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In vitro experimental study of gene therapy for ovarian cancer with thymidine kinase gene of herpes simplex virus mediated by a non-viral GE7 delivery system
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF OBSTETRICS AND GYNECOLOGY
- Issue:
- 10
- DOI:
- 10.3760/j.issn:0529-567X.2003.10.009
- Key Word:
- 卵巢肿瘤;基因疗法;疱疹病毒1型,人;胸苷激酶
Abstract： 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10 mg/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.
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