Ad.Ela对SKOV3细胞的转染效率及体外杀伤作用  

Adenovirus-mediated Ela gene therapy for SKOV3 cell line in vitro

作　　者：臧荣余[1] 秦慧莲[2] 陆浩均 张志毅[1] 蔡树模[1] 卢大儒[3] 

机构地区：[1]上海医科大学肿瘤医院妇科,200032 [2]上海医科大学免疫学实验室 [3]复旦大学遗传所

出　　处：《现代妇产科进展》2000年第1期13-15,共3页Progress in Obstetrics and Gynecology

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 AdRSVlacZ感染SKOV3细胞1次，基因转染效率732%。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.

关 键 词：卵巢肿瘤 腺病毒科 基因疗法 基因 

分 类 号：R737.310.5[医药卫生—肿瘤]