OSKM诱导过表达c-Jun肝癌细胞重编程
发布时间：2018-10-05 12:04
【摘要】：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-i CSCs。结果 C3A-c-Jun-i CSCs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-i CSCs组外源性和内源性Sox2的表达量均明显高于C3A-i CSCs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-i CSCs和C3A-c-Jun-i CSCs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。
[Abstract]:Objective to construct OSKM induced reprogramming cell line of c-Jun cells (C3A-c-Jun) and to investigate the effect of exogenous c-Jun on reprogramming of hepatoma cells. Methods after overexpression of c-Jun in C3A cells, OSKM induction reprogramming was carried out, and C3A-c-Jun induced tumor stem cell line C3A-c-Jun-i CSCs. was established by using alkaline phosphatase staining and real-time PCR, immunoblotting, immunofluorescence and other techniques. Results the C3A-c-Jun-i CSCs clones were domed and obtuse at the edge. The cells in the clones were small and tightly arranged and distributed in multiple layers. The expression of exogenous and endogenous Sox2 in OCT4,SOX2;C3A-c-Jun-i CSCs group was significantly higher than that in C3A-i CSCs control group, and the expression of c-Jun continued during reprogramming. Conclusion OSKM induces reprogramming of C3A- C3A-c-Jun hepatoma cells and establishes C3A-i CSCs and C3A-c-Jun-i CSCs cell lines respectively. It is found that exogenous c-Jun promotes the reprogramming process of HCC cells by up-regulating the expression of Sox2 gene and initiating a series of downstream reactions.
【作者单位】： 北京大学医学部细胞生物学系;北京东方亚美基因科技研究院有限公司;
【基金】：国家自然科学基金(81572313) 广东省省级科技计划项目(2015B020229002)
【分类号】：R735.7
[Abstract]:Objective to construct OSKM induced reprogramming cell line of c-Jun cells (C3A-c-Jun) and to investigate the effect of exogenous c-Jun on reprogramming of hepatoma cells. Methods after overexpression of c-Jun in C3A cells, OSKM induction reprogramming was carried out, and C3A-c-Jun induced tumor stem cell line C3A-c-Jun-i CSCs. was established by using alkaline phosphatase staining and real-time PCR, immunoblotting, immunofluorescence and other techniques. Results the C3A-c-Jun-i CSCs clones were domed and obtuse at the edge. The cells in the clones were small and tightly arranged and distributed in multiple layers. The expression of exogenous and endogenous Sox2 in OCT4,SOX2;C3A-c-Jun-i CSCs group was significantly higher than that in C3A-i CSCs control group, and the expression of c-Jun continued during reprogramming. Conclusion OSKM induces reprogramming of C3A- C3A-c-Jun hepatoma cells and establishes C3A-i CSCs and C3A-c-Jun-i CSCs cell lines respectively. It is found that exogenous c-Jun promotes the reprogramming process of HCC cells by up-regulating the expression of Sox2 gene and initiating a series of downstream reactions.
【作者单位】： 北京大学医学部细胞生物学系;北京东方亚美基因科技研究院有限公司;
【基金】：国家自然科学基金(81572313) 广东省省级科技计划项目(2015B020229002)
【分类号】：R735.7
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