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Downregulation of Kcnq1ot1 attenuates β-cell proliferation and insulin secretion via the miR-15b-5p/Ccnd1 and Ccnd2 axis

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Abstract

Aim

To examine the effect of lncRNA Kcnq1ot1 on pancreatic β cells in the development of diabetes.

Methods

The expression levels of Kcnq1ot1 were detected in the islets of diabetes mouse models and the serum of patients with type 2 diabetes by qRT-PCR. CCK8, Ki67 staining, immunohistochemical analyses, glucose-stimulated insulin secretion and intraperitoneal glucose tolerance test were performed to detect the effect of Kcnq1ot1 on β-cell proliferation and insulin secretion in vitro and in vivo. The relationship between Kcnq1ot1 and miR-15b-5p was predicted by bioinformatics prediction, which was confirmed by luciferase reporter assay.

Results

Kcnq1ot1 was more abundant in the pancreas. The expression of Kcnq1ot1 was decreased in the islets of db/db mice and diet-induced obese mice and in the serum of patients with type 2 diabetes. Silencing Kcnq1ot1 inhibited the β-cell proliferation concomitant with a reduction in the levels of Ccnd1 and Ccnd2. Insulin synthesis and secretion were impaired, along with the decreased expression of Ins1, Ins2, and insulin-related transcription factors. Moreover, Kcnq1ot1 knockdown in vivo reduced glucose tolerance and decreased insulin secretion, consistent with the reduction in the relative islet area and Ki67-positive β-cells detected by immunochemistry and immunofluorescence staining, respectively. Mechanistically, Kcnq1ot1 directly targeted miR-15b-5p which regulated β-cell proliferation and insulin secretion through Ccnd1 and Ccnd2. Notably, the suppression of miR-15b-5p attenuated the inhibition of Min6 proliferation and insulin production induced by Kcnq1ot1 knockdown.

Conclusion

Kcnq1ot1 regulated β-cell proliferation and insulin secretion via the miR-15b-5p/Ccnd1 and Ccnd2 axis, which is worthy of further investigation considering its potential in diabetes treatment.

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Acknowledgements

We thank Dr. Dalong Song (Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences) for his excellent technical assistance. This work was supported by National Natural Science Foundation of China (grants number: 81800682 to Yanli Li); the Medical and Health Project of Guangzhou (grants number: 20201A011079 to Yanli Li); Guangzhou Science and Technology Project (grants number: 202102010175 to Wen Xu).

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Author notes

  1. Yanli Li and Yalan Chen have contributed equally to this work.

Authors and Affiliations

  1. Department of Endocrinology, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510260, China

    Yanli Li, Xiaoyi Deng, Zhishan Chen, Huiyu Ye & Wangen Li

  2. Key Laboratory of Diabetology of Guangdong Province, Department of Endocrinology and Metabolism, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510630, China

    Yalan Chen, Ziyu Liu, Beisi Lin, Danrui Chen, Yanna Su & Wen Xu

  3. Department of Clinical Laboratory, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, China

    Qiwen Xiao

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Contributions

Yanli Li conceived and designed the study, and wrote the manuscript. Yalan Chen, Ziyu Liu, Beisi Lin, Yanna Su and Danrui Chen performed the experiments and analyzed the data. Xiaoyi Deng, Qiwen Xiao, Zhishan Chen, and Huiyu Ye collected specimens and analyzed the data. Wangen Li and Wen Xu contributed to discussion and revision of the manuscript. All the authors agreed on the final manuscript.

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Correspondence to Wangen Li or Wen Xu.

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All animal experiments were approved by the Institutional Animal Care and Use Committee of Sun Yat-Sen University. The studies involving human participants were reviewed and approved by the Ethics Committee of the Second Affiliated Hospital of Guangzhou University.

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Li, Y., Chen, Y., Liu, Z. et al. Downregulation of Kcnq1ot1 attenuates β-cell proliferation and insulin secretion via the miR-15b-5p/Ccnd1 and Ccnd2 axis. Acta Diabetol 59, 885–899 (2022). https://doi.org/10.1007/s00592-022-01871-6

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