OSKM诱导过表达c-Jun肝癌细胞重编程  

OSKM induced c-Jun overexpression of liver cancer cell reprogramming

作　　者：苏铭[1] 韩烁[1] 张明智[1] 李睿智[1] 魏示若 曾沃坦 刘进稳 沈丽[1] SU Ming HAN Shuo ZHANG Ming-zhi LI Rui-zhi WEI Shi-ruo ZENG Wo-tan LIU Jin-wen SHEN Li(Department of Cell Biology, Peking University Health Science Center, Beijing 100191, China Beijing Oriental Yamei Gene Science and Technology Research Institute Co. LTD, Beijing, 100078, China)

机构地区：[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

出　　处：《解剖学报》2017年第3期296-302,共7页Acta Anatomica Sinica

基　　金：国家自然科学基金(81572313);广东省省级科技计划项目(2015B020229002)

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。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 ceils 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.

关 键 词：C-JUN 诱导 肿瘤干细胞 重编程 免疫印迹法 C3A细胞 

分 类 号：R735.7[医药卫生—肿瘤]