Ad Ela对SKOV3细胞的转染效率及体外杀伤作用

臧荣余　秦慧莲　陆浩均　张志毅　蔡树模　卢大儒

　　【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。
　　【关键词】　卵巢肿瘤；腺病毒科；基因疗法；基因，HER-2/neu
　　中图分类号　R737.31

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

Zang Rongyu,Qin Huilian,Lu Haojun
（Cancer Hospital of Fudan University School of Medicine,Shanghai，200032）

　　Abstract Objective:To test the feasibility and efficacy of adenovirus-mediated Ela gene therapy for ovarian cancer in vitro.Methods:Recombinant,replication-deficient adenoviral vectors were prepared by superinfection of 293 cells,and then purified.The efficacy of the adenovirus vector system to infect ovarian cancer cells was tested using different V/T ratios and different times(1-4) of Ad.RSVlacZ.SKOV3 cells(103 pre well)were infected once with 2×104adenovirus.The cells were harvested and counted on different days for 7 days to generate the in vitro growth curve.Expression of HER-2/neu P185 protein was detected using FACS.Results:The transduction efficacy 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 ovarian 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.
　　Key words　Ovarian neoplasms;Adenoviidae;Gene therapy;Gene,HER-2/neu

　　目前，基因治疗所面临的诸

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