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Knockdown of miR-182 promotes apoptosis via regulating RIP1 deubiquitination in TNF-α-treated triple-negative breast cancer cells

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Tumor Biology

Abstract

Overexpression of microRNA-182 (miR-182) is found in multiple cancers, but the association of miR-182 expression with the sensitivity of triple-negative breast cancer (TNBC) cells to tumor necrosis factor-alpha (TNF-α) remains unknown. In this study, up-regulation of miR-182 was validated in TNBC patients and cell lines. Knockdown of miR-182 was observed to hinder the proliferation of BT-549 cells. More importantly, knockdown of miR-182 significantly promoted the apoptosis induced by TNF-α treatment in BT-549. JC-1 staining and western blot assays revealed that the K63-linked ubiquitin chains on receptor-interacting protein 1 (RIP1) were removed and the outer mitochondrial membrane potential (MMP) and permeability was altered upon combination of TNF-α with anti-miR-182. We then demonstrated that knockdown of miR-182 up-regulated the expression of cylindromatosis (CYLD) deubiquitinase, which promoted the formation of death-inducing signaling complex (DISC) and subsequent caspase-8 activation in TNF-α-treated BT-549 cells. Collectively, the results of the present study improve our understanding of the role of miR-182 in TNBC, knockdown of which facilitates the degradation of ubiquitin chains on RIP1, leading to the caspase-8 activation and apoptosis in TNF-α-treated TNBC cells. This may be valuable for the development of cancer therapy.

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Acknowledgments

This study is supported by the medical and health technology plan of Zhejiang Province (grant no.: 2015KYB268) and the TCM plan of Zhejiang Province (grant no.: 2010ZB038).

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Authors and Affiliations

  1. Department of Breast Surgery, First Affiliated Hospital of Zhejiang Chinese Medical University, Youdian Road 54#, Hangzhou, China

    Like Wo & Xidong Gu

  2. College of Life Science, Zhejiang Chinese Medical University, Hangzhou, 310053, China

    Dezhao Lu

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Correspondence to Xidong Gu.

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Wo, L., Lu, D. & Gu, X. Knockdown of miR-182 promotes apoptosis via regulating RIP1 deubiquitination in TNF-α-treated triple-negative breast cancer cells. Tumor Biol. 37, 13733–13742 (2016). https://doi.org/10.1007/s13277-016-5174-z

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