RNA结合蛋白TTP对肺腺癌细胞增殖影响的实验研究

罗勤利; 邓小垭; 董飞; 徐莉; 江涛

doi:10.16016/j.1000-5404.201811085

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RNA结合蛋白TTP对肺腺癌细胞增殖影响的实验研究

基础医学 | 更新时间：2026-01-09

- RNA结合蛋白TTP对肺腺癌细胞增殖影响的实验研究
- RNA binding protein tristetraprolin inhibits proliferation of lung adenocarcinoma cells in vitro
- 第三军医大学学报 2019年41卷第9期页码：851-858
- 作者机构：
  
  400016 重庆，重庆医科大学附属第一医院呼吸与危重症医学科
- 作者简介：
  
  [江涛，E-mail:j.tcq@163.comJIANG Tao, E-mail:j.tcq@163.com
- 基金信息：
  
  国家自然科学基金面上项目(81602430);重庆市科委基础与前沿一般项目(CSTC2016jcyjA0329)
- DOI：10.16016/j.1000-5404.201811085
  中图分类号： R341;R730.23;R734.2
- 收稿：2018-11-12，
  
  修回：2018-12-19，
  
  纸质出版：2019-05-15
- 稿件说明：
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罗勤利, 邓小垭, 董飞, 等. RNA结合蛋白TTP对肺腺癌细胞增殖影响的实验研究[J]. 第三军医大学学报, 2019,41(9):851-858.

Qinli LUO, Xiaoya DENG, Fei DONG, et al. RNA binding protein tristetraprolin inhibits proliferation of lung adenocarcinoma cells in vitro[J]. Journal of Third Military Medical University, 2019, 41(9): 851-858.
罗勤利, 邓小垭, 董飞, 等. RNA结合蛋白TTP对肺腺癌细胞增殖影响的实验研究[J]. 第三军医大学学报, 2019,41(9):851-858. DOI： 10.16016/j.1000-5404.201811085.

Qinli LUO, Xiaoya DENG, Fei DONG, et al. RNA binding protein tristetraprolin inhibits proliferation of lung adenocarcinoma cells in vitro[J]. Journal of Third Military Medical University, 2019, 41(9): 851-858. DOI： 10.16016/j.1000-5404.201811085.

摘要

目的

探讨TTP(tritetraprolin)过表达对肺腺癌细胞增殖影响及其可能的作用机制。

方法

构建带有四环素可调控(tetracycline-on，Tet-on)的TTP基因表达系统，利用慢病毒感染的方法转染肺腺癌H1975及1299细胞，并在强力霉素(doxycycline，Dox)的诱导下稳定表达TTP。用实时荧光定量PCR和Western blot鉴定TTP受Dox调控表达的效果。分别采用CCK-8法、Transwell、平板克隆形成实验、流式细胞术检测TTP过表达后对细胞增殖、迁移、集落形成能力、凋亡和细胞周期的影响。Western blot法检测TTP过表达对凋亡基因在蛋白水平表达的影响。

结果

成功构建受Dox诱导调控的TTP过表达肺腺癌细胞模型。与TTP低表达组比较，Dox诱导TTP过表达组明显抑制H1975、H1299细胞增殖、迁移能力(H1975 :

0.01

H1299:

0.001)及集落形成能力(

值均

0.0001)，且使H1975细胞周期被阻滞在S期(

0.01)，但TTP过表达不影响肺腺癌细胞凋亡。

结论

TTP过表达显著抑制肺腺癌细胞增殖、迁移、集落形成能力，使细胞周期被阻滞在S期，但TTP过表达不通过细胞凋亡途径影响肺腺癌细胞增殖和生长。

Abstract

Objective

To investigate the effect of tristetraprolin (TTP) on the proliferation of lung cancer cells and to explore the possible mechanism.

Methods

The human lung adenocarcinoma cells with tetracycline-inducible (Tet-on) expression regulatory system were established in H1975 and 1299 cells with the aid of lentivirus infection. Western blotting and q-PCR were used to confirm the overexpression of TTP in the cells after the doxycycline (Dox) treatment. Effect of TTP on the proliferation

migration and colony-forming capacity were detected by CCK-8 assay

Transwell test and colony forming unit test respectively. The cell apoptosis and cell cycle were measured by flow cytometry. The expression of cellular apoptosis-related proteins were determined by Western blotting.

Results

Lung adenocarcinoma cells with Dox-induced TTP overexpression were successfully established. Compared with the cells with low TTP expression

those with Dox-induced TTP overexpression siginicantly inhibited proliferation

migration(H1975:

0.01

H1299:

0.001)and colony-formation (all

0.0001) in lung adenocarcinoma cells H1975 and H1299. What's more

H1975 cells were arrested at S stage (

0.01). However

TTP overexpression exerted no effect on cell apoptosis.

Conclusion

TTP overexpression obviously inhibits proliferation

migration and colony-forming capacity

and arrests cell cycle at S stage in lung adenocarcinoma cells. But the mechanism is not due to the regulation of cell apoptosis.

关键词

Keywords

references

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