肿瘤坏死因子-α基因联合氟尿嘧啶治疗胰腺癌探究.docVIP

肿瘤坏死因子-α基因联合氟尿嘧啶治疗胰腺癌探究 　　　【摘要】　目的　探讨肿瘤坏死因子(TNF)-α基因与氟尿嘧啶(5-FU)联合治疗胰腺癌的作用。方法　应用MoMLV逆转录病毒载体，将TNF-α基因转导致人胰腺癌细胞株PC-3，经氨基糖甙类新霉素衍生物Geneticin(G418)筛选获得了抗性克隆。用逆转录聚合酶链方法检测胰腺癌细胞株转染TNF-α的基因表达情况，同时用放线菌素D处理后的小鼠成纤维肉瘤细胞L929检测转基因细胞表达TNF-α的活性，并观察TNF-α基因和5-FU联合治疗胰腺癌的协同作用。结果　TNF-α基因在人胰腺癌中整合并表达有活性的TNF-α。转染TNF-α基因的细胞生长受抑制。TNF-α基因与5-FU联合应用提高了对胰腺癌的抑制作用。结论　TNF-α基因与5-FU联合应用对胰腺癌增殖的抑制有协同作用，并可减少5-FU的药量，对胰腺癌的综合治疗有潜在的临床应用价值。 　　Combination therapy of pancreatic cancer with tumor necrosis factor alpha gene and5-Fluorouracil 　　【Abstract】 Objective to investigate the synergistic antitumor effects of tumor necrosis factor alpha(TNF-α) gene and 5-Fluorouracil (5-FU) therapy in pancreatic cancer.Methods retroviral vector MoMLV was used to introduce human TNF-a gene into human pancreatic carcinoma cell line PC-3. The G418-resistant colonies were isolated and cloned. Expression of TNF-a gene was evaluated by RT-PCR. The activity of tNFa expressed by transducted pancreatic cancer cell strain was measured by rat fibrosarcoma cell L929 which was processed by Actinomycin D. Finally the synergistic antitumor effects of TNF-a gene and 5-FU were observed.Results rT-PCR analysis indicated t hat the integration and expression of TNF-a gene was seen only in the transducted cells. Using a bioassay method, different gene transducted PC-3 cells can secrete different amount of TNF-a.Growth of the cells transfected with TNF-α gene was inhibited. A combination of TNF-α and 5-FU could increase the inhibitory effects on pancreatic carcinoma.Conclusion combination therapy with TNF-a gene and 5-FU has synergism to inhibit pancreatic cancer cell proliferation, which can decrease the dose of 5-FU, and has potential value to clinical treatment of pancreatic cancer. 　　【Key words】 Human tumor necrosis factor; gene moddification; Human pancreatic carcinoma; 5-Fluorouracil 　　我们采用逆转录病毒介导肿瘤坏死因子(TNF)-α基因修饰人胰腺癌细胞后联用氟尿嘧啶(5-FU)，旨在为胰腺癌基因治疗提供新的应用策略。材料和方法 　　1.基因转染：逆转录病毒载体人工包装TNF-α质粒(PLhTNF-α sN)含有内切酶EcoRI和BamHI位点。外源性基因猴病毒40(SV40)为启动