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miR-33-3p Regulates PC12 Cell Proliferation and Differentiation In Vitro by Targeting Slc29a1

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Abstract

MicroRNA-33-3p (miR-33-3p) has been widely investigated for its roles in lipid metabolism and mitochondrial function; however, there are few studies on miR-33-3p in the context of neurological diseases. In this study, we investigated the functional role of miR-33-3p in rat pheochromocytoma PC12 cells. A miR-33-3p mimic was transduced into PC12 cells, and its effects on proliferation, apoptosis, and differentiation were studied using the MTS assay, EdU labeling, flow cytometry, qRT-PCR, western blot, ELISA, and immunofluorescence. We found that miR-33-3p significantly suppressed PC12 cell proliferation, but had no effect on apoptosis. Furthermore, miR-33-3p promoted the differentiation of PC12 cells into Tuj1-positive and choline acetyltransferase-positive neuron-like cells. Mechanistically, miR-33-3p repressed the expression of Slc29a1 in PC12 cells. Importantly, knocking down Slc29a1 in PC12 cells inhibited proliferation and induced differentiation into neuron-like cells. In conclusion, this study showed that miR-33-3p regulated Slc29a1, which in turn controlled the proliferation and differentiation of PC12 cells. Thus, we hypothesize that the miR-33-3p/Slc29a1 axis could be a promising therapeutic target for recovering neurons and the cholinergic nervous system.

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Acknowledgements

We gratefully acknowledge the Graduate Scientific Research Innovation Program of Jiangsu Province, Natural Science Foundation of Jiangsu Province and Nantong University, and a Project Funded by the Priority Academic Program Development (PAPD) of Jiangsu Higher Education institutions for the research grants. We also acknowledge the editor and reviewers for their helpful comments on this paper.

Funding

Contract grant sponsor: Graduate Scientific Research Innovation Program of Jiangsu Province; Contract Grant Number: KYCX19 2066. Contract grant sponsor: Natural Science Foundation of Jiangsu Province; Contract Grant Number: BK20170447. Contract grant sponsor: a Project Funded by the Priority Academic Program Development (PAPD) of Jiangsu Higher Education institutions. Contract Grant Number: 03081023.

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Author notes

  1. Bo-Quan Shan and Wen Li have contributed equally to this work.

Authors and Affiliations

  1. Department of Human Anatomy, Institute of Neurobiology, Nantong University, No.19 Qixiu Road, No.3 Building of Qixiu Campus, Nantong, 226001, Jiangsu, China

    Bo-Quan Shan, Wen Li, Hui He, He-Yan Zhao, Mei-Ling Tian, Xiang Cheng, Jian-Bing Qin & Guo-Hua Jin

  2. Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, No.19 Qixiu Road, No.3 Building of Qixiu Campus, Nantong, 226001, Jiangsu, China

    Bo-Quan Shan, Wen Li, Hui He, He-Yan Zhao, Mei-Ling Tian, Xiang Cheng, Jian-Bing Qin & Guo-Hua Jin

  3. Co-Innovation Center of Neuroregeneration, Nantong University, No.19 Qixiu Road, No.3 Building of Qixiu Campus, Nantong, 226001, Jiangsu, China

    Bo-Quan Shan, Wen Li, Hui He, He-Yan Zhao, Mei-Ling Tian, Xiang Cheng, Jian-Bing Qin & Guo-Hua Jin

  4. Department of Anatomy and Neurobiology, Jiangsu Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Collaborative Innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu, China

    Jian-Bing Qin & Guo-Hua Jin

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  1. Bo-Quan Shan
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  2. Wen Li
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Contributions

BS, and WL conceived the experiments. BS performed the experimenets with help from HZ, MT, XC, JQ. BS and GJ analyzed the data. BS, JQ, and GJ wrote the manuscript. All authors read, revised, and approved the manuscript.

Corresponding authors

Correspondence to Jian-Bing Qin or Guo-Hua Jin.

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Fig. S1 Interference efficiency of Slc29a1 was detected by qRT-PCR and Western blot.

Supplementary file2 (PDF 2236 KB)

Fig. S2 Effects of miR-33-3p-Slc29a1 on NSCs differentiation. a,b The concentration of ACh and ChAT level were detected with ELISA assay. c,d Immunocytochemical analyse was applied to detected he percentage of Tuj1 (red) and ChA T(red) immunopositive cells. Nuclei are visualized with Hoechst (blue), and analyzed by fluorescent microscopy. Scale bar, 100 μm. Data are expressed as the mean ± SD; n = 3. **P<0.01, ***P<0.001.

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Shan, BQ., Li, W., He, H. et al. miR-33-3p Regulates PC12 Cell Proliferation and Differentiation In Vitro by Targeting Slc29a1. Neurochem Res 46, 2403–2414 (2021). https://doi.org/10.1007/s11064-021-03377-z

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  • DOI: https://doi.org/10.1007/s11064-021-03377-z

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