miR-185通过调控E2F6抑制肺鳞癌细胞增殖和侵袭

曹燕飞; 任睿; 杨晔; 罗向晖; 王水利

doi:10.16016/j.1000-5404.201607118

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miR-185通过调控E2F6抑制肺鳞癌细胞增殖和侵袭

基础医学 | 更新时间：2020-08-05

- miR-185通过调控E2F6抑制肺鳞癌细胞增殖和侵袭
- MicroRNA-185 inhibits proliferation and invasion of lung squamous cells via regulating E2F6
- 第三军医大学学报 2017年39卷第2期页码：123-129
- 作者机构：
  
  1.710068 西安, 陕西省人民医院: 胸外科
  2.710048 西安, 陕西省新安中心医院妇产科
  3.710068 西安, 陕西省人民医院: 呼吸内科
- 作者简介：
  
  任睿, E-mail: cyf.521@qq.com Ren Rui, E-mail: cyf.521@qq.com
- 基金信息：
  
  陕西省自然科学基金面上项目(2014JM2-8166)
- DOI：10.16016/j.1000-5404.201607118
  中图分类号： R394.3;R730.23;R734.2
- 纸质出版日期：2017-1-30，
  
  收稿日期：2016-7-18，
  
  录用日期：2016-9-7
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曹燕飞, 任睿, 杨晔, 等. miR-185通过调控E2F6抑制肺鳞癌细胞增殖和侵袭[J]. 第三军医大学学报, 2017,39(2):123-129.

Yanfei Cao, Rui Ren, Ye Yang, et al. MicroRNA-185 inhibits proliferation and invasion of lung squamous cells via regulating E2F6[J]. J Third Mil Med Univ, 2017,39(2):123-129.
曹燕飞, 任睿, 杨晔, 等. miR-185通过调控E2F6抑制肺鳞癌细胞增殖和侵袭[J]. 第三军医大学学报, 2017,39(2):123-129. DOI： 10.16016/j.1000-5404.201607118.

Yanfei Cao, Rui Ren, Ye Yang, et al. MicroRNA-185 inhibits proliferation and invasion of lung squamous cells via regulating E2F6[J]. J Third Mil Med Univ, 2017,39(2):123-129. DOI： 10.16016/j.1000-5404.201607118.

摘要

目的

探讨微小RNA-185(microRNA-185， miR-185)对肺鳞癌细胞增殖和侵袭能力的作用及其可能的作用机制。

方法

人肺鳞癌细胞系(NCI-H520、NCI-H226)以及正常肺上皮细胞(BEAS-2B)为实验用细胞； qRT-PCR检测miR-185表达量；miR-NC(对照)和miR-185 mimics(miR-185类似物)利用Turbofect进行转染；Western blot检测E2F6的蛋白表达量；双荧光素酶报告基因检测miR-185与E2F6的靶向调控关系；MTT检测细胞的增殖能力；细胞侵袭试剂盒检测细胞的侵袭能力；构建pcDNA.3.1-E2F6重组载体，并转染肺鳞癌细胞过表达E2F6。

结果

miR-185表达量在肺鳞癌细胞中显著降低(

＜0.01)。miR-185靶向抑制E2F6基因(

＜0.05)。转染miR-185 mimics显著上调miR-185的细胞含量(

＜0.01)并抑制E2F6蛋白表达(

＜0.01)，肺鳞癌细胞增殖能力(

＜0.05或

＜0.01)和侵袭能力(

＜0.05)亦显著受抑。转染pcDNA.3.1-E2F6到miR-185含量升高的细胞，E2F6蛋白表达量显著升高(

＜0.01)且细胞增殖 (

＜0.01)和侵袭能力(

＜0.05)上升。

结论

miR-185可以通过靶向E2F6抑制肺鳞癌细胞增殖和侵袭。

Abstract

Objective

To determine the effect of microRNA-185 (miR-185) on the proliferation and invasion of lung squamous cells and investigate the possible underlying mechanism.

Methods

Human lung squamous cancer cell lines (NCI-H520 and NCI-H226) and normal lung cell line (BEAS-2B) were used in the research. The miR-185 expression was detected by quantitative real-time PCR (qRT-PCR) in the above cells. The targeted relationship between miR-185 and E2F6 was detected by dual luciferase reporter assay. After MiRNA-NC and miR-185 mimics were transfected into the cells by using Turbofect

the protein expression of E2F6 was measured by Western blot assay

cell proliferation was detected by MTT assay

and cells invasion was measured by cell invasion kit. pcDNA.3.1-E2F6 recombinant plasmid was constructed

and transfected into lung squamous cells to over-express E2F6.

Results

miR-185 expression was significantly decreased in the lung squamous cells (

＜0.01). miR-185 directly targeted E2F6 gene (

＜0.05). miR-185 mimics transfection markedly up-regulated the content of miR-185 (

＜0.01) and inhibited the protein expression of E2F6 (

＜0.01)

and obviously suppressed the cell proliferation (

＜0.05 or

＜0.01) and invasion (

＜0.05) of lung squamous cells. Transfection of pcDNA.3.1-E2F6 resulted in the over-expression of miR-185

the up-regulation of E2F6 protein (

＜0.01)

and thus significantly promoted the cell proliferation (

＜0.01) and invasion (

＜0.05).

Conclusion

miR-185 inhibits the proliferation and invasion of the lung squamous cells

via

targeting E2F6.

关键词

肺鳞癌miR-185E2F6增殖侵袭

Keywords

squamous cell lung carcinomamiR-185E2F6proliferationinvasion

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