RS4651 Inhibits the EMT of Mouse Hepatocyte AML12 via Upregulating SMAD7

doi:10.13523/j.cb.2103067

China Biotechnology

2021, Vol. 41

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

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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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

Received: 26 March 2021 Published: 03 August 2021

ZTFLH:

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Corresponding Authors: Wen-jie QI E-mail: qwj02@126.com

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Cite this article:

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.

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

Table 1 Q-RT-PCR primers for E-cadherin, N-cadherin, vimentin, GAPDH

Fig.1 RS4651 inhibited EMT of AML12 cells in a concentration-dependent manner (a)The effect of RS4651 on the protein expression of EMT index in AML12 cells (b) The effect of RS4651 on the mRNA expression of EMT index in AML12 cells. ^* P<0.05,^** P<0.01

Fig.2 Transcriptome sequencing confirmed the pathway and target of RS4651’s inhibitory effect on EMT in AML12 cells (a) (b) The number of differentially expressed genes (c) Kegg enrichment analysis of the top 20 most significant cell component signals in differentially expressed genes (d) STRING analyzes the interaction network of nine differentially expressed genes in the TGF-β signaling pathway

Fig.3 The effect of RS4651 on the EMT of AML12 cells with or without SMAD7 silenced (a) R4651 inhibits the protein expression of EMT indicator in AML12 cells through up-regulating SMAD7 (b) RS4651 up-regulates the mRNA expression of SMAD7 in AML12 cells (c) R4651 inhibits the mRNA expression of EMT indicator in AML12 cells through up-regulating SMAD7. ^* P<0.05,^** P<0.01

Fig.4 RS4651 inhibited the proliferation ability of AML12 cells

Fig.5 RS4651 inhibited the migration ability of AML12 cells via SMAD7 The Transwell test showed that RS4651 inhibited the migration of AML12 cells through SMAD7 (original magnification: ×10; scale bars: 100 μm), and the number of migrating cells in each group. ^* P<0.05,^** P<0.01


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