胞嘧啶脱氨酸基因治疗人胰腺癌的实验研究

探讨腺病毒介导的大肠杆菌胞嘧啶脱氨酶基因用于人胰腺癌基因疗法的可行性。方法将含癌胚抗原启动子的重组腺病毒感染人胰腺癌ＳＷ１９９０细胞和Ｃａｐａｎ－２细胞以及人宫颈癌Ｈｅｌａ细胞,用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）和Ｗｅｓｔｅｒｎｂｌｏｔ检测ＣＤ基因在细胞中的表达,以ＭＴＴ法比较细胞对５－氟胞嘧（５－ＦＣ）的敏感性差异。建立胰腺癌裸鼠皮下移植瘤模型,观察ＣＤ基因的原位治疗效应及安全性。结果腺病毒介

Experimental study on human pancreatic carcinoma treated by cytosine deaminase gene therapy

Objective To explore the possibility of Escherichia coli cytosine deaminase (CD) gene on human pancreatic carcinoma gene therapy. Methods Recombinant adenoviruses containing a carcinoembryonic antigen (CEA) promoter were transiently introduced into SW1990, Capan 2 and Hela cells, separately. The expression of CD gene mRNA was examined by RT PCR. CD protein level in the transduced cells was analyzed by Western blotting. The sensitivity of the cells to 5 fluorocytosine (5 FC) was determined by MTT assay. Results A specific expression of cytosine deaminase gene by adenovirus mediated transfer exhibited only in SW1990 cells (CEA producing). Transduction of CD gene resulted in significant sensitivity of SW1990 cells to 5 FC. The anticancer effect was seen in vivo in SW1990 xenografts nude mice with in situ CD gene transduction. Conclusion The targeted expression of CD gene combined prodrug 5 FC may be a potential approach for gene therapy for human pancreatic carcinoma.