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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2021, Vol. 41 Issue (2/3): 1-6    DOI: 10.13523/j.cb.2011023
    
PHP14 Plays a Role in Epithelial-Mesenchymal Transition of AML-12Cell Through Interaction with Vimentin
XU An-jian1,**(),LI Yan-meng1,WU Shan-na2,ZHANG Bei1,YAO Jing-yi1
1 Experimental Center, Beijing Friendship Hospital, Beijing 100875, China
2 Clinical Laboratory Center, Beijing Friendship Hospital, Beijing 100875, China
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Abstract  

Objective: To investigate the role of PHP14 in epithelial-mesenchymal transition and its possible mechanism. Methods: A cell model was established to study epithelial-mesenchymal transition in mouse hepatocytes AML-12. Overexpression of PHP14 in AML-12. Morphological observation was carried out by light microscope. The expression of related markers in epithelial-mesenchymal transition of AML-12 was detected by real-time fluorescent quantitative PCR and Western blot. The interaction between PHP14 and Vimentin was detected by immunoprecipitation. Results: Mouse hepatocyte AML-12 can be used as a good cell model to study epithelial-mesenchymal transition. Overexpression of PHP14 in AML-12 can promote epithelial-mesenchymal transition and enhance the motility of AML-12 cells. Overexpression of PHP14 upregulated Vimentin, but had no effect on the expression of other epithelial-mesenchymal transition related markers. Further studies showed that PHP14 could interact with Vimentin. Conclusion: PHP14 can stabilize Vimentin protein level and participate in and regulate epithelial-mesenchymal transition, by interacting with Vimentin.

Key words： PHP14      Epithelial-mesenchymal transition      AML-12 Vimentin      Cell migration     
Received: 11 November 2020      Published: 08 April 2021
ZTFLH:  Q819  
Corresponding Authors: An-jian XU     E-mail: xuanjian@ccmu.edu.com
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An-jian XU
Yan-meng LI
Shan-na WU
Bei ZHANG
Jing-yi YAO
Cite this article:

XU An-jian,LI Yan-meng,WU Shan-na,ZHANG Bei,YAO Jing-yi. PHP14 Plays a Role in Epithelial-Mesenchymal Transition of AML-12Cell Through Interaction with Vimentin. China Biotechnology, 2021, 41(2/3): 1-6.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2011023     OR     https://manu60.magtech.com.cn/biotech/Y2021/V41/I2/3/1

Fig.1 TGF-β can induce epithelial mesenchymal transition in mouse hepatocyte line AML-12 (a) Cell morphology change of AML-12 cells after TGF-β treatment (b)The mRNA expression of EMT marker in AML-12 cells after TGF-β treatment. Means without a common superscript differ significantly (P<0.05) (c) The protein expression of EMT marker in AML-12 cells after TGF-β treatment
Fig.2 PHP14 can promote the epithelial mesenchymal transition and migration of AML-12 cells in vitro (a)The protein expression of PHP14 in AML-12 cells after TGF-β treatment (b)The protein expression of PHP14 in control and PHP14 overexpression AML-12 cells (c)The cell morphology change of AML-12 cells after PHP14 overexpression (d)The migration of AML-12 cells after PHP14 overexpression Means without a common superscript differ significantly (P<0.05)
Fig.3 Overexpression of PHP14 in AML-12 cells can increase the protein expression of vimentin induced by TGF-β (a)The mRNA expression of EMT marker in AML-12 cells after PHP14 overexpression Means without a common superscript differ significantly (P<0.05) (b) The protein expression of EMT marker in AML-12 cells after PHP14 overexpression
Fig.4 PHP14 interacts with vimentin The interaction of PHP14 with vimentin was detected by immune-coprecipitation
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