RS4651通过上调SMAD7抑制小鼠肝细胞AML12的EMT作用

doi:10.13523/j.cb.2103067

中国生物工程杂志

2021, Vol. 41

Issue (7): 1-9 DOI: 10.13523/j.cb.2103067

研究报告

RS4651通过上调SMAD7抑制小鼠肝细胞AML12的EMT作用

李世荣¹,陈阳琴¹,张春盼^1,²,齐文杰^1,^*(

)

1 首都医科大学附属北京友谊医院北京 100050
2 国家消化系统疾病临床医学研究中心北京 100050

RS4651 Inhibits the EMT of Mouse Hepatocyte AML12 via Upregulating SMAD7

LI Shi-rong¹,CHEN Yang-qin¹,ZHANG Chun-pan^1,²,QI Wen-jie^1,^*(

)

1 Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
2 National Clinical Research Center for Digestive Disease, Beijing 100050, China

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摘要：

目的: 研究RS4651对小鼠肝细胞AML12的EMT、细胞增殖以及迁移能力的影响和作用机制。方法: 以不同浓度的RS4651干预小鼠AML12细胞,通过Western blot和RT-PCR法检测RS4651各浓度组及空白对照组细胞EMT指标E-cadherin、N-cadherin、Vimentin的表达水平。RNA-sequencing确定RS4651的作用通路及互作网络的关键性节点基因。应用SMAD7-siRNA构建SMAD7敲低的AML12细胞模型,细胞分为对照组、RS(60 μmol/L)组、SMAD7-siRNA组以及SMAD7-siRNA+RS(60 μmol/L)组,应用Western blot和RT-PCR法分别检测各组细胞EMT指标。进一步应用细胞增殖MTS法和Transwell小室迁移法检测各组细胞的增殖和迁移能力,比较各组间差异。结果: RS4651呈浓度依赖性抑制AML12细胞EMT,TGF-β1信号通路中的SMAD7是RS4651发挥作用的关键性节点。敲低SMAD7后,RS4651对AML12细胞EMT、增殖和迁移的抑制作用减弱。结论: RS4651可以通过上调SMAD7抑制小鼠肝细胞EMT、增殖和迁移。

关键词： RS4651; SMAD7; EMT; 增殖; 迁移

Abstract:

Objective: To investigate the effect of RS4651 on the EMT of hepatic cell AML12 in mice and the potential mechanism.Methods: AML12 cells were treated with RS4651 at different concentrations. Western blot and RT-PCR was used to detect the expression of E-cadherin, N-cadherin and Vimentin of RS4651 treatment groups and control group to investigate the effect of RS4651 on EMT of AML12 cells and SMAD7-Knockdown AML12 cells. RNA-sequencing identified the key node genes in the signaling pathway and the interaction network of RS4651. The proliferation and migratory effects of RS4651 treatment on AML12 cells with or without silencing by SMAD7-siRNA.Results: RS4651 could significantly upregulate the expression of E-cadherin and downregulated the expression of N-cadherin and Vimentin in a concentration-dependent manner. RNA-sequencing data showed that the target gene was SMAD7 in TGF-β1 signalling pathway. The expression of E-cadherin was relatively decreased in the SMAD7-siRNA+RS (60 μmol/L) group compared with the RS (60 μmol/L) group, while the expression of N-cadherin and Vimentin were relatively increased, and the proliferation and migration of AML12 cells were also increased.Conclusion: RS4651 can inhibit EMT, proliferation and migration of mice hepatocyte AML12 cells through SMAD7.

Key words: RS4651 SMAD7 EMT Proliferation Migration

收稿日期: 2021-03-26 出版日期: 2021-08-03

ZTFLH:

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通讯作者: 齐文杰 E-mail: qwj02@126.com

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引用本文:

李世荣,陈阳琴,张春盼,齐文杰. RS4651通过上调SMAD7抑制小鼠肝细胞AML12的EMT作用[J]. 中国生物工程杂志, 2021, 41(7): 1-9.

LI Shi-rong,CHEN Yang-qin,ZHANG Chun-pan,QI Wen-jie. RS4651 Inhibits the EMT of Mouse Hepatocyte AML12 via Upregulating SMAD7. China Biotechnology, 2021, 41(7): 1-9.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2103067 或 https://manu60.magtech.com.cn/biotech/CN/Y2021/V41/I7/1

表1 引物序列

图1 RS4651呈浓度依赖性抑制小鼠AML12细胞的EMT

图2 转录组测序证实RS4651抑制AML12细胞EMT的作用通路和靶点

图3 RS4651对SMAD7抑制AML12细胞EMT作用

图4 RS4651抑制AML12细胞的增殖能力

图5 RS4651通过SMAD7抑制AML12细胞的迁移能力

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