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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2018, Vol. 38 Issue (12): 1-6    DOI: 10.13523/j.cb.20181201
研究报告     
敲低ACTL6A通过Notch1信号通路促进早幼粒细胞分化 *
钟鹏强1,2,刘北忠1,2(),姚娟娟1,2,刘冬冬2,袁桢2,刘俊梅1,陈敏1,2,钟梁2,**()
1 重庆医科大学附属永川医院中心实验室 重庆 402160
2 重庆医科大学临床检验诊断学教育部重点实验室 重庆重点实验室 重庆 400016
Knock-down of ACTL6A Promote Differentiation of NB4 Cells via the Notch1 Signaling Pathway
ZHONG Peng-qiang1,2,LIU Bei-zhong1,2(),YAO Juan-juan1,2,LIU Dong-dong2,YUAN Zhen2,LIU Jun-mei1,CHEN Min1,2,ZHONG Liang2,**()
1 Central Laboratory,Yongchuan Hospital of Chongqing Medical University,Chongqing 402160, China
2 Key Laboratory of Chongqing,Key Laboratory of Medical Diagnostics of Ministry of Education,Department of Laboratory Medicine,Chongqing Medical University,Chongqing 400016,China
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摘要：

探讨ACTL6A在人类白血病NB4细胞分化中的作用及其相关机制。我们用ATRA人为诱导NB4细胞分化,Western blotting检测ACTL6A和CD11b的表达水平变化;敲低ACTL6A,通过瑞氏染色观察NB4细胞的形态学改变,Western blotting检测ACTL6A和CD11b的表达水平变化及其相关蛋白的表达水平;敲低同时用ATRA处理NB4细胞,用流式细胞术检测分化标志物CD11b的阳性率;免疫荧光检测ACTL6A在NB4细胞中的空间定位;结果显示NB4经敲低ACTL6A后,CD11b的蛋白水平表达升高;瑞氏染色观察到分化改变;免疫荧光检测到ACTL6A主要分布于细胞核;Western blotting 检测到Notch1,Hes1,Sox2蛋白表达水平明显下调。研究表明,敲低ACTL6A可以促进人类白血病NB4细胞分化;其机制涉及Notch1信号通路的抑制。
关键词： ACTL6A;  NB4细胞;  分化;  Notch1信号通路    
Abstract:

To explore the role of ACTL6A in NB4 cell differentiation and its related mechanisms.When we used ATRA to induce differentiation of NB4,the expression levels of ACTL6A and CD11b were detected by Western blotting.Morphological changes of NB4 cells were observed by Wright staining.The positive rate of CD11b was detected by flow cytometry.Immunofluorescence was used to detect location of ACTL6A in NB4 cells.The results showed that the protein expression of CD11b increased after ACTL6A was knocked down.Differentiation changes were observed by Wright staining.Immunofluorescence detection showed that ACTL6A was mainly distributed in the nucleus.The expression levels of Notch1,Hes1 and Sox2 proteins were significantly down-regulated,analysed by Western blotting. Therefore,the knockdown of ACTL6A can promote the differentiation of NB4 cells.The mechanism is involved in the inhibition of Notch1 signaling pathway.
Key words: ACTL6A    NB4 cells    Differentiation    Notch1 signal pathway
收稿日期: 2018-07-30 出版日期: 2019-01-10
ZTFLH:  Q813  
基金资助: * 国家自然科学基金(81772280)
通讯作者: 钟梁     E-mail: Liubeizhong@cqmu.edu.cn
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钟鹏强
刘北忠
姚娟娟
刘冬冬
袁桢
刘俊梅
陈敏
钟梁

引用本文:

钟鹏强,刘北忠,姚娟娟,刘冬冬,袁桢,刘俊梅,陈敏,钟梁. 敲低ACTL6A通过Notch1信号通路促进早幼粒细胞分化 *[J]. 中国生物工程杂志, 2018, 38(12): 1-6.

ZHONG Peng-qiang,LIU Bei-zhong,YAO Juan-juan,LIU Dong-dong,YUAN Zhen,LIU Jun-mei,CHEN Min,ZHONG Liang. Knock-down of ACTL6A Promote Differentiation of NB4 Cells via the Notch1 Signaling Pathway. China Biotechnology, 2018, 38(12): 1-6.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20181201    或    https://manu60.magtech.com.cn/biotech/CN/Y2018/V38/I12/1

图1  ACTL6A在ATRA诱导NB4细胞分化过程中表达下调
图2  敲低ACTL6A促进NB4细胞分化
图3  敲低ACTL6A增强ATRA诱导的NB4细胞的分化
图4  ACTL6A在细胞内的定位
图5  miR-302a可以下调ACTL6A 的表达
图6  敲低ACTL6A抑制Notch1信号通路,促进NB4细胞分化
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