长非编码RNAZ38对BT549细胞多能因子表达调控的初步研究  

Primary Study on the Regulation of Pluripotent Factor Expression of BT549 Cells with Long Non-Coding RNA Z38

作　　者：孙园园[1] 刘斌[1] 欧阳婧[1] 莫林波 唐政山 乔小银 李放军[2] 杨寅柯[1] 

机构地区：[1]湖南大学生物学院,长沙410082 [2]湖南省人民医院社会医学部,长沙410002

出　　处：《中国细胞生物学学报》2017年第8期1041-1048,共8页Chinese Journal of Cell Biology

基　　金："985工程"专项资金(批准号:531109020011)资助的课题~~

摘　　要：该文旨在研究长非编码RNA Z38对BT549细胞中多能因子表达的影响。通过流式细胞术和实时荧光定量PCR(Real-time quantitative polymerase chain reaction,RT-q PCR)进行Z38与CD44+CD24–/low亚群细胞相关性分析,并利用慢病毒Z38-sh RNA感染BT549细胞以干扰Z38表达,通过RT-qPCR和蛋白质印迹实验检测多能因子表达水平。结果显示,Z38表达水平与CD44+CD24–/low亚群细胞比例存在差异性,以BT549细胞Z38表达水平和CD44+CD24–/low比例相对较高,干扰BT549细胞Z38表达水平后,CD44+CD24–/low比例下降,NANOG和SOX2 m RNA和蛋白质水平皆被下调,而OCT4 m RNA和蛋白质水平无明显变化。干扰Z38影响BT549多能因子的表达,尤其可下调与乳腺癌干细胞分化密切相关的NANOG和SOX2基因表达,提示它可能与乳腺癌干细胞的分化有关。Z38 is a newly discovered long non-coding RNA that promotes the formation of breast cancer, but the specific mechanism is unclear. The aim of this study was to investigate the effect of Z38 on the expression of pluripotency gene NANOG, SOX2 and OCT4 in BT549 cells. The correlation between Z38 and CD44^＋CD24^-/lowsubpopulations was analyzed by fl ow cytometry and RT-q PCR（Real-time quantitative polymerase chain reaction）. Z38-sh RNA was used to interfere Z38 level in BT549 cells. The m RNA and protein levels of NANOG, SOX2 and OCT4 were detected by RT-q PCR and Western blot. The results showed that the expression levels of Z38 and CD44^＋CD24^-/low subpopulations were different in MCF-7, MDA-MB-453 and BT549 breast cancer cells. The ratio of CD44^＋CD24^-/low subpopulations in BT549 cells was relatively high, and the percentage of CD44^＋CD24^-/low subgroups decreased in BT549 cells after interference of Z38. The results also showed that the genes NANOG and SOX2 were down-regulated at m RNA and protein levels, while the gene OCT4 had no significant change in m RNA and protein level. Interference of Z38 affected the expression of BT549 pluripotency factor, especially downregulating the NANOG and SOX2 genes closely related to the differentiation of breast cancer stem cells, suggesting that it may be involved in regulating the differentiation of breast cancer stem cells.

关 键 词：Z38 乳腺癌 NANOG SOX2 OCT4 CD44^+CD24^–/low 

分 类 号：R737.9[医药卫生—肿瘤]