒对照组(lacZ组)和培养液对照组进行治疗 ,观察大鼠生存期。结果 :所制备的人TNF -α和IL -2重组腺病毒滴度为2×109pfu/ml和2.1×109pfu/ml;实验组于12h取材 ,经染色lacZ基因在肝脏中即有表达 ,而此时脾、肺、肾等内脏器官尚未见表达 ,21d该基因在肝脏仍有表达 ,而在其他脏器如脾、肺及肾表达仅维持14d ;转染基因瘤株体外增殖能力和皮下致瘤性显著下降 ,瘤体周围有大量淋巴细胞及单核细胞浸润 ;肿瘤局部注射Ad.hTNF能显著抑制Walker256的生长 ,并延长荷瘤大鼠的生存期 ;单独注射TNF -α或IL -2重组腺病毒能有效
Objective : To observe the therapeutic effect of recombinant adenovirus containing tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2) introduced through transarterial cannula in a metastatic rat hepatoma model. Materials and Methods: Human TNF-α and IL-2 recombinant adenoviruses were amplified with 293 cells and virus titers were measured. Adjuvant rats were injected with Ad.lacZ (2×109 pfu/ml) in 9 groups. X-Gal staining was used to detect the expression of lacZ gene in various organs. After Walker256 breast cancer cells were infected with human TNF recombinant adenovirus, the proliferation ability, subcutaneous tumorigenicity and tumor local pathology were observed. Local injection of TNF recombinant adenovirus was used to observe the tumor growth and survival of tumor-bearing rats. The rat model of tumor-bearing rat transfected with liver (Walker 256) into liver was divided into hTNF group and human IL- 2 (hIL -2) group, hTNF + hIL -2 group, virus control group (lacZ group) and culture fluid control group were treated to observe the survival period of rats. Results: The titer of recombinant human adenovirus TNF-α and IL-2 was 2×109 pfu/ml and 2.1×109 pfu/ml. The experimental group was obtained at 12 h and the stained lacZ gene was expressed in the liver. No expression was found in the internal organs of the spleen, lungs, kidneys, etc. The gene was still expressed in the liver at 21 days, but only maintained for 14 days in other organs such as spleen, lung, and kidney; the in vitro proliferation ability and subcutaneous tumorigenicity of the transfected tumor strains were observed. Significantly decreased sex, there are a large number of lymphocytes and mononuclear cell infiltration around the tumor; local injection of Ad.hTNF can significantly inhibit the growth of Walker256, and prolong the survival of tumor-bearing rats; injection of TNF-α or IL-2 alone Adenovirus is effective