Effects of silencing APN gene on proliferation, invasion and apoptosis of human glioma U251 cells

Yunpeng DONG; Yuan CHENG; Jin CHEN

doi:10.16016/j.1000-5404.201912238

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Effects of silencing APN gene on proliferation, invasion and apoptosis of human glioma U251 cells

Basic Medicine | 更新时间：2026-01-09

- Effects of silencing APN gene on proliferation, invasion and apoptosis of human glioma U251 cells
- Journal of Third Military Medical University Vol. 42, Issue 9, Pages: 908-914(2020)
- 作者机构：
  
  400010 重庆, 重庆医科大学附属第二医院神经外科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81401505)
- DOI：10.16016/j.1000-5404.201912238
  CLC： R394.3;R730.23;R730.264
- Received：25 December 2019，
  
  Revised：2020-02-08，
  
  Published：15 May 2020
- 稿件说明：
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Yunpeng DONG, Yuan CHENG, Jin CHEN. Effects of silencing APN gene on proliferation, invasion and apoptosis of human glioma U251 cells[J]. Journal of Third Military Medical University, 2020, 42(9): 908-914.
DOI：

Yunpeng DONG, Yuan CHENG, Jin CHEN. Effects of silencing APN gene on proliferation, invasion and apoptosis of human glioma U251 cells[J]. Journal of Third Military Medical University, 2020, 42(9): 908-914. DOI： 10.16016/j.1000-5404.201912238.

摘要

目的

探究aminopeptidase N(APN)对胶质瘤细胞U251增殖能力、迁移侵袭能力和凋亡等生物学行为的影响及可能的作用机制。

方法

通过RNA干扰技术沉默胶质瘤细胞U251中APN的表达后

运用RT-qPCR的方法检测APN mRNA表达，采用Western blot检测APN蛋白表达水平，CCK-8检测各组U251细胞的增殖活性；采用流式细胞仪检测细胞凋亡和细胞周期；采用细胞划痕实验检测U251细胞的迁移能力，Transwell实验检测细胞侵袭能力；采用Western blot检测STAT3通路蛋白t-STAT3、p-STAT3、Bcl-2和MMP-2的蛋白表达水平。

结果

与空白对照组和阴性对照组相比，沉默U251中APN的表达后APN在基因和蛋白表达水平下降(

0.01)，沉默APN后U251的增殖活性明显下降(

0.05)，细胞周期停滞在G

期，并且凋亡率明显增加(

0.01)；另外沉默APN的表达后U251细胞的迁移能力和侵袭能力明显下降(

0.01)；沉默APN后STAT3磷酸化水平下降，引起下游蛋白Bcl-2和MMP-2等蛋白表达下降(

0.05)。

结论

沉默APN可通过STAT3信号途径抑制胶质瘤细胞U251增殖、迁移和侵袭，诱导凋亡。

Abstract

Objective

To determine the effect of aminopeptidase N (APN) on the proliferation

invasion

and apoptosis in human glioma U251 cells and to investigate the possible mechanism.

Methods

After APN gene expression was silenced in glioma cells U251 by RNAi

its expression at mRNA and protein levels was detected by RT-qPCR and Western blotting

respectively. Then

CCK-8 assay

flow cytometry

wound healing test and transwell assay were used to detect the proliferation ability

cell cycle and apoptosis

migration and invasion activity in the U251 cells. The protein expression of STAT3 signal pathway proteins t-STAT3

p-STAT3

MMP-2 and Bcl-2 was measured by Western blotting.

Results

Compared with blank control and NC-siRNA cells

the expression of APN was decreased at mRNA and protein levels in APN-siRNA cells (

0.01). Silencing of APN gene also

resulted in inhibited proliferation ability (

0.05)

cell cycle arrested at G

phase

enhanced apoptotic rate (

0.01)

and decreased migration and invasion abilities (

0.01) in U251 cells. What's more

the inhibited expression of p-STAT3 caused the decreased expression of Bcl-2 and MMP-2 (all

0.05).

Conclusion

Silencing of APN could inhibit the proliferation

migration and invasion

and induce apoptosis in glioma U251 cells

via

STAT3 signaling pathway.

关键词

Keywords

references

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