COMMD7基因活化PI3K/AKT信号通路促进HepG2细胞增殖转移的分子机制研究

顾勋鑫; 郑  璐; 尤  楠; 谭  晔; 邓昌林; 李  靖

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COMMD7基因活化PI3K/AKT信号通路促进HepG2细胞增殖转移的分子机制研究

基础研究 | 更新时间：2023-05-22

- COMMD7基因活化PI3K/AKT信号通路促进HepG2细胞增殖转移的分子机制研究
- COMMD7 gene promotes HepG2 proliferation and migration by activiating the PI3K/Akt signal pathway
- 局解手术学杂志 2016年第5期
- 作者机构：
  
  第三军医大学新桥医院肝胆外科,重庆,400037
- 作者简介：
- 基金信息：
  
  国家自然科学基金（81372561）;重庆市自然科学基金（CSTC2012jjA10079）
- DOI：
  中图分类号：
- 纸质出版日期：2016，
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顾勋鑫, 郑璐, 尤楠, 等. COMMD7基因活化PI3K/AKT信号通路促进HepG2细胞增殖转移的分子机制研究[J]. 局解手术学杂志, 2016,(5).

GU Xun-xin, ZHENG Lu, YOU Nan, et al. COMMD7 gene promotes HepG2 proliferation and migration by activiating the PI3K/Akt signal pathway[J]. null, 2016,(5).
顾勋鑫, 郑璐, 尤楠, 等. COMMD7基因活化PI3K/AKT信号通路促进HepG2细胞增殖转移的分子机制研究[J]. 局解手术学杂志, 2016,(5). DOI：

GU Xun-xin, ZHENG Lu, YOU Nan, et al. COMMD7 gene promotes HepG2 proliferation and migration by activiating the PI3K/Akt signal pathway[J]. null, 2016,(5). DOI：

摘要

[摘要]目的探索肝癌相关基因COMMD7促进HepG2细胞增殖转移的分子机制。方法设计针对COMMD7基因的特异性siRNA（small interference RNA）转染人类肝癌细胞株HepG2，作为干扰组(Si-HepG2)；并设置PTEN抑制剂处理组（Si+BpV-HepG2）；空白对照组（HepG2）；空载体对照组（N-HepG2）。qRT-PCR检测转染后各组COMMD7、PTEN基因mRNA的表达变化；Western blot检测各组COMMD7、PTEN、p-AKT和AKT蛋白的表达变化；MSP法检测PTEN基因甲基化水平；CKK8法检测细胞增殖能力；Transwell实验检测细胞在转染前后转移及侵袭能力的改变。结果siRNA-COMMD7 转染HepG2细胞后，qRT-PCR检测结果显示，与空白对照组及空载体对照组相比较干扰组COMMD7基因的表达量降低89.9%倍，PTEN基因的表达量升高2.841倍，差异均有统计学意义（P<0.05）；Western blot检测结果显示：siRNA干扰组COMMD7表达明显降低，PTEN表达升高；下游p-AKT表达受到显著抑制；PTEN抑制剂处理组PTEN表达受到显著抑制，下游p-AKT的表达明显增强；MSP法检测结果显示，干扰组与空白对照组及空载体对照组相比较，PTEN发生明显的去甲基化，表明抑制COMMD7表达可诱导PTEN去甲基化。CKK8实验结果显示，干扰组细胞较空白对照组及空载体对照组和PTEN抑制剂处理组增殖速度明显降低，差异均有统计学意义(P<0.05)；Transwell实验实验结果显示，干扰组穿膜细胞数（17.4±2.7），较空载体对照组（36.2±3.2）、空白对照组（41.6±4.5）及PTEN抑制剂处理组（47.6±1.8）明显减少，差异均有统计学意义（P<0.05）。结论siRNA干扰下调COMMD7基因的表达，可诱导HepG2细胞内抑癌基因PTEN的去甲基化，增加PTEN蛋白的表达而抑制PI3K/AKT信号通路，以降低HepG2细胞的增殖、迁移及侵袭能力。

Abstract

Abstract: ObjectiveTo investigate the mechanism of COMMD7

a hepatocellular carcinoma gene

promoting human hepatocellular carcinoma cell line (HePG2 cells) proliferation and migration.MethodsThe specific siRNA (small interference RNA) was designed for COMMD7 gene

siRNA transfected HepG2 cells was set as Si-HePG2 group

and the PTEN inhibitor treating group (Si+BpV-HePG2)

the control group (HePG2)

the empty vector group (HePG2 was infected empty vector

N-HePG2 group) were also set up.qRT-PCR was performed to evaluate the mRNA expression changes of COMMD7 and PTEN

and Western blot was performed to test the expression changes of COMMD7

PTEN

p-AKT

and AKT.MSP method was performed to detect methylation level;CCK8 was performed to test cell proliferation ability;Transwell was performed to detect the invasion of cells.ResultsThe results of qRT-PCR showed that the expression of COMMD7 gene in Si-HePG2 group was lower (0.101 times) than the control group and the N-HePG2 group

and the expression of PTEN gene was higher (2.841 times) than the control group and the N-HePG2 group

and all the results above were of statistically singificant difference (P<0.05).The results of Western blot showed that in Si-HePG2 group

the expression of COMMD7 reduced obviously

the expression of PTEN increased and the expression of p-AKT was significantly inhibited.In Si+Bpv-HePG2 group

the expression of PTEN was significantly inhibited and the expression of p-AKT was increased.The result of MSP showed that compared with the control group and the N-HePG2 group

Si-HePG2 group was more obviously demethylated

which indecated that the expression of COMMD7 gene could induce PTEN demethylation.The results of CCK8 showed that the proliferation in Si-HePG2 group was decreased compared with the control group

N-HePG2 group and Si+BpV-HePG2 group

and the difference was singificant (P＜0.05).The results of Transwell showed that the numbers of cell permeating septum in Si-HePG2 group

N-HePG2 group

control group and PTEN group were (17.4±2.7)

(36.2±3.2)

(41.6±4.5)and (47.6±1.8) respectively

which were obviously decreased with a significant difference (P<0.05).ConclusionSiRNA interference decreased the expression of COMMD7.It can induced the demethylation of PTEN gene in HePG2 and increase the expression of PTEN gene to inhibit PI3K/AKT signaling pathway

thus decreasing the proliferation

migration and invasion ability of HePG2 cells.

关键词

肝细胞癌COMMD7PI3K/AKT信号通路PTEN去甲基化

Keywords

hepatocellular carcinomaCOMMD7PI3k/AKT signaling pathwayPTENdemethylation

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