解剖学报

解剖学报 ›› 2017, Vol. 48 ›› Issue (3): 296-302.doi: 10.16098/j.issn.0529-1356.2017.03.008

• 肿瘤生物学 • 上一篇    下一篇

OSKM诱导过表达c-Jun肝癌细胞重编程

苏铭1 韩烁1 张明智1 李睿智1 魏示若1 曾沃坦2 刘进稳2 沈丽1*   

  1. 1. 北京大学医学部细胞生物学系，北京 100191; 2. 北京东方亚美基因科技研究院有限公司，北京 100078
  • 收稿日期:2016-11-07 修回日期:2017-01-03 出版日期:2017-06-06 发布日期:2017-09-19
  • 通讯作者: 沈丽 E-mail:shenli@bjmu.edu.cn
  • 基金资助:
    国家自然科学基金;广东省省级科技计划项目

OSKM induced c-Jun overexpression of liver cancer cell reprogramming

SU Ming1 HAN Shuo1 ZHANG Ming-zhi1 LI Rui-zhi1 WEI Shi-ruo1 ZENG Wo-tan2 LIU Jin-wen2 SHEN Li 1*   

  1. 1. Department of Cell Biology, Peking University Health Science Center, Beijing 100191, China, 2. Beijing Oriental Yamei Gene Science and Technology Research Institute Co. LTD, Beijing, 100078, China
  • Received:2016-11-07 Revised:2017-01-03 Online:2017-06-06 Published:2017-09-19
  • Contact: SHEN Li E-mail:shenli@bjmu.edu.cn

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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-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。

关键词: c-Jun, 诱导, 肿瘤干细胞, 重编程, 免疫印迹法, C3A细胞

Abstract:

Objective To establish an OSKM induced C3A-c-Jun of liver cancer reprogrammed cell line, and to explore the effects of exogenous c-Jun on liver cancer cell reprogramming. Methods OSKM was utilized to induce C3A-c-Jun to reprogram. Alkaline phosphatase staining, Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR), Western blotting, and immunofluorescence technology were used to identify cells, and establish the C3A-c-Jun-iCSCs. Results C3A-c-Jun-iCSCs cells formed dome-shaped clones, in which cells were small, closely packed and multi-layered. Alkaline phosphatase staining was positive. They expressed pluripotent markers Oct4, Sox2, Nanog at the mRNA level and express OCT4, and SOX2 at the protein level. C3A-c-Jun-iCSCs had obviously higher gene expression of Sox2 compared with C3A-iCSCs, whatever in exogenous expression or endogenous expression. C-Jun continually expressed in the process of reprogramming. Conclusion OSKM induces C3A/C3A-c-Jun of liver cancer reprogramming and built the C3A-i-CSCs/C3A-c-J-un-iCSCs. Exogenous c-Jun has a promoting effect on liver cancer cell reprogramming process by upregulating Sox2, which then can activate downloaded genes.

Key words: c-Jun, Induce, Cancer stem cell, Reprogramming, Western blotting, C3A cell