人内皮抑素基因治疗卵巢癌裸鼠移植瘤的研究

目的探索人内皮抑素基因治疗人肿瘤的可行性：方法将人卵巢癌(SKOV3)瘤组织或其细胞移植于裸鼠皮下，建立成裸鼠移植瘤模型；用高保真PCR从含有人内皮抑素基因的重组质粒中扩增出与人生长激素信号肽序列融合的人内皮抑素编码区，将此基因克隆入pGEM-T载体中，经序列测定证明其序列的正确性；将此基因克隆入真核表达载体pcDNA3。经免疫荧光试验证明能在CPS-1细胞中表达。结果用重组质粒注射已形成移植瘤的裸鼠，能显著抑制移植瘤的生长；将基因注射与肿瘤移植同时进行，基因注射不仅能部分抑制移植瘤的形成，而且能抑制移植瘤的生长。结论本研究构建的分泌型表达人内皮抑素的重组质粒可以用于肿瘤的基因治疗研究。

Study on Gene Therapy for Human Ovarian Carcinoma Using Nude Mouse Model by Direct Injection of Recombinant Plasmid Expressing Endostatin Gene

Objective To explore the feasibility of gene therapy for human tumors by direct injection of recombinant plasmids. Methods Mouse tumor model was established by transferring human ovarian carcinoma (SKOV 3) tissues or separated cells subcutaneously into the oxter of nude mice, with a successful rate of 100%. The coding region for human endostatin (hES) fused with the signal peptide sequence of human growth hormone was amplified by PCR from a commercial recombinant plasmid and the correctness of the hybrid gene was confirmed by sequence analysis following subcloning into pGEM-T vector. The hybrid gene was subcloned into a eukaryotic expression vector pcDNA3 and its expression in COS-1 cells was demonstrated by immunofluroscence. Results Significant inhibition of the tumor growth was observed in 6/6 mice after muscular injection of nude mice with established tumor with pcDNA-ES. When the injection was done before tumor transplantation, the tumor could not establish itself in five out of the eight mice, while established in all of the mice (4/4) without plasmid injection. Conclusion These data indicate that the {established} animal model and gene therapy strategy might be useful for further studies on treating human tumors.