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Expression and regulatory function of miRNA-182 in triple-negative breast cancer cells through its targeting of profilin 1

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Tumor Biology

Abstract

We aimed to evaluate the expression of microRNA-182 (miR-182) in triple-negative breast cancer (TNBC) tissues and the TNBC cell line MDA-MB-231 and to investigate the effects of mirR-182 on the cellular behavior of MDA-MB-231 and the expression of the target gene profilin 1 (PFN1), thus providing new methods and new strategies for the treatment of TNBC. Quantitative real-time PCR (qRT-PCR) was utilized to evaluate the expression of miR-182 in TNBC tissues, relatively normal tissues adjacent to TNBC and the TNBC cell line MDA-MB-231. Forty-eight hours after the MDA-MB-231 cells were transfected with the miR-182 inhibitor, qRT-PCR was utilized to detect the changes in miR-182 expression levels, and an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was utilized to determine the effects of miR-182 on cell viability. Flow cytometry was adopted to determine whether miR-182 affects the proliferation rates and apoptosis levels of the MDA-MB-231 cells. The transwell migration assay method was used to investigate the effects of miR-182 on the migration of the MDA-MB-231 cells. A luciferase reporter gene system was applied to validate that PFN1 was the target gene of miR-182. Western blot was used to measure the effects of miR-182 on the PFN1 protein expression levels in the cells. qRT-PCR results showed that compared with the relatively normal tissues adjacent to TNBC, miR-182 expression was significantly increased in the TNBC tissues and the MDA-MB-231 cells (p < 0.01). Compared with the control group, MDA-MB-231 cells transfected with the miR-182 inhibitor and incubated for 48 h showed significantly decreased miR-182 expression (p < 0.01). The results of an MTT assay showed that inhibition of miR-182 in MDA-MB-231 cells led to significantly reduced cell viability (p < 0.05). Flow cytometry analysis indicated that inhibition of miR-182 expression resulted in significantly decreased cell proliferation (p < 0.05) and significantly increased levels of apoptosis (p < 0.05). The results of a transwell migration assay showed that after inhibited of miR-182 expression, the number of cells passing through the transwell membranes was significantly decreased (p < 0.05). The results from a luciferase reporter gene system showed that compared with the control group, the relative luciferase activity of the group transfected with the miR-182 inhibitor was significantly increased (p < 0.05). Western blot analysis showed that compared with the control group, PFN1 protein expression levels were significantly increased in the MDA-MB-231 cells transfected with the miR-182 inhibitor and incubated for 48 h (p < 0.05). In conclusion, miR-182 is upregulated in TNBC tissues and cells. It promotes the proliferation and invasion of MDA-MB-231 cells and could negatively regulate PFN1 protein expression. Treatment strategies utilizing inhibition of miR-182 expression or overexpression of the PFN1 gene might benefit patients with TNBC.

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Acknowledgement

This study was supported by National Natural Science Foundation of China (No. 81150011).

Conflicts of interest

The authors have no commercial, proprietary, or financial interest in the products or companies described in this article.

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Authors and Affiliations

  1. Division of Breast Surgery, Department of General Surgery, General Hospital of Chinese People’s Liberation Army (301 Hospital), No. 28, Fuxing Rd, Beijing, 100853, People’s Republic of China

    Hailing Liu, Yan Wang, Yan-jun Zhang, Jie Li, Yi-qiong Zheng & Xi-ru Li

  2. Department of Endocrinology, General Hospital of Chinese People’s Liberation Army (301 Hospital), Beijing, China

    Xin Li

  3. Department of Pathology, General Hospital of Chinese People’s Liberation Army (301 Hospital), Beijing, China

    Mei Liu & Xin Song

Authors
  1. Hailing Liu
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  2. Yan Wang
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  3. Xin Li
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  4. Yan-jun Zhang
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  5. Jie Li
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  6. Yi-qiong Zheng
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Correspondence to Xi-ru Li.

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Hailing Liu, Yan Wang, and Xin Li contributed equally to this article.

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Liu, H., Wang, Y., Li, X. et al. Expression and regulatory function of miRNA-182 in triple-negative breast cancer cells through its targeting of profilin 1. Tumor Biol. 34, 1713–1722 (2013). https://doi.org/10.1007/s13277-013-0708-0

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  • DOI: https://doi.org/10.1007/s13277-013-0708-0

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