Tumor Biology

, Volume 37, Issue 2, pp 1599–1607 | Cite as

MiR-944 functions as a novel oncogene and regulates the chemoresistance in breast cancer

Original Article

Abstract

MircroRNAs are emerging as critical regulators in carcinogenesis and chemoresistance in multiple cancer types. In this study, we observed that the miR-944 level was upregulated in breast cancer patients’ serum and tumor tissues, suggesting that miR-944 is a tumor promoter in breast cancer. To investigate the role of miR-944, we performed gain- and loss-of-function experiments in vitro. We then demonstrated that miR-944 promotes cell proliferation and tumor metastasis in breast cancer cell lines. Furthermore, we indicated that miR-944 is associated with cisplatin resistance by targeting BNIP3. Knockdown of the miR-944 by specific inhibitors significantly increased the cytotoxicity of cisplatin in cisplatin-resistant MCF-7 cells (MCF-7/R). Importantly, we found that the sensitization of miR-944 inhibitors to cisplatin cytotoxicity was abolished by BNIP3 siRNA which decreased the expression of BNIP3 gene. Finally, we demonstrated that miR-944 inhibitors promoted the loss of mitochondrial membrane potential (MMP) caused by cisplatin in MCF-7/R cells, resulting in the release of mitochondria-derived apoptogenic proteins into cytoplasm, and then, the caspase-3 was activated. In summary, our study showed that miR-944 functions as a novel oncogene and regulates the cisplatin resistance in breast cancer. The miR-944-BNIP3-MMP-caspase-3 pathway might be a novel target for the chemotherapy of breast cancer.

Keywords

Breast cancer MiR-944 Oncogene BNIP3 Cisplatin resistance 

Notes

Acknowledgments

Thanks are due to Medical and health research projects in Zhejiang Province (grant no: 2014KYA094) and The Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education for supporting this study

Compliance with ethical standards

The study was approved by Ethic Committee of The Second Affiliated Hospital, School of medicine, Zhejiang University.

Author contributions

KJ designed the study. KJ and HH wrote the manuscript. HH and WT performed out the immunohistochemistry and the related statistical analysis. HH, HC, and KJ carried out the cell culture and transfection, Western blot, RT-PCR, and flow Ccytometry. All authors approved the final version of the manuscript.

Conflicts of interest

The authors have no conflicts of interest.

References

  1. 1.
    Siegel R, Naishadham D, Jemal A. Cancer statistics, 2013. CA Cancer J Clin. 2013;63:11–30.CrossRefPubMedGoogle Scholar
  2. 2.
    Jerusalem G, Rorive A, Collignon J. Chemotherapy options for patients suffering from heavily pretreated metastatic breast cancer. Future Oncol. 2015;11:1775–89.CrossRefPubMedGoogle Scholar
  3. 3.
    Yao YS, Qiu WS, Yao RY, Zhang Q, Zhuang LK, Zhou F, et al. miR-141 confers docetaxel chemoresistance of breast cancer cells via regulation of EIF4E expression. Oncol Rep. 2015;33:2504–12.PubMedGoogle Scholar
  4. 4.
    Arslan C, Ozdemir E, Dogan E, Ozisik Y, Altundag K. Secondary hematological malignancies after treatment of non-metastatic breast cancer. J BUON. 2011;16:744–50.PubMedGoogle Scholar
  5. 5.
    Mohell N, Alfredsson J, Fransson Å, Uustalu M, Byström S, Gullbo J, et al. APR-246 overcomes resistance to cisplatin and doxorubicin in ovarian cancer cells. Cell Death Dis. 2015;6, e1794.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Gao JH, Chen FH, Wang L, Wei H, Meng SL. YM155 inhibits tumor growth and enhances chemosensitivity to cisplatin in osteosarcoma. Eur Rev Med Pharmacol Sci. 2015;19:2062–9.PubMedGoogle Scholar
  7. 7.
    Ambros V. MicroRNA pathways in flies and worms: growth, death, fat, stress, and timing. Cell. 2003;113:673–6.CrossRefPubMedGoogle Scholar
  8. 8.
    Nair N, Gongora E. MicroRNAs as therapeutic targets in cardiomyopathies: myth or reality? Biomol Concepts. 2014;5:439–48.CrossRefPubMedGoogle Scholar
  9. 9.
    Wolfson B, Eades G, Zhou Q. Roles of microRNA-140 in stem cell-associated early stage breast cancer. World J Stem Cells. 2014;6:591–7.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Mo ZH, Wu XD, Li S, Fei BY, Zhang B. Expression and Clinical Significance of MicroRNA-376a in Colorectal Cancer. Asian Pac J Cancer Prev. 2014;15:9523–7.CrossRefPubMedGoogle Scholar
  11. 11.
    Toraih EA, Mohammed EA, Farrag S, Ramsis N, Hosny S. Pilