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hsa-miR-4485 regulates mitochondrial functions and inhibits the tumorigenicity of breast cancer cells

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Abstract

The modulation of mitochondrial functions is important for maintaining cellular homeostasis. Mitochondria essentially depend on the import of RNAs and proteins encoded by the nuclear genome. MicroRNAs encoded in the nucleus can translocate to mitochondria and target the genome, affecting mitochondrial function. Here, we analyzed the role of miR-4485 in the regulation of mitochondrial functions. We showed that miR-4485 translocated to mitochondria where its levels varied in response to different stress conditions. A direct binding of miR-4485 to mitochondrial 16S rRNA was demonstrated. MiR-4485 regulated the processing of pre-rRNA at the 16S rRNA-ND1 junction and the translation of downstream transcripts. MiR-4485 modulated mitochondrial complex I activity, the production of ATP, ROS levels, caspase-3/7 activation, and apoptosis. Transfection of a miR-4485 mimic downregulated the expression of regulatory glycolytic pathway genes and reduced the clonogenic ability of breast cancer cells. Ectopic expression of miR-4485 in MDA-MB-231 breast carcinoma cells decreased the tumorigenicity in a nude mouse xenograft model. Furthermore, levels of both precursor and mature miR-4485 are decreased in tumor tissue of breast cancer patients. We conclude that the mitochondria-targeted miR-4485 may act as a tumor suppressor in breast carcinoma cells by negatively regulating mitochondrial RNA processing and mitochondrial functions.

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Acknowledgements

This work is being supported by EMR/2014/000219, DST, Government of India to RS. This work is lead obtained from previous DBT grant BT/PR12906/AGR/36/638/2009, DBT, Government of India. A.V.L. and P.M.C. were supported by the grant 14-50-00060 from the Russian Science Foundation. The authors acknowledge the instrumentation facility by DBT MSUB ILSPARE. We thank Prof. Narry Kim, Institute for Basic Science and Seoul National University, Korea for giving us plasmids of pCK-Flag-Dicer as a gift. L.S., A.S., and K.S. received their Senior Research fellowship from University Grants Commission (UGC), Government of India. K.B. received her Senior Research fellowship from the Council of Scientific and Industrial Research (CSIR), Government of India. Dr. Rochika Singh received her fellowship as DST Young Scientist from Department of Science and Technology (DST), Government of India. This work consists of the PhD thesis of L.S.

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Authors and Affiliations

  1. Department of Biochemistry, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat, 390002, India

    Lakshmi Sripada, Kritarth Singh, Paresh Prajapati, Khyati Bhatelia, Milton Roy & Rajesh Singh

  2. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia

    Anastasiya V. Lipatova & Peter M. Chumakov

  3. Department of Endocrinology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, Uttar Pradesh, 226014, India

    Aru Singh & Madan M. Godbole

  4. Center for Translational Medicine, Temple University, Philadelphia, PA, 19140, USA

    Dhanendra Tomar

  5. Department of Cell Biology, School of Biological Sciences and Biotechnology, Indian Institute of Advanced Research, Koba Institutional Area, Gandhinagar, Gujarat, 382007, India

    Rochika Singh

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  1. Lakshmi Sripada
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Sripada, L., Singh, K., Lipatova, A.V. et al. hsa-miR-4485 regulates mitochondrial functions and inhibits the tumorigenicity of breast cancer cells. J Mol Med 95, 641–651 (2017). https://doi.org/10.1007/s00109-017-1517-5

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  • DOI: https://doi.org/10.1007/s00109-017-1517-5

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