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Identification of miR-9 as a negative factor of insulin secretion from beta cells

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Abstract

MicroRNA is a novel class of small noncoding RNA that has been implicated in a variety of physiological and pathological processes, including glucose homeostasis and diabetes mellitus. So far, a few studies have reported that miRNAs may be an important regulator in glucose-stimulated insulin secretion (GSIS) pathway. However, the role of miRNAs in this process remains unclear. The levels of miRNAs in mouse islets and MIN6 cells were determined by quantitative RT-PCR. Concentration of insulin was determined by ELISA, and the expression of the target protein was determined with western blot assay. The overexpression and downregulation of miRNAs in MIN6 were conducted using cell transfection methods. And luciferase assay was used to measure the direct interaction between miRNAs and target messenger RNAs 3′UTR. miR-9 was screened out for it was downregulated under the effects of short-term high glucose, while long-term high glucose relatively increased miR-9 expression. The Stxbp1 expression was decreased with the overexpression of miR-9 in MIN6 cells and increased when miR-9 was downregulated. Moreover, it was verified by luciferase assay that miR-9 regulated Stxbp1 gene expression by directly targeting Stxbp1 messenger RNA 3′UTR. This study suggests that the pathway consisting of miR-9 and Stxbp1 plays a key role in β-cell function, thus contributing to the network of miRNA-insulin secretion and offering a new candidate for diabetes therapy.

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Funding

This work was supported by grants from the National Natural Science Foundation of China (No. 81602158). The funders had no role in study design; in the collection, analysis, and interpretation of data; in the writing of the report; and in the decision to submit this article for publication.

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Authors and Affiliations

  1. Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin’s Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin, 300060, China

    Dongzhi Hu, Yi Wang, Haiyang Zhang & Dalu Kong

Authors
  1. Dongzhi Hu
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  2. Yi Wang
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  3. Haiyang Zhang
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  4. Dalu Kong
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Contributions

Dongzhi Hu and Yi Wang performed most of the experiments, analyzed data, and wrote the manuscript. Dalu Kong and Haiyang Zhang designed the experiments and edited the manuscript. Dalu Kong is the guarantor of this work, has full access to all of the data in the study, and takes responsibility for the integrity of the data and the accuracy of the data analysis.

Corresponding authors

Correspondence to Haiyang Zhang or Dalu Kong.

Ethics declarations

The study was performed in accordance with protocols approved by the Institutional Animal Care and Research Advisory Committee of Tianjin Medical University Cancer Institute and Hospital.

Conflict of interest

The authors declare that there is no conflict of interest.

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Hu, D., Wang, Y., Zhang, H. et al. Identification of miR-9 as a negative factor of insulin secretion from beta cells. J Physiol Biochem 74, 291–299 (2018). https://doi.org/10.1007/s13105-018-0615-3

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  • DOI: https://doi.org/10.1007/s13105-018-0615-3

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