Effects of circTRIM33-12 on proliferation,apoptosis and epithelial-mesenchymal transition of brain glioma cells by regulating miR-191/DAB2 axis

CHEN Bing; FENG Hao; ZOU Cheng-gong; TANG Hui

doi:10.11659/jjssx.11E022142

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Effects of circTRIM33-12 on proliferation,apoptosis and epithelial-mesenchymal transition of brain glioma cells by regulating miR-191/DAB2 axis

Basic Research | 更新时间：2024-01-22

- Effects of circTRIM33-12 on proliferation,apoptosis and epithelial-mesenchymal transition of brain glioma cells by regulating miR-191/DAB2 axis
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 33, Issue 1, Pages: 36-42(2024)
- 作者机构：
  
  南充市中心医院神经外科，四川南充 637000
- 作者简介：
- 基金信息：
- DOI：10.11659/jjssx.11E022142
  CLC： R739.41
- Published：25 January 2024，
  
  Received：25 November 2022，
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陈兵,冯浩,邹成功等.circTRIM33-12调控miR-191/DAB2轴对脑胶质瘤细胞增殖、凋亡及上皮-间质转化的影响[J].局解手术学杂志,2024,33(01):36-42.

CHEN Bing,FENG Hao,ZOU Cheng-gong,et al.Effects of circTRIM33-12 on proliferation,apoptosis and epithelial-mesenchymal transition of brain glioma cells by regulating miR-191/DAB2 axis[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2024,33(01):36-42.
陈兵,冯浩,邹成功等.circTRIM33-12调控miR-191/DAB2轴对脑胶质瘤细胞增殖、凋亡及上皮-间质转化的影响[J].局解手术学杂志,2024,33(01):36-42. DOI： 10.11659/jjssx.11E022142.

CHEN Bing,FENG Hao,ZOU Cheng-gong,et al.Effects of circTRIM33-12 on proliferation,apoptosis and epithelial-mesenchymal transition of brain glioma cells by regulating miR-191/DAB2 axis[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2024,33(01):36-42. DOI： 10.11659/jjssx.11E022142.

摘要

目的

探讨circTRIM33-12通过miR-191/DAB2轴对脑胶质瘤细胞增殖、凋亡及上皮-间质转化（EMT）的影响及其作用机制。

方法

RT-qPCR法检测circTRIM33-12、miR-191和DAB2在脑胶质瘤细胞CHG-5和人正常脑胶质上皮细胞HEB中的表达。将培养的CHG-5细胞进行分组：siRNA NC组、circTRIM33-12 siRNA组、DAB2 siRNA组；mimics NC组、miR-191 mimics组；circTRIM33-12 WT+mimics NC组、circTRIM33-12 WT+miR-191 mimics组、circTRIM33-12 MUT+mimics NC组、circTRIM33-12 MUT+miR-191 mimics组；inhibitor NC组、miR-191 inhibitor组；pcDNA+mimics NC组、pcDNA-TRIM33-12+mimics NC组、pcDNA+miR-191 mimics组、pcDNA-TRIM33-12+miR-191 mimics组；DAB2 WT+mimics NC组、DAB2 WT+miR-191 mimics组、DAB2 MUT+mimics NC组、DAB2 MUT+miR-191 mimics组。CCK-8实验检测circTRIM33-12、miR-191和DAB2的表达对CHG-5细胞增殖能力的影响；流式细胞仪检测circTRIM33-12、miR-191和DAB2的表达对CHG-5细胞凋亡的影响；Western blot检测circTRIM33-12、miR-191和DAB2的表达对CHG-5细胞EMT的影响。TargetScan数据库分析miR-191与DAB2、circTRIM33-12的关系，双荧光素酶报告基因实验验证其关系。RT-qPCR法检测circTRIM33-12通过miR-191对DAB2表达的影响。

