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高表达Twist基因促进人乳腺癌细胞MCF-7干细胞样细胞富集和迁移

, PP. 2083-2086

Keywords: Twist,乳腺细胞小球,人乳腺癌细胞MCF-

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Abstract:

目的探讨人乳腺癌细胞系MCF-7中高表达Twist基因对MCF-7干细胞特性细胞的影响。方法构建重组质粒pBABE-puro-myc-Twist,HEK293T细胞包装逆转录病毒,并感染MCF-7细胞,通过筛选获得成功转入Twist的MCF-7/Twist细胞和MCF-7/Vector对照细胞,再通过长程无血清悬浮培养方法富集具有干细胞特性的乳腺细胞小球(Mammosphere),并通过Westernblot法检测myc-Twist及Twist诱导后富集的Mammosphere相关CSC蛋白标记的表达,?Transwell?法检测细胞的迁移能力。结果成功构建逆转录病毒质粒pBABE-puro-myc-Twist;Twist基因在靶细胞内稳定表达;长程无血清悬浮培养富集出具有干细胞特性的Mammosphere;MCF-7/Twist细胞肿瘤干细胞样细胞富集能力增强(P<0.05);myc-Twist、Twist及SOX2、OCT4蛋白在MCF-7/TwistMammosphere细胞中表达上调;MCF-7/TwistMammosphere细胞迁移能力增加(P<0.05)。结论Twist促进MCF-7肿瘤干细胞样细胞富集能力增强,并促进其迁移。

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