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circ-CCND1 regulates the CCND1/P53/P21 pathway through sponging miR-138-5p in valve interstitial cells to aggravate aortic valve calcification

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A Correction to this article was published on 19 July 2022

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Abstract

To discuss the effect and mechanism of circular-CCND1 (circ-CCND1) on the regulation of calcified aortic valve disease (CAVD). Differentially expressed circRNAs were screened through the GSE155119 data set and biological prediction. Subsequently, the miR-138-5p, CCND1, and circ-CCND1 expression were detected in the non-calcified and calcified aortic valve. Then Pearson correlation analysis was performed to analyze the correlation between the above expression, and dual luciferase and RNA-pull down assays for verifying the target relationship. Porcine aortic valve interstitial cells (AVICs) were isolated and transfected with pcDNA-circ-CCND1, miR-138-5p inhibitor, and miR-138-5p mimics. The alkaline phosphatase (ALP) activity was quantitatively analyzed by ALP staining, and alizarin-red staining was to check the calcium nodules formation. Finally, Western blot was applied to detect the expression of proteins associated with osteogenic differentiation (Runx2, Osterix, OPN) and CCND1/P53/P21 pathway proteins. Circ-CCND1 was highly expressed in calcific aortic valves. After inhibiting circ-CCND1 expression, a significant reduction was shown in ALP activity, the degree of ossification and the formation of calcium nodules in AVICs, and osteogenic differentiation-related protein expression and CCND1/P53/P21 pathway protein expression. By contrast, inhibition of miR-138-5p and circ-CCND1 together promoted the calcification of AVICs and expression of CCND1/P53/P21 pathway proteins. P53 inhibitor (PFT-α) could significantly reduce activation of CCND1/P53/P21 pathway protein expression by circ-CCND1 overexpression. However, P53 activator (Nutlin-3) significantly restored the suppression of the above pathway-related protein expression by downregulation of circ-CCND1. Circ-CCND1 sponges miR-138-5p to regulate CCND1 expression, thereby promoting the calcification of AVICs.

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    Authors and Affiliations

    1. Department of Cardiac Surgery, The First Affiliated Hospital of Xinjiang Medical University, Xinshi District, 137 Liyushan South Road, Urumqi, 830054, Xinjiang, China

      Fei Yan, Qiang Huo, Weimin Zhang,  Daniyaer·Dilimulati & Lin Shi

    2. Department of Cardiology Surgery, The First Affiliated Hospital of Xinjiang Medical University, Xinshi District, 137 Liyushan South Road, Urumqi, 830054, Xinjiang, China

      Xiang Xie & Tingting Wu

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    1. Fei Yan
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    2. Xiang Xie
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    3. Qiang Huo
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    Contributions

    YF contributed to conception and design of the study. XX, QH and WZ performed the statistical analysis. YF wrote the first draft of the manuscript. XX, QH, WZ, TW, DD, and LS wrote sections of the manuscript. All authors contributed to manuscript revision, read, and approved the submitted version. The authors declare that all data were generated in-house and that no paper mill was used.

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    Correspondence to Fei Yan.

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    This study was approved by the Medical Ethics Committee of The First Affiliated Hospital of Xinjiang Medical University (K202107-09).

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    Key points

    1)Circ-CCND1 regulates the expression of CCND1 through sponging miR-138-5p and therefore promotes the calcification of AVIC cells.

    2)Circ-CCND1 is highly expressed in calcified aortic valve leaflets.

    3)Circ-CCND1 can be used as a diagnostic marker for CAVD.

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    Yan, F., Xie, X., Huo, Q. et al. circ-CCND1 regulates the CCND1/P53/P21 pathway through sponging miR-138-5p in valve interstitial cells to aggravate aortic valve calcification. J Physiol Biochem 78, 845–854 (2022). https://doi.org/10.1007/s13105-022-00907-3

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    • DOI: https://doi.org/10.1007/s13105-022-00907-3

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