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      Mxi1-0 regulates the growth of human umbilical vein endothelial cells through extracellular signal-regulated kinase 1/2 (ERK1/2) and interleukin-8 (IL-8)-dependent pathways

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          Abstract

          Mxi1 plays an important role in the regulation of cell proliferation. Mxi1-0, a Mxi1 isoform, has a different N-terminal amino acid sequence, intracellular location and expression profile from Mxi1. However, the precise role of Mxi1-0 in cell proliferation and the molecular mechanism underlying its function remain poorly understood. Here, we showed that Mxi1-0 suppression decreased the proliferation of human umbilical vein endothelial cells (HUVECs) along with cell accumulation in the G2/M phase. Mxi1-0 suppression also significantly decreased the expression and secretion of interleukin (IL-8). Neutralizing IL-8 in conditioned medium (CM) from Mxi1-0-overexpressed HUVECs significantly eliminated CM-induced proliferation of HUVECs. In addition, Mxi1-0 suppression significantly decreased the activity of MAP kinase ERK1/2. Treatment of HUVECs with U0126, an ERK1/2 signaling inhibitor, attenuated autocrine production of IL-8 induced by Mxi1-0 overexpression. On the other hand, Mxi1-0 overexpression-induced IL-8 increased the level of phosphorylated ERK1/2 in HUVECs, and such increasing was diminished in cells incubated with CM, which neutralized with anti-IL-8 antibody. Taken together, our results suggest that Mxi1-0 regulates the growth of HUVECs via the IL-8 and ERK1/2 pathways, which apparently reciprocally activate each other.

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          Growth of triple-negative breast cancer cells relies upon coordinate autocrine expression of the proinflammatory cytokines IL-6 and IL-8.

          Triple-negative breast cancers (TNBC) are aggressive with no effective targeted therapies. A combined database analysis identified 32 inflammation-related genes differentially expressed in TNBCs and 10 proved critical for anchorage-independent growth. In TNBC cells, an LPA-LPAR2-EZH2 NF-κB signaling cascade was essential for expression of interleukin (IL)-6, IL-8, and CXCL1. Concurrent inhibition of IL-6 and IL-8 expression dramatically inhibited colony formation and cell survival in vitro and stanched tumor engraftment and growth in vivo. A Cox multivariable analysis of patient specimens revealed that IL-6 and IL-8 expression predicted patient survival times. Together these findings offer a rationale for dual inhibition of IL-6/IL-8 signaling as a therapeutic strategy to improve outcomes for patients with TNBCs. ©2013 AACR.
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            Cdc25 phosphatases are required for timely assembly of CDK1-cyclin B at the G2/M transition.

            Progression through mitosis requires the coordinated regulation of Cdk1 kinase activity. Activation of Cdk1 is a multistep process comprising binding of Cdk1 to cyclin B, relocation of cyclin-kinase complexes to the nucleus, activating phosphorylation of Cdk1 on Thr(161) by the Cdk-activating kinase (CAK; Cdk7 in metazoans), and removal of inhibitory Thr(14) and Tyr(15) phosphorylations. This dephosphorylation is catalyzed by the dual specific Cdc25 phosphatases, which occur in three isoforms in mammalian cells, Cdc25A, -B, and -C. We find that expression of Cdc25A leads to an accelerated G(2)/M phase transition. In Cdc25A-overexpressing cells, Cdk1 exhibits high kinase activity despite being phosphorylated on Tyr(15). In addition, Tyr(15)-phosphorylated Cdk1 binds more cyclin B in Cdc25A-overexpressing cells compared with control cells. Consistent with this observation, we demonstrate that in human transformed cells, Cdc25A and Cdc25B, but not Cdc25C phosphatases have an effect on timing and efficiency of cyclin-kinase complex formation. Overexpression of Cdc25A or Cdc25B promotes earlier assembly and activation of Cdk1-cyclin B complexes, whereas repression of these phosphatases by short hairpin RNA has a reverse effect, leading to a substantial decrease in amounts of cyclin B-bound Cdk1 in G(2) and mitosis. Importantly, we find that Cdc25A overexpression leads to an activation of Cdk7 and increase in Thr(161) phosphorylation of Cdk1. In conclusion, our data suggest that complex assembly and dephosphorylation of Cdk1 at G(2)/M is tightly coupled and regulated by Cdc25 phosphatases.
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              Bone marrow-derived mesenchymal stem cell-secreted IL-8 promotes the angiogenesis and growth of colorectal cancer

