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      Shikonin Inhibits the Proliferation of Human Breast Cancer Cells by Reducing Tumor-Derived Exosomes

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          Abstract

          Shikonin is a naphthoquinone isolated from the traditional Chinese medicine Lithospermum. It has been used in the treatment of various tumors. However, the effects of shikonin on such diseases have not been fully elucidated. In the present study, we detected the exosome release of a breast cancer cell line (MCF-7) with shikonin treatment and found a positive relationship between the level of secreted exosomes and cell proliferation. We next analyzed miRNA profiles in MCF-7 cells and exosomes and found that some miRNAs are specifically sorted and abundant in exosomes. Knockdown of the most abundant miRNAs in exosomes and the MCF-7 proliferation assay showed that miR-128 in exosomes negatively regulates the level of Bax in MCF-7 recipient cells and inhibits cell proliferation. These results show that shikonin inhibits the proliferation of MCF-7 cells through reducing tumor-derived exosomal miR-128. The current study suggests that shikonin suppresses MCF-7 growth by the inhibition of exosome release.

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          Pharmacological properties of shikonin - a review of literature since 2002.

          Rosa Giner, Francisco J. O. Rios, Isabel Andújar … (2013)
          The naphthoquinone shikonin is the main active principle of Zicao, a traditional Chinese herbal medicine made from the dried root of Lithospermum erythrorhizon. Studies carried out over the past 30 years have provided a scientific basis for the use of Zicao which has been long employed in folk medicine to treat a variety of inflammatory and infectious diseases. In particular, shikonin has been shown to possess many diverse properties, including antioxidant, anti-inflammatory, antithrombotic, antimicrobial, and wound healing effects. The fact that shikonin shows so many beneficial properties has increased the interest in this molecule dramatically, especially in the past few years. The aim of this review is to provide an update of the new data published on shikonin, whose wide spectrum of pharmacological effects as well as pharmacokinetic properties and toxicity make it a highly interesting target molecule. Georg Thieme Verlag KG Stuttgart · New York.
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            Exosome-mediated transfer of miR-222 is sufficient to increase tumor malignancy in melanoma

            Federica Felicetti, Alessandra De Feo, Carolina Coscia … (2016)
            Background Growing evidence is showing that metastatic cell populations are able to transfer their characteristics to less malignant cells. Exosomes (EXOs) are membrane vesicles of endocytic origin able to convey their cargo of mRNAs, microRNAs (miRs), proteins and lipids from donors to proximal as well as distant acceptor cells. Our previous results indicated that miR-221&222 are key factors for melanoma development and dissemination. The aim of this study was to verify whether the tumorigenic properties associated with miR-222 overexpression can be also propagated by miR-222-containing EXOs. Methods EXOs were isolated by UltraCentrifugation or Exoquick-TC® methods. Preparations of melanoma-derived vesicles were characterized by using the Nanosight™ technology and the expression of exosome markers analyzed by western blot. The expression levels of endogenous and exosomal miRNAs were examined by real time PCR. Confocal microscopy was used to evaluate transfer and uptake of microvesicles from donor to recipient cells. The functional significance of exosomal miR-222 was estimated by analyzing the vessel-like process formation, as well as cell cycle rates, invasive and chemotactic capabilities. Results Besides microvesicle marker characterization, we evidenced that miR-222 exosomal expression mostly reflected its abundance in the cells of origin, correctly paralleled by repression of its target genes, such as p27Kip1, and induction of the PI3K/AKT pathway, thus confirming its functional implication in cancer. The possible differential significance of PI3K/AKT blockade was assessed by using the BKM120 inhibitor in miR-222-transduced cell lines. In addition, in vitro cultures showed that vesicles released by miR-222-overexpressing cells were able to transfer miR-222-dependent malignancy when taken-up by recipient primary melanomas. Results were confirmed by antagomiR-221&222 treatments and by functional observations after internalization of EXOs devoid of these miRs. Conclusion All together these data, besides generally confirming the role of miR-222 in melanoma tumorigenesis, supported its responsibility in the exosome-associated melanoma properties, thus further indicating this miR as potential diagnostic and prognostic biomarker and its abrogation as a future therapeutic option. Electronic supplementary material The online version of this article (doi:10.1186/s12967-016-0811-2) contains supplementary material, which is available to authorized users.
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              Tumor-cell-derived microvesicles as carriers of molecular information in cancer.

              Pierre Hainaut, A M V C Martins, Ana Dias (2012)
              Exosomes and microvesicles are secreted particles of 30-200  nm in diameter, delimited by a lipid bilayer and containing a wide range of membrane-bound or free proteins and nucleic acids (in particular mRNA and miRNA). Here, we review the properties of tumor-cell-derived microvesicles as carriers of molecular information in relation to cancer progression and promotion of metastasis. Microvesicles from tumor cells operate as signaling platforms that diffuse in the extracellular space to target cells in the microenvironment, modulating the interactions of tumor cells with stromal, inflammatory, dendritic, immune or vascular cells and priming the formation of the metastatic niche. Because of their stability, exosomes and microvesicles can be retrieved in bodily fluids as biomarkers for cancer detection and monitoring. They offer a range of molecular targets for controlling cell-cell interactions during invasion and metastasis.
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                Author and article information

                Contributors
                Thomas J. Schmidt: Role: Academic Editor
                Journal
                Journal ID (nlm-ta): Molecules
                Journal ID (iso-abbrev): Molecules
                Journal ID (publisher-id): molecules
                Title: Molecules
                Publisher: MDPI
                ISSN (Electronic): 1420-3049
                Publication date (Electronic): 16 June 2016
                Publication date Collection: June 2016
                Volume: 21
                Issue: 6
                Electronic Location Identifier: 777
                Affiliations
                State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Engineering Research Center for MicroRNA Biology and Biotechnology, Nanjing Advanced Institute for Life Sciences, School of Life Sciences, Nanjing University, Nanjing 210093, Jiangsu, China; nju_weiyao@ 123456163.com (Y.W.); 15261891180@ 123456163.com (M.L.); shufangnice@ 123456163.com (S.C.); biowonder@ 123456126.com (D.W.)
                Author notes
                [* ]Correspondence: cyzhang@ 123456nju.edu.cn (C.-Y.Z.); kzen@ 123456nju.edu.cn (K.Z.); bixuehan2005@ 123456163.com (L.L.); Tel.: +86-25-8968-0249 (L.L.)
                [†]

                These authors contributed equally to this work.

                Article
                Publisher ID: molecules-21-00777
                DOI: 10.3390/molecules21060777
                PMC ID: 6274101
                PubMed ID: 27322220
                SO-VID: 50a05cb9-de7a-4b5e-9e56-2b944741f9d1
                Copyright © © 2016 by the authors.
                License:

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                Date received : 11 April 2016
                Date accepted : 07 June 2016
                Categories
                Subject: Article

                Keywords: breast cancer,proliferation,shikonin,exosome,mir-128,bax
                Data availability:
                Keywords: breast cancer, proliferation, shikonin, exosome, mir-128, bax

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