三结构域蛋白21通过下调自噬相关蛋白4B抑制肝细胞癌细胞增殖的实验研究

李涛; 陈岺曦; 孙梁博; 闫小晶; 连继勤; 何凤田

doi:10.16016/j.1000-5404.201811167

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三结构域蛋白21通过下调自噬相关蛋白4B抑制肝细胞癌细胞增殖的实验研究

基础医学 | 更新时间：2026-01-09

- 三结构域蛋白21通过下调自噬相关蛋白4B抑制肝细胞癌细胞增殖的实验研究
- Tripartite motif 21 inhibits proliferation of hepatocellular carcinoma cells through down-regulating autophagy related 4B
- 第三军医大学学报 2019年41卷第9期页码：815-822
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)基础医学院生物化学与分子生物学教研室
  2.400037 重庆，陆军军医大学(第三军医大学)第二附属医院全军肿瘤研究所
- 作者简介：
  
  [何凤田，E-mail:hefengtian06@aliyun.comHE Fengtian, E-mail:hefengtian06@aliyun.com
- 基金信息：
  
  国家自然科学基金面上项目(81572375);重庆市自然科学基金面上项目(CSTC2018jcyjA2018)
- DOI：10.16016/j.1000-5404.201811167
  中图分类号： R394.3;R730.23;R735.7
- 收稿：2018-11-23，
  
  修回：2019-01-05，
  
  纸质出版：2019-05-15
- 稿件说明：
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李涛, 陈岺曦, 孙梁博, 等. 三结构域蛋白21通过下调自噬相关蛋白4B抑制肝细胞癌细胞增殖的实验研究[J]. 第三军医大学学报, 2019,41(9):815-822.

Tao LI, Linxi CHEN, Liangbo SUN, et al. Tripartite motif 21 inhibits proliferation of hepatocellular carcinoma cells through down-regulating autophagy related 4B[J]. Journal of Third Military Medical University, 2019, 41(9): 815-822.
李涛, 陈岺曦, 孙梁博, 等. 三结构域蛋白21通过下调自噬相关蛋白4B抑制肝细胞癌细胞增殖的实验研究[J]. 第三军医大学学报, 2019,41(9):815-822. DOI： 10.16016/j.1000-5404.201811167.

Tao LI, Linxi CHEN, Liangbo SUN, et al. Tripartite motif 21 inhibits proliferation of hepatocellular carcinoma cells through down-regulating autophagy related 4B[J]. Journal of Third Military Medical University, 2019, 41(9): 815-822. DOI： 10.16016/j.1000-5404.201811167.

摘要

目的

探讨三结构域蛋白21(tripartite motif 21，TRIM21)抑制肝细胞癌(hepatocellular carcinoma，HCC)细胞增殖的可能机制。

方法

采用TCGA数据库的样本数据分析TRIM21在HCC组织中的表达水平；Western blot检测HCC细胞系中TRIM21表达情况，在HepG2细胞中过表达TRIM21，并在SMMC7721细胞中敲低TRIM21；CCK-8和细胞克隆形成实验分别检测过表达和敲低TRIM21对HepG2和SMMC7721细胞增殖的影响；流式细胞术检测细胞凋亡的情况；Western blot检测TRIM21对自噬相关蛋白4B(autophagy related 4B，ATG4B)水平的影响；分别在HepG2细胞中下调ATG4B表达和在SMMC7721细胞中过表达ATG4B，CCK-8检测细胞增殖变化，流式细胞术检测细胞周期变化。

结果

TRIM21在HCC组织中的表达水平较正常肝组织显著下调(

=49，

0.05)；在HepG2细胞中过表达TRIM21能显著降低细胞存活率(

0.01)、抑制细胞增殖(

0.01)，在SMMC7721细胞中下调TRIM21的表达能显著增加细胞存活率(

0.05)、促进细胞增殖(

0.05)。过表达或敲低TRIM21对HCC细胞凋亡影响不明显；TRIM21在HCC细胞中可以抑制ATG4B的表达；而在HCC细胞中过表达或下调ATG4B也可以发挥促进或抑制细胞增殖的效果(

0.05)，ATG4B影响HCC细胞增殖的机制主要是改变细胞周期进程。

结论

TRIM21在HCC中可通过下调ATG4B而抑制细胞增殖。

Abstract

Objective

To investigate the possible mechanism of tripartite motif 21 (TRIM21)-triggered inhibition of cell proliferation in hepatocellular carcinoma (HCCs) cells.

Methods

Gene expression analysis by TGGA database was employed to get the TRIM21 expression level of HCC tissues and normal tissues. Western blotting was used to determine the protein level of TRIM21 in HCC cell lines HepG2 and SMMC7721. The TRIM21 level was up-regulated in HepG2 cells by TRIM21 expression plasmid

and down-regulated in SMMC7721 cells by TRIM21 siRNA

respectively. The effects of over-expression and knockdown of TRIM21 on cell proliferation were observed with CCK-8 assay and plate colony formation assay. Flow cytometry was employed to detect apoptotic rate. The expression level of autophagy related 4B (ATG4B) was measured by Western blotting. After ATG4B was down-regulated in HepG2 cells and up-regulated in SMMC7721 cells

cell viability and cell cycle were observed with CCK-8 assay and flow cytometry.

Results

TRIM21 was dramatically down-regulated in HCC tissues compared to normal liver tissues (

=49

0.05). Over-expression of TRIM21 significant

ly inhibited the viability and proliferation of HepG2 cells (both

0.01)

and knockdown of TRIM21 markedly enhanced the viability and proliferation in SMMC7721 cells (both

0.05). However

over-expression and knockdown of TRIM21 over- and down-expression had no obvious effects on the cell apoptosis. In HCC cells

up-regulation of TRIM21 suppressed ATG4B expression

While

up- or down-regulation of ATG4B could enhance or repress the cell proliferation (

0.05). ATG4B affected cell proliferation mainly through regulating cell cycle progression.

Conclusion

TRIM21 inhibits cell proliferation by down-regulating ATG4B in HCC cells.

关键词

Keywords

references

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