摘要
目的：研究腺病毒Ela质粒对HER-2/neu高表达卵巢癌细胞系SKOV3的转染效率和体外治疗作用。方法：脂质体介导腺病毒载体重组，腺病毒扩增、纯化、滴度测定，不同滴度病毒转染卵巢癌细胞的效率分析，以20：1的Ad.Ela抑制SKOV3细胞生长。结果：体外20：1 AdRSVlacZ感染SKOV3细胞1次，基因转染效率732%。此后增加病毒量，转染效率曲线接近平台。103*"SKOV3细胞分别以2×104 PFU的Ad.Ela+、Ad.Ela-和PBS感染1次，结果显示，Ad.Ela+治疗后卵巢癌细胞生长明显抑制，FACS检测转染后的SKOV3细胞，P185蛋白的表达明显下降。结论：Ad.Ela对HER-2/neu高表达细胞SKOV3的生长有显著抑制作用。
\ Objective:To test the feasibility and efficacy of adeno virus-mediated Ela gene therapy for ovarian cancer in vitro.Methods:Recombinant,replication-deficient adenoviral vectors were prepared by superi nfection of 293 cells,and then purified.The efficacy of the adenovirus vector sy stem to infect ovarian cancer cells was tested using different V/T ratios and di fferent times(1-4) of Ad.RSVlacZ.SKOV3 cells(103 pre well)were infected once w ith 2×104adenovirus.The cells were harvested and counted on different days fo r 7 days to generate the in vitro growth curve.Expression of HER-2/neu P185 prot ein was detected using FACS.Results:The transduction effica cy increased with higher V/T ratios and reached a plateau at the 20:1 ratio.When Ad.Ela(+)was used to transduce the HER-2/neu overexpressing human ov arian cancer cell line SKOV3,tumor cell growth in vitro was greatly inhibited by Ela transducion.HER-2/neu P185 protein synthesis tested by FACS was significantly lower in the SKOV3 cells transduced by Ad.Ela(+) than that in the control groups.Conclusions:In this system,adenovirus can efficiently deliver a functional tumor suppressor gene into tumor cells,and the growth of SKOV3 cell was greatly inhibited by Ad.Ela.
出处
《现代妇产科进展》
CSCD
2000年第1期13-15,共3页
Progress in Obstetrics and Gynecology
关键词
卵巢肿瘤
腺病毒科
基因疗法
基因
Ovarian neoplasms
Adenoviidae
Gene therapy
Gene,HER-2 /neu
