Abstract
Contact inhibition and its disruption of vascular smooth muscle cells (VSMCs) are important cellular events in vascular diseases. But the underlying molecular mechanisms are unclear. In this study we investigated the roles of microRNAs (miRNAs) in the contact inhibition and its disruption of VSMCs and the molecular mechanisms involved. Rat VSMCs were seeded at 30% or 90% confluence. MiRNA expression profiles in contact-inhibited confluent VSMCs (90% confluence) and non-contact-inhibited low-density VSMCs (30% confluence) were determined. We found that multiple miRNAs were differentially expressed between the two groups. Among them, miR-145 was significantly increased in contact-inhibited VSMCs. Serum could disrupt the contact inhibition as shown by the elicited proliferation of confluent VSMCs. The contact inhibition disruption accompanied with a down-regulation of miR-145. Serum-induced contact inhibition disruption of VSMCs was blocked by overexpression of miR-145. Moreover, downregulation of miR-145 was sufficient to disrupt the contact inhibition of VSMCs. The downregulation of miR-145 in serum-induced contact inhibition disruption was related to the activation PI3-kinase/Akt pathway, which was blocked by the PI3-kinase inhibitor LY294002. KLF5, a target gene of miR-145, was identified to be involved in miR-145-mediated effect on VSMC contact inhibition disruption, as it could be inhibited by knockdown of KLF5. In summary, our results show that multiple miRNAs are differentially expressed in contact-inhibited VSMCs and in non-contact-inhibited VSMCs. Among them, miR-145 is a critical gene in contact inhibition and its disruption of VSMCs. PI3-kinase/Akt/miR-145/KLF5 is a critical signaling pathway in serum-induced contact inhibition disruption. Targeting of miRNAs related to the contact inhibition of VSMCs may represent a novel therapeutic approach for vascular diseases.
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Acknowledgements
This work was supported by the National Institutes of Health (No 1R01HL130052) to Chun-xiang ZHANG. This work was also supported by the National Natural Science Foundation of China (No 81670398 and 91639102), the Natural Foundation of Shandong Province (No ZR2013CL003), Qing-dao LING Jun-ren CAI Fund (No #13-CX-3) and the funding of Taishan Scholars of Shandong Province to Binzhou Medical University.
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Supplementary information is available at the website of Acta Pharmacologica Sinica.
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Supplementary Figure S1
Thirty-five miRNAs that were highly expressed in VSMCs and over 30% difference in their expression signals between low density and high density cells. (JPG 64 kb)
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Sun, Yy., Qin, Ss., Cheng, Yh. et al. MicroRNA expression profile and functional analysis reveal their roles in contact inhibition and its disruption switch of rat vascular smooth muscle cells. Acta Pharmacol Sin 39, 885–892 (2018). https://doi.org/10.1038/aps.2018.6
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DOI: https://doi.org/10.1038/aps.2018.6
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