miR199a-3p regulates P53 by targeting CABLES1 in mouse cardiac c-kit cells to promote proliferation and inhibit apoptosis through a negative feedback loopReport as inadecuate




miR199a-3p regulates P53 by targeting CABLES1 in mouse cardiac c-kit cells to promote proliferation and inhibit apoptosis through a negative feedback loop - Download this document for free, or read online. Document in PDF available to download.

Stem Cell Research and Therapy

, 8:127

First Online: 05 June 2017Received: 27 July 2016Revised: 09 February 2017Accepted: 21 February 2017DOI: 10.1186-s13287-017-0515-4

Cite this article as: Liu, J., Wang, Y., Cui, J. et al. Stem Cell Res Ther 2017 8: 127. doi:10.1186-s13287-017-0515-4

Abstract

BackgroundMicroRNAs miRNAs have emerged as crucial factors that regulate proliferation and apoptosis of cardiac c-kit cells. Although much is known about their role in maintaining cardiac c-kit cell pluripotency, the mechanisms by which they affect cell fate decisions that are an essential part of the repair of heart failure remain poorly understood.

MethodsCardiac c-kit cells were obtained from Balb-c mice and cultured in vitro. Lentiviral vectors of miR199a-3p, its corresponding anti-miRNA, or short hairpin RNA against Cables1 were transfected into cells. The proliferation of cardiac c-kit cells was evaluated using EdU and flow cytometry. Furthermore, we examined cell apoptosis by flow cytometry under treatment with 200nM angiotensin II for 48 h. The levels of miR199a-3p and Cables1 mRNA were measured by quantitative real-time polymerase chain reaction qRT-PCR. Western blot was performed to examine the expression of Cables1 and P53 proteins.

ResultsWe demonstrated a significantly decreased expression of miR199a-3p in heart failure samples compared with healthy donors. Meanwhile, we identified miR199a-3p as a proliferation- and apoptosis-associated regulator impacted through Cdk5 and Abl enzyme substrate 1 CABLES1 targeting, and also attributed their repression to P53 protein expression. We further demonstrated that P53 induced miR199a-3p expression and, in turn, miR199-3p decreased P53 activity.

ConclusionCollectively, our findings uncover one new mechanism by which P53 induced miR199a-3p expression and, in turn, miR199-3p decreased P53 activity. Therefore, miR199a-3p and P53 are coupled through CABLES1 and comprise a novel negative feedback loop that likely contributes to cardiac c-kit cell proliferation and apoptosis.

AbbreviationsAUCArea under the curve

CABLES1Cdk5 and Abl enzyme substrate 1

CDKCyclin-dependent kinase

DMEMDulbecco’s modified Eagle’s medium

EdU5-Ethynyl-2-deoxyuridine

FBSFetal bovine serum

HIF-1aHypoxia-inducible factor-1a

iPSCInduced pluripotent stem cell

MEFMouse embryonic fibroblast

miRNAMicroRNA

PBSPhosphate-buffered saline

PIPropidium iodide

qRT-PCRQuantitative real-time polymerase chain reaction

RAASRenin–angiotensin–aldosterone system

ROCReceiver-operating characteristic

SNSSympathetic nervous system

TUTransducing units

UTRUntranslated region





Author: Jingjin Liu - Yongshun Wang - Jinjin Cui - Meng Sun - Zhongyue Pu - Chao Wang - Wenjuan Du - Xinxin Liu - Jian Wu - Jingb

Source: https://link.springer.com/



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