OSKM诱导过表达c-Jun肝癌细胞重编程
Acta Anatomica Sinica(2017)
Abstract
目的 构建OSKM诱导的过表达c-Jun肝癌细胞(C3A-c-Jun)重编程细胞系,探讨外源性c-Jun对肝癌细胞重编程的影响.方法 通过C3A细胞稳定过表达c-Jun后进行OSKM诱导重编程,通过碱性磷酸酶染色,Real-time PCR,免疫印迹法,免疫荧光等技术对细胞进行鉴定,建立C3A-c-Jun诱导肿瘤干细胞系C3A-c-Jun-iCSCs.结果 C3A-c-Jun-iCSCs克隆呈圆顶状,边缘圆钝,克隆内细胞小且排列紧密,多层分布,碱性磷酸酶染色阳性,mRNA和蛋白水平均可表达多潜能性标记物OCT4、SOX2;C3A-c-Jun-iCSCs组外源性和内源性Sox2的表达量均明显高于C3A-iCSCs对照组;在重编程过程中c-Jun持续表达.结论 OSKM诱导C3A、C3A-c-Jun肝癌细胞重编程,分别建立C3A-iCSCs和C3A-c-Jun-iCSCs细胞系,发现外源性c-Jun通过上调Sox2基因的表达,启动下游一系列反应,对肝癌细胞重编程过程起促进作用.
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Key words
c-Jun,Induce,Cancer stem cell,Reprogramming,Western blotting,C3A cell
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