摘要
目的比较靶向非病毒载体GE7系统介导Ⅰ型单纯疱疹病毒胸腺嘧啶核苷激酶(HSV1-tk)基因一次及持续转导卵巢癌细胞的转导效率及杀伤效应。方法靶向非病毒载体系统GE7分别与标志基因β-半乳糖苷酶(β-gal)基因和治疗基因HSV1-tk基因构成载体复合物,一次及持续体外转导卵巢癌细胞CaOV3,通过5-溴-4-氯-3-吲哚-β-D-半乳糖吡喃糖甙(X-gaI)染色比较GE7系统一次及持续转导外源基因的转导效率。将丙氧鸟苷(GCV)加入一次及持续转导HSV1-tk基因的卵巢癌细胞,通过细胞生长抑制曲线、流式细胞分析等,观察GCV对一次及持续转导GE7/HSV1-tk的卵巢癌细胞的杀伤作用。结果X-gal染色显示,GE7-次转导外源基因的平均转导效率可达80%,持续转导达85%。当GCV为1μg/mL时,GE7/HSV1-tk一次转导的生长抑制率达82%,凋亡指数为15,而持续转导可达90%,凋亡指数为30。在相同的GCV浓度下,持续转导GE7/pCMV-tk卵巢癌细胞生长抑制率显著高于一次转导(P<0.05)。结论GE7载体系统持续转导卵巢癌细胞较一次转导的平均转导效率高,持续转导的GE7/HSV1-tk/GCV基因治疗系统杀伤作用更大。
Objective To compare the transferring efficiency and killing effects of one time and continuous mediation with GE7, a non-viral targeted delivery system, in transfection of thymidine kinase gene of herpes simplex virus into ovarian cancer cells. Methods GE7 was used to prepare recombinants with β-gal and HSV1-tk; the recombinants were then used to transfect human ovarian cancer line CaOV3 once and continuously. β-galactosidase staining was used to compare the efficiencies of one time and continuous mediation with GE7 system. GCV was introduced into HSV1-tk transfected ovarian cells. Through drawing the cell growth curve and flow cytometory, the killing effect of GCV on once and continuous GE7/HSV1-tk transfected cells were observed. Results We found that the one time and continuous exogenous gene transfer efficiencies were about 80% and 85%, respectively. When 1 μg/mL ganciclovior was used to treat ovarian cell transfected with HSV1-TK gene, growth-inhibiting rates of ovarian cells of one time and continous transferring were 82% and 90%, respectively; their apoptosis indices were 15 and 30, respectively. Under same GCV concentration of GCV, continuous mediation of GE7/pCMV-tk transfection into ovarian cancer cells had more significant inhibitory effect than one time mediation （P 〈 0.05）. Conclusion Compared with one time mediation, continuous mediation of transfection with GE7 gene delivery system has higher efficiency. Continuous mediation of GE7/HSV1-tk/GCV therapeutic gene system has more powerful killing effect. （Shanghai Med J, 2007, 30：770-773）
出处
《上海医学》
CAS
CSCD
北大核心
2007年第10期770-773,共4页
Shanghai Medical Journal
基金
国家自然科学基金面上资助项目(39800144)