              Mesenchymal stem cells (MSCs) have recently been shown to home to tumors and contribute to the formation of the tumor-associated stroma. In addition, MSCs can secrete paracrine factors to facilitate tumor progression. However, the involvement of MSC-derived cytokines in colorectal cancer (CRC) angiogenesis and growth has not been clearly addressed. In this study, we report that interleukin-8 (IL-8) was the most highly upregulated pro-angiogenic factor in MSCs co-cultured with CRC cells and was expressed at substantially higher levels in MSCs than CRC cells. To evaluate the effect of MSC-derived IL-8 on CRC angiogenesis and growth, we used MSCs that expressed small hairpin (interfering) RNAs (shRNA) targeting IL-8 (shIL-8-MSCs). We found that MSC-secreted IL-8 promoted human umbilical vein endothelial cell (HUVEC) proliferation and migration, tube-formation ability and CRC cell proliferation. Additionally, in vivo studies showed that MSCs promoted tumor angiogenesis partially through IL-8. Taken together, these findings suggest that IL-8 secreted by MSCs promotes CRC angiogenesis and growth and can therefore serve as a potential novel therapeutic target.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                2 June 2017
                2017
                : 12
                : 6
                : e0178831
                Affiliations
                [1 ]Children’s Health Center, The Second Hospital, Nanjing Medical University, Nanjing, Jiangsu, P. R. China
                [2 ]Clinical Molecular Diagnostic Laboratory, The Second Hospital, Nanjing Medical University, Nanjing, Jiangsu, P. R. China
                [3 ]The Second Clinical School, Nanjing Medical University, Nanjing, Jiangsu, P. R.China
                [4 ]Center for Vascular and inflammatory Diseases, University of Maryland School of Medicine, Baltimore, MD, United States of America
                [5 ]State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, P. R.China
                Medical College of Wisconsin, UNITED STATES
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: WW ZH DZ.

                • Data curation: WW ZH.

                • Formal analysis: XJ JX XZ ZZ.

                • Funding acquisition: WW ZH DZ.

                • Investigation: WW ZH FW HG.

                • Methodology: WW ZH DZ.

                • Project administration: ZH DZ.

                • Resources: XZ ZZ.

                • Software: XJ JX HG.

                • Supervision: DZ.

                • Validation: ZH.

                • Visualization: WW ZH DZ.

                • Writing – original draft: WW ZH.

                • Writing – review & editing: ZH DZ.

                Article
                PONE-D-16-46729
                10.1371/journal.pone.0178831
                5456372
                28575053
                1fe9fc38-ff1a-42cc-a233-b0a6e797e3d1
                © 2017 Wu et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 24 November 2016
                : 19 May 2017
                Page count
                Figures: 5, Tables: 0, Pages: 15
                Funding
                Funded by: State Key Laboratory of Reproductive Medicine, Nanjing Medical University
                Award ID: SKLRM-KF-1202
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 81301822
                Award Recipient :
                Funded by: The Research Program funded by Nanjing Medical University
                Award ID: 2012NJMU088
                Award Recipient :
                Funded by: Key Project supported by Medical Science and technology development Foundation, Nanjing Department of Health
                Award ID: YKK13177
                Award Recipient :
                This work was supported by research grants from State Key Laboratory of Reproductive Medicine, Nanjing Medical University (SKLRM-KF-1202) to Weiling Wu, the National Natural Science Foundation of China (81301822) and the Research Program funded by Nanjing Medical University (2012NJMU088) to Zhenzhen Hu, Key Project supported by Medical Science and technology development Foundation, Nanjing Department of Health (YKK13177) to Datong Zheng. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
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