Long non-coding RNA CCAT2 promotes proliferation of non-small cell lung cancer cells by inhibiting transforming growth factor-β pathway

Jingjing ZHANG; Ruifang HUA; Yinghua JI; Min ZHANG; Weiwei LI; Qinghui YANG; Jin WANG; Haichuan YU; Ping LU; Xiangpeng WANG

doi:10.16016/j.1000-5404.201708038

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Long non-coding RNA CCAT2 promotes proliferation of non-small cell lung cancer cells by inhibiting transforming growth factor-β pathway

Basic Medicine | 更新时间：2026-01-09

- Long non-coding RNA CCAT2 promotes proliferation of non-small cell lung cancer cells by inhibiting transforming growth factor-β pathway
- Journal of Third Military Medical University Vol. 40, Issue 2, Pages: 122-129(2018)
- 作者机构：
  
  1.453003 河南新乡，河南省新乡医学院医学检验学院和分子诊断与医学检验技术河南省协同创新中心
  2.453100 河南卫辉，河南省新乡医学院第一附属医院肿瘤科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(31301135);the Natural Science Foundation of Henan Province(162300410211);the Henan Medical Science and Technology Development Program(201203068)
- DOI：10.16016/j.1000-5404.201708038
  CLC： R394.3;R730.23;R734.2
- Received：07 August 2017，
  
  Revised：2017-11-06，
  
  Published：30 January 2018
- 稿件说明：
移动端阅览
Jingjing ZHANG, Ruifang HUA, Yinghua JI, et al. Long non-coding RNA CCAT2 promotes proliferation of non-small cell lung cancer cells by inhibiting transforming growth factor-β pathway[J]. Journal of Third Military Medical University, 2018, 40(2): 122-129.
DOI：

Jingjing ZHANG, Ruifang HUA, Yinghua JI, et al. Long non-coding RNA CCAT2 promotes proliferation of non-small cell lung cancer cells by inhibiting transforming growth factor-β pathway[J]. Journal of Third Military Medical University, 2018, 40(2): 122-129. DOI： 10.16016/j.1000-5404.201708038.

摘要

目的

通过体内体外实验探讨结肠癌相关转录本2(colon cancer-associated transcript 2，CCAT2)对非小细胞肺癌(non-small cell lung cancer

NSCLC)细胞增殖的影响及其调控机制。

方法

利用shRNA干扰NSCLC细胞(A549和H1299) CCAT2表达。实时定量PCR (quantitative real-time reverse transcription PCR，qRT-PCR)检测CCAT2表达。肿瘤成球实验和CCK-8实验检测细胞增殖。蛋白印记法检测细胞增殖标记蛋白-细胞增殖核抗原-67 (antigen identified by monoclonal antibody

Ki-67)和增殖细胞核抗原(proliferating Cell Nuclear Antigen，PCNA)，TGF-β信号通路相关蛋白转化生长因子(transforming growth factor beta

TGF-β)，细胞信号转导分子Smad2(suppressor of mothers against decapentaplegic 2)和α-平滑肌肌动蛋白(alpha smooth muscle actin

α-SMA)的表达。

结果

NSCLC细胞系(A549、H1299、H1650和H358)中CCAT2的表达显著高于正常肺脏细胞系BEAS-2B(

0.01)。与对照组相比，转染CCAT2 shRNA后A549和H1299细胞中CCAT2的表达显著降低(

0.01)。A549和H1299细胞中CCAT2 shRNA组细胞增殖倍数明显低于对照组(

0.05)。与对照组相比，A549和H1299细胞中CCAT2 shRNA组细胞增殖标记蛋白Ki-67和PCNA表达显著降低(

0.01)，TGF-β信号通路相关蛋白TGF-β、Smad2和α-SMA表达显著升高(

0.01)。与对照组相比，CCAT2 shRNA组肿瘤体积显著变小(

0.01)，肿瘤组织中TGF-β信号通路相关蛋白TGF-β、Smad2和α-SMA表达显著升高(

0.01)。

结论

CCAT2可通过抑制TGF-β信号通路促进NSCLC细胞增殖和肿瘤生长。

Abstract

Objective

To explore the role of long non-coding RNA (lncRNA) colon cancer-associated transcript 2 (CCAT2) in regulating the proliferation of non-small cell lung cancer (NSCLC) cells.

Methods

The expression of CCAT2 in NSCLC cell lines A549 and H1299 was silenced by transfecting the cells with a short hairpin RNA (shRNA) targeting CCAT2. CCAT2 expression in the transfected cells was detected with quantitative real-time PCR (qRT-PCR)

and the cell proliferation was analyzed with CCK-8 assay and tumor cell sphere formation assay. The expression of antigen identified by monoclonal antibody Ki-67

proliferating cell nuclear antigen (PCNA)

transforming growth factor beta (TGF-β)

Smad2 and alpha smooth muscle actin (α-SMA) in the cells were detected with Western blotting.

Results

NSCLC cell lines A549

H1299

H1650 and H358 expressed significantly higher levels of CCAT2 than the normal lung cell line BEAS-2B (

0.01). Compared with the control cells

A549 and H1299 cells transfected with CCAT2 shRNA showed obviously decreased expression of CCAT2 (

0.01) and significantly lowered proliferation rates (

0.01). Transfection with CCAT2 shRNA resulted in significantly lowered expression of Ki-67 and PCNA but obviously enhanced expression of TGF-β pathway related proteins TGF-β

Smad2 and α-SMA (

0.01). In mouse models bearing A549 cel

l xenografts

the tumor volume was significantly smaller in CCAT2 shRNA group than in the control group

and the expression of TGF-β

Smad2 and α-SMA in the tumor tissues were significantly higher in CCAT2 shRNA group (

0.01).

Conclusion

CCAT2 promotes the proliferation of NSCLC cells by inhibiting TGF-β pathway both in vitro and in vivo.

关键词

Keywords

references

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