BMP7 plays a critical role in TMEM100-inhibited cell proliferation and apoptosis in mouse metanephric mesenchymal cells in vitro

Ren, Die; Ju, Pan; Liu, Jianing; Ni, Dongsheng; Gu, Yuping; Long, Yaoshui; Zhou, Qin; Xie, Yajun

doi:10.1007/s11626-017-0211-9

Published: 15 December 2017

BMP7 plays a critical role in TMEM100-inhibited cell proliferation and apoptosis in mouse metanephric mesenchymal cells in vitro

Die Ren^1,2,
Pan Ju^1,2,
Jianing Liu^1,2,
Dongsheng Ni^1,2,
Yuping Gu^1,2,
Yaoshui Long^1,2,
Qin Zhou¹ &
Yajun Xie¹

In Vitro Cellular & Developmental Biology - Animal volume 54, pages 111–119 (2018)Cite this article

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Abstract

Kidney mainly arises from the induction of metanephric mesenchymal cells (MM cells) and the ureteric bud (UB). Transmembrane protein-100 (Tmem100) consists of two transmembrane regions with strong temporal and spatial expression characteristics during renal development. However, the function of Tmem100 in mouse embryonic kidney-derived cells remained unclear. We provided qPCR to verify the relationship between Tmem100 and the BMP signal pathway. To clarify the role of Tmem100 in cell proliferation and apoptosis, we carry out EdU incorporation, annexin V- fluorescein isothiocyanate (FITC) apoptosis assay. Here, we find that the knockdown of Tmem100 increases the proliferation and apoptosis of mouse embryonic kidney-derived cells, and this promotion can be inhibited by knockdown of BMP7 at the same time; these results suggest that BMP7 plays a crucial role in Tmem100-regulated cell proliferation and apoptosis. qRT-PCR results further demonstrate that the deficiency of Tmem100 leads to BMP7 upregulation and overexpression could get opposite results. In BMP7-depleted MK3 cells, Tmem100 is highly upregulated and BMPR-II is downregulated. And in BMP7-overexpressed MK3 cells, the expression of Tmem100 is decreased. In BMPR-II-depleted MK3 cells, Tmem100 is downregulated and BMP7 expression remains still. These findings indicate that both BMP7 and BMPR-II can regulate Tmem100 and vice versa, and BMPR-II expression is regulated by BMP7. However, BMP7 has no association with BMPR-II in MK3 cells. Our data demonstrated the significant role of BMP7 in Tmem100-regulated cell proliferation and apoptosis and revealed the complicated regulation network among Tmem100, BMP7, and BMPR-II in mouse embryonic kidney-derived cells.

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Acknowledgements

The authors are grateful to all the fellows in the Division of Molecular Nephrology and the Creative Training Center for undergraduates, the M.O.E. Key Laboratory of Laboratory Medical Diagnostics, the College of Laboratory Medicine, Chongqing Medical University.

Funding

This work is funded by the Natural Science Foundation of China (Grant No. 31271563 and Grant No. 81572076) to Prof. Qin Zhou, M.D. and Ph.D. and granted from the National Basic Research Program of China (No.2011CB944002) to Prof. Qin Zhou, M.D. and Ph.D. This is also supported by the Science and Technology Research Program of Chongqing Municipal Education Commission (Grant No. KJ1702026) by Yajun Xie Ph.D., the Basic Science and Frontier Technology Research Program of Chongqing Science and Technology Commission (Grant No. cstc2017jcyjA0390) by Yajun Xie Ph.D., and the National Science Foundation for Young Scientists of China (Grant No. 31701218) by Yajun Xie Ph.D.

Author information

Die Ren, Pan Ju, and Jianing Liu contributed equally to this work.

Affiliations

Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Laboratory Medical Diagnostics, the College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China
Die Ren, Pan Ju, Jianing Liu, Dongsheng Ni, Yuping Gu, Yaoshui Long, Qin Zhou & Yajun Xie
The College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China
Die Ren, Pan Ju, Jianing Liu, Dongsheng Ni, Yuping Gu & Yaoshui Long

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Die Ren
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Pan Ju
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Jianing Liu
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Dongsheng Ni
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Yuping Gu
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Yaoshui Long
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Qin Zhou
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Yajun Xie
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Correspondence to Qin Zhou or Yajun Xie.

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Editor: Tetsuji Okamoto

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Figure S1

The knockdown of BMPR-II in MK3 cells show the promotion of cell proliferation. Proliferating cells are labeled with EdU (shows as the red positive) and cell nucleus are marked with Dapi (shows as the blue positive) (a.) The BMPR-II-knockdown MK3 cells showed up the promotion of EdU positive cells. (b.) Cell counting of EdU positive cells and total cells are used for analyzing the relative growth rate, n = 3, *: P < 0.05, **: P < 0.01. (PPTX 319 kb)

Figure S2

The knockdown of BMPR-II in MK3 cells show the upregulation of SMAD1/5/8 and SMAD 2/3 mRNA expression. The overexpression of Tmem100 lead to the downregulation of ID1. (a.) qPCR is performed to verified the relative mRNA expression level of BMPR-II, SMAD 1/5/8 and SMAD 2/3 in the knockdown of BMPR-II in MK3 cells, n = 6, *: P < 0.05, ***: P < 0.001. (b.) qPCR is performed to verified the relative mRNA expression level of Tmem100, BMPR-II, ID1 and NANOG in the overexpression of Tmem100 in MK3 cells, n = 3, *: P < 0.05, **: P < 0.01. (PPTX 123 kb)

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Ren, D., Ju, P., Liu, J. et al. BMP7 plays a critical role in TMEM100-inhibited cell proliferation and apoptosis in mouse metanephric mesenchymal cells in vitro. In Vitro Cell.Dev.Biol.-Animal 54, 111–119 (2018). https://doi.org/10.1007/s11626-017-0211-9

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Received: 15 February 2017
Accepted: 17 October 2017
Published: 15 December 2017
Issue Date: February 2018
DOI: https://doi.org/10.1007/s11626-017-0211-9

Keywords

BMP7
Tmem100
Proliferation and apoptosis
Metanephric mesenchymal cells