Effects of FBXL20 on proliferation in human non-small cell lung cancer A549 cells

Bohan ZHANG; Qingchen WU

doi:10.16016/j.1000-5404.201711238

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Effects of FBXL20 on proliferation in human non-small cell lung cancer A549 cells

Basic Medicine | 更新时间：2026-01-09

- Effects of FBXL20 on proliferation in human non-small cell lung cancer A549 cells
- Journal of Third Military Medical University Vol. 40, Issue 9, Pages: 795-800(2018)
- 作者机构：
  
  400016 重庆, 重庆医科大学附属第一医院胸心外科
- 作者简介：
- 基金信息：
  
  the Key Scientific Research Project of Chongqing Municipal Health Bureau(2012-0-015)
- DOI：10.16016/j.1000-5404.201711238
  CLC： R394.3;R730.23;R734.2
- Received：15 November 2017，
  
  Revised：2018-01-28，
  
  Published：15 May 2018
- 稿件说明：
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Bohan ZHANG, Qingchen WU. Effects of FBXL20 on proliferation in human non-small cell lung cancer A549 cells[J]. Journal of Third Military Medical University, 2018, 40(9): 795-800.
DOI：

Bohan ZHANG, Qingchen WU. Effects of FBXL20 on proliferation in human non-small cell lung cancer A549 cells[J]. Journal of Third Military Medical University, 2018, 40(9): 795-800. DOI： 10.16016/j.1000-5404.201711238.

摘要

目的

观察FBXL20(F-box and leucine rich repeat protein 20)对人非小细胞肺癌A549细胞生长的影响并初步探究其机制。

方法

采用慢病毒感染A549细胞

构建对照组(FBXL20-vector组)及干扰组(FBXL20-RNAi组)

通过Western blot验证慢病毒敲减效率。利用CCK-8及集落形成实验观察FBXL20的下调对A549细胞增殖能力的影响

流式细胞术检测细胞周期的变化

Western blot分析周期蛋白的改变

基因集富集分析(gene set enrichment analysis

GSEA)探究FBXL20可能参与的生物学过程。

结果

干扰组FBXL20的蛋白表达(0.36±0.02)显著低于对照组(0.58±0.01)(

0.01)。与对照组相比

干扰组的增殖能力增强(

0.05)

形成的集落数目也显著增加(100±20

53±6)(

0.05)

且停留在G

/M期的细胞比例明显降低(5.65±1.35

9.94±1.44)(

0.05)。Western blot结果显示Cyclin D1、ERK1/2蛋白表达无明显变化(

0.05)

CDK2、Cyclin E、CDK1、Cyclin A、Cyclin B1蛋白表达显著增加

而p-ERK1/2则显著降低(

0.05)。GSEA结果显示FBXL20的低表达与G

/M检查点呈正相关(

0.01)。

结论

下调FBXL20可以加快A549细胞G

/M期的进程

提高其生长能力。其机制可能是通过抑制p-ERK的激活

使得其下游周期蛋白CDK1、Cyclin A、Cyclin B1表达增加而实现的。

Abstract

Objective

To detect the effect of F-box and leucine rich repeat protein 20 (FBXL20) on the proliferation of human non-small cell lung cancer A549 cells and mine the underlying mechanism.

Methods

Lentiviral vector FBXL20-RNAi was used to transfect A549 cells to knock down the expression of FBXL20

and blank lentiviral vector (FBXL20-vector) was used for control cells.Knockdown efficiency was verified by Western blotting.CCK-8 assay and colony formation assay were used to determine the effects of silenced FBXL20 on the proliferation.Flow cytometry was performed to explore cell cycle alteration.Expression changes of cell cycle proteins were measured by Western blotting.Gene set enrichment analysis (GSEA) was conducted to investigate the biological process involved with FBXL20.

Results

The protein level of FBXL20 was remarkably lower in the interference cells than the control cells (0.36±0.02

0.58±0.01

0.01).Compared with the control cells

the interference cel

ls had greater proliferation ability (

0.05) and increased colony count (100±20

53±6

0.05)

and less cells arrested in G

/M phase (5.65±1.35

9.94±1.44

0.05).Western blot results showed there were no significant differences in protein levels of Cyclin D1 and ERK1/2 (

0.05)

and the protein levels of CDK2

Cyclin E

CDK1

Cyclin A and Cyclin B1 were notably increased while p-ERK1/2 was distinctly decreased (

0.05).GSEA revealed that low level of FBXL20 was positively correlated with G

/M checkpoint (

0.01).

Conclusion

Down-regulation of FBXL20 can expedite the G

/M phase of A549 cells and enhance the growth ability

which may due to the suppression of p-ERK activation

and therefore enhancement of the expression of its downstream cell cycle proteins CDK1

Cyclin A and Cyclin B1.

关键词

Keywords

references

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