摘要
目的 探讨靶向非病毒载体GE7在卵巢癌细胞株CAOV3细胞中的转导效率 ,及其介导的Ⅰ型单纯疱疹病毒胸腺嘧啶核苷激酶 (HSV1 tk) /更昔洛韦 (GCV)基因治疗系统在卵巢癌治疗中的应用。方法 构建由表皮生长因子受体 (EGFR)介导的非病毒载体GE7,分别与外源基因 [即报告基因 β 半乳糖苷酶 (β gal)和治疗基因HSV1 tk]构成载体复合物 ,体外转导CAOV3细胞 ,通过 5 溴 4 氯 3 吲哚 β D半乳糖苷 (X gal)染色、核酸分子印记杂交分析 ,观察非病毒载体GE7对外源基因 β gal及HSV1 tk基因的转导情况 ;将GCV加入转导HSV1 tk基因的CAOV3细胞 ,通过细胞生长抑制曲线、流式细胞仪分析等 ,观察其对肿瘤细胞的杀伤作用。结果 β gal基因的转导效率可达 80 % ,呈瞬时表达 ;加入 10mg/L的GCV时 ,转导HSV1 tk基因的CAOV3细胞的生长抑制率可达 95 % ;流式细胞仪分析显示 ,CAOV3细胞S期比例上升 ,最高达 90 % ,凋亡指数高达 30。结论 GE7载体系统能高效地将外源基因导入CAOV3细胞 ,GE7/HSV1 tk /GCV基因治疗系统在卵巢癌治疗中可能具有良好的应用前景。
Objective To investigate gene transfer efficiency of a novel target non-viral vector GE7 and effects of herpes simplex virus thymidine kinase (HSV 1-tk)/ganciclovir(GCV) mediated by it in vitro. Methods The epidermal growth factor receptor (EGF-R) target gene delivery system GE7 was constructed.Human ovarian cancer cell line CAOV3 was transfected in vitro with β-galactosidase(β-gal) as reporter gene and HSV 1-tk gene as therapeutic gene using this gene delivery system.By means of the assay of X-gal staining, Northern blotting, cell growth-inhibiting curve and so on,the transferring efficiency of exogenous genes and killing effects are observed. Results It showed that gene transfer efficiency is over 80%.When 10 mg/L GCV was put into ovarian cells transfected with HSV 1-tk gene, 95% of cells were killed, and the apoptosis ratio reached up to 30. Conclusions The GE7 gene delivery system is an effective and safe delivery system.GE7/ HSV 1-tk /GCV therapeutic gene system is appraising for ovarian cancer.
出处
《中华妇产科杂志》
CAS
CSCD
北大核心
2003年第10期621-624,共4页
Chinese Journal of Obstetrics and Gynecology
基金
国家自然科学基金资助项目 (3 980 0 14 4)
