上海交通大学学报(医学版) ›› 2017, Vol. 37 ›› Issue (10): 1327-.doi: 10.3969/j.issn.1674-8115.2017.10.003

• 论著(基础研究) • 上一篇    下一篇

DOT1L-long enhances breast cancer metastasis#br#

DING Xiao-kai*, FU Yin-kun*, MAN MOHAN   

  1. Department of Biochemistry and Molecular Cell Biology, Basic Medicine Faulty of Shanghai Jiao Tong University, Shanghai 200025, China
  • 出版日期:2017-10-28 发布日期:2017-11-01
  • 通讯作者: ?MAN MOHAN,电子信箱:man@shsmu.edu.cn
  • 作者简介:丁小凯(1990—),硕士生;电子信箱:dingxk@outlook.com。付寅坤(1986—),助理实验师,硕士;电子信箱:yinkunfu@shsmu.edu.cn。* 共同第一作者
  • 基金资助:
    国家自然科学基金(31471206);上海市科学技术委员会基础科研项目 (14JC1404000)

DOT1L-long enhances breast cancer metastasis#br#

DING Xiao-kai*, FU Yin-kun*, MAN MOHAN   

  1. Department of Biochemistry and Molecular Cell Biology, Basic Medicine Faulty of Shanghai Jiao Tong University, Shanghai 200025, China
  • Online:2017-10-28 Published:2017-11-01
  • Supported by:
    National Natural Science Foundation of China, 31471206; Basic Science Foundation of Science and Technology Commission of Shanghai Municipality, 14JC1404000

摘要:  Objective · To investigate the histone methyltransferase capability of DOT1L-long form and its role in breast cancer metastasis.  Methods · The existence of DOT1L-long form was confirmed by PCR, and the mRNA level of DOT1L was tested by real-time PCR. In HEK293T cells in which DOT1L canonical and DOT1L-long were overexpressed respectively, Western blotting was used to test the expression level of DOT1L and the histone methyltransferase capability. In the MCF10A cell line with inducible expression of DOT1L-long, real-time PCR was used to detect the mRNA level of epithelial-mesenchymal transition (EMT) marker, and transwell assay was used to detect the migration of breast cancer cells in which the expression level of DOT1L is low or high.  Results · PCR demonstrated the existence of DOT1L-long form, and real-time PCR showed it widely exists in HCT116, T98G, MCF10A cells, etc. Western blotting showed the expression of DOT1L-long form and its H3K79 methyltransferase activity. In MCF10A cells in which overexpressed canonical DOT1L and DOT1L-long, mRNA levels of N-cadherin and fibronectine increased. Transwell showed canonical DOT1L and DOT1L-long both substantially increased the migration of breast cancer cells.  Conclusion · The existence of DOT1L-long was confirmed and investigated, which is 202 amino acids longer than the canonical DOT1L, and is coded by a new exon, located between exon 27 and 28. Further, the DOT1L-long has H3K79 methyltransferase activity, and is able to promote breast cancer metastasis.

关键词: DOT1L, histone methyltransferase, long form, breast cancer, metastasis

Abstract:

 Objective · To investigate the histone methyltransferase capability of DOT1L-long form and its role in breast cancer metastasis.  Methods · The existence of DOT1L-long form was confirmed by PCR, and the mRNA level of DOT1L was tested by real-time PCR. In HEK293T cells in which DOT1L canonical and DOT1L-long were overexpressed respectively, Western blotting was used to test the expression level of DOT1L and the histone methyltransferase capability. In the MCF10A cell line with inducible expression of DOT1L-long, real-time PCR was used to detect the mRNA level of epithelial-mesenchymal transition (EMT) marker, and transwell assay was used to detect the migration of breast cancer cells in which the expression level of DOT1L is low or high.  Results · PCR demonstrated the existence of DOT1L-long form, and real-time PCR showed it widely exists in HCT116, T98G, MCF10A cells, etc. Western blotting showed the expression of DOT1L-long form and its H3K79 methyltransferase activity. In MCF10A cells in which overexpressed canonical DOT1L and DOT1L-long, mRNA levels of N-cadherin and fibronectine increased. Transwell showed canonical DOT1L and DOT1L-long both substantially increased the migration of breast cancer cells.  Conclusion · The existence of DOT1L-long was confirmed and investigated, which is 202 amino acids longer than the canonical DOT1L, and is coded by a new exon, located between exon 27 and 28. Further, the DOT1L-long has H3K79 methyltransferase activity, and is able to promote breast cancer metastasis.

Key words: DOT1L, histone methyltransferase, long form, breast cancer, metastasis