结果

与HEB细胞相比，CHG-5细胞中circTRIM33-12的表达下调(

0.01)，miR-191表达上调(

0.01)，DAB2表达下调(

0.01)。与siRNA NC组相比，circTRIM33-12 siRNA组和DAB2 siRNA组CHG-5细胞增殖活力、N-cadherin表达明显升高(

0.01)，细胞凋亡率、E-cadherin表达降低(

0.01)。circTRIM33-12靶向miR-191，miR-191靶向DAB2。与inhibitor NC组相比，miR-191 inhibitor组CHG-5细胞增殖活力、N-cadherin表达明显降低(

0.01)，细胞凋亡率E-cadherin表达升高(

0.01)。circTRIM33-12过表达通过miR-191抑制CHG-5细胞增殖与EMT。

结论

circTRIM33-12可能通过miR-191/DAB2轴调控脑胶质瘤细胞的增殖、凋亡与EMT。

Abstract

Objective

To investigate the effects and mechanism of circTRIM33-12 on proliferation，apoptosis and epithelial-mesenchymal transition （EMT） of brain glioma cells by miR-191/DAB2 axis.

Methods

The expressions of circTRIM33-12，miR-191 and DAB2 in brain glioma cell CHG-5 and human normal brain glial epithelial cells HEB were detected by RT-PCR. The cultured CHG-5 cells were divided into the siRNA NC group，the circTRIM33-12 siRNA group，the DAB2 siRNA group； the mimics NC group，the miR-191 mimics group； the circTRIM33-12 WT+mimics NC group，the circTRIM33-12 WT+miR-191 mimics group，the circTRIM33-12 MUT+mimics NC group，the circTRIM33-12 MUT+miR-191 mimics group； the inhibitor NC group，the miR-191 inhibitor group； the pcDNA+mimics NC group，the pcDNA-TRIM33-12+mimics NC group，the pcDNA+miR-191 mimics group，the pcDNA-TRIM33-12+miR-191 mimics group； the DAB2 WT+mimics NC group，the DAB2 WT+miR-191mimis group，the DAB2 MUT+mimics NC group，the DAB2 MUT+miR-191 mimis group. CCK-8 assay was used to detect the effects of the expressions of circTRIM33-12，miR-191 and DAB2 on the proliferation ability of CHG-5 cells； flow cytometry was used to detect the effects of the expressions of circTRIM33-12，miR-191 and DAB2 on the apoptosis of CHG-5 cells； Western blot was used to detect the effects of the expressions of circTRIM33-12，miR-191 and DAB2 on EMT of CHG-5 cells.TargetScan database was used to analyze the correlations among miR-191，circTRIM33-12 and DAB2，and dual luciferase reporter gene assay was used to verify their relationships； RT-qPCR was used to detect the effect of circTRIM33-12 on DAB2 expression through miR-191.

Results

Compared with HEB cells，the expression of circTRIM33-12 in CHG-5 cells was down-regulated （

0.01），the expression of miR-191 was up-regulated （

0.01），and the expression of DAB2 was down-regulated （

0.01）. Compared with the siRNA NC group，the proliferation activity and N-cadherin expression of CHG-5 cells in the circTRIM33-12 siRNA group and the DAB2 siRNA group were significantly increased （

0.01），while the apoptosis rate and E-cadherin expression were decreased （

0.01）. circTRIM33-12 targeted miR-191，and miR-191 targeted DAB2. Compared with the inhibitor NC group，the proliferation activity and N-cadherin expression of CHG-5 cells in the miR-191 inhibitor group were significantly decreased （

0.01），while the apoptosis rate and E-cadherin expression were increased （

0.01）.circTRIM33-12 overexpression inhibited CHG-5 cell proliferation and EMT through miR-191.

Conclusion

circTRIM33-12 may regulate the proliferation，apoptosis and EMT of brain glioma cells through the miR-191/DAB2 axis.

关键词

circTRIM33-12miR-191DAB2CHG-5细胞增殖上皮-间质转化

Keywords

circTRIM33-12miR-191DAB2CHG-5 cellsproliferationepithelial-mesenchymal transition

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