上调Foxp3基因表达抑制上皮卵巢癌细胞株SKOV3增殖的研究.doc

文档介绍：
上调 Foxp3 基因表达抑制上皮卵巢癌细胞
株 SKOV3 增殖的研究#
张海燕,孙红**
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(复旦大学附属妇产科医院妇科,上海 200011)
摘要:目的:构建 Foxp3 基因真核表达载体,探讨 Foxp3 基因对上皮性卵巢癌细胞株 SKOV3
增殖能力的影响及机制。方法:从人外周血 CD4+T 细胞中扩增、克隆 Foxp3 基因,连接到
真核表达载体 pEF-1α,构建表达质粒 pEF-1α-Foxp3;G418 压力筛选稳定表达 Foxp3 细胞
株(SKOV3/pEF-1α-Foxp3),以空载体转染细胞株为对照。K-8 法观察 Foxp3 基
因对 SKOV3 生物学行为的影响,Western blot 检测探索 Foxp3 相关的调控机制。结果:Foxp3
基因的真核表达载体成功构建;SKOV3/pEF-1α-Foxp3 的增殖速度显著低于 SKOV3/pEF-1
α细胞,并抑制 mTOR 信号通路激活。结论:上调 Foxp3 基因表达能显著抑制上皮性卵巢
癌细胞的增殖能力,为深入研究 Foxp3 基因在卵巢癌靶向治疗提供实验依据。
关键词:卵巢癌;Foxp3;真核表达载体;信号转导
中图分类号:
Effects of up-regulation Foxp3 expression on the
proliferation of epithelial ovarian cancer cells
ZHANG Haiyan, SUN Hong
(Department of Gynecology,The Obstetrics and Gynecology Hospital Affiliated Fudan University,
ShangHai 200011)
Abstract: Objective: To construct the eukaryotic expressing vector for Forkhead helix
transcription factor (Foxp3) and transfect it to human epithelial ovarian cancer cells for stable
expression of Foxp3 for investigating the role and mechanism of Foxp3 to inhibit tumor cells
activities. Methods: Eukaryotic expressing vector pEF-1α-Foxp3 was constructed and transfected
into SKOV3 cells by lipofectamine protocols. After G418 selection, the cells expressing Foxp3
stably(SKOV3/pEF1-α-Foxp3)were obtained. Multiple cellular and molecular approaches such
as cell growth assay, western blo