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Int. J. Mol. Sci. 2016, 17(8), 1283; doi:10.3390/ijms17081283

Zeb1 Is a Potential Regulator of Six2 in the Proliferation, Apoptosis and Migration of Metanephric Mesenchyme Cells

1
Division of Molecular Nephrology and The Creative Training Center for Undergraduates, The Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China
2
Department of Laboratory Medicine, The First Hospital of Xi’an, Xi’an 710002, China
3
Undergraduates Class of 2012 Entry, The Fifth Clinical College of Medicine, Chongqing Medical University, Chongqing 400016, China
4
The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Anthony Lemarié
Received: 17 April 2016 / Revised: 21 July 2016 / Accepted: 27 July 2016 / Published: 6 August 2016
(This article belongs to the Collection Programmed Cell Death and Apoptosis)
View Full-Text   |   Download PDF [8270 KB, uploaded 10 August 2016]   |  

Abstract

Nephron progenitor cells surround around the ureteric bud tips (UB) and inductively interact with the UB to originate nephrons, the basic units of renal function. This process is determined by the internal balance between self-renewal and consumption of the nephron progenitor cells, which is depending on the complicated regulation networks. It has been reported that Zeb1 regulates the proliferation of mesenchymal cells in mouse embryos. However, the role of Zeb1 in nephrons generation is not clear, especially in metanephric mesenchyme (MM). Here, we detected cell proliferation, apoptosis and migration in MM cells by EdU assay, flow cytometry assay and wound healing assay, respectively. Meanwhile, Western and RT-PCR were used to measure the expression level of Zeb1 and Six2 in MM cells and developing kidney. Besides, the dual-luciferase assay was conducted to study the molecular relationship between Zeb1 and Six2. We found that knock-down of Zeb1 decreased cell proliferation, migration and promoted cell apoptosis in MM cells and Zeb1 overexpression leaded to the opposite data. Western-blot and RT-PCR results showed that knock-down of Zeb1 decreased the expression of Six2 in MM cells and Zeb1 overexpression contributed to the opposite results. Similarly, Zeb1 promoted Six2 promoter reporter activity in luciferase assays. However, double knock-down of Zeb1 and Six2 did not enhance the apoptosis of MM cells compared with control cells. Nevertheless, double silence of Zeb1 and Six2 repressed cell proliferation. In addition, we also found that Zeb1 and Six2 had an identical pattern in distinct developing phases of embryonic kidney. These results indicated that there may exist a complicated regulation network between Six2 and Zeb1. Together, we demonstrate Zeb1 promotes proliferation and apoptosis and inhibits the migration of MM cells, in association with Six2. View Full-Text
Keywords: Zeb1; Six2; metanephric mesenchyme cells; cell proliferation; cell apoptosis; cell migration Zeb1; Six2; metanephric mesenchyme cells; cell proliferation; cell apoptosis; cell migration
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Gu, Y.; Zhao, Y.; Zhou, Y.; Xie, Y.; Ju, P.; Long, Y.; Liu, J.; Ni, D.; Cao, F.; Lyu, Z.; Mao, Z.; Hao, J.; Li, Y.; Wan, Q.; Kanyomse, Q.; Liu, Y.; Ren, D.; Ning, Y.; Li, X.; Zhou, Q.; Li, B. Zeb1 Is a Potential Regulator of Six2 in the Proliferation, Apoptosis and Migration of Metanephric Mesenchyme Cells. Int. J. Mol. Sci. 2016, 17, 1283.

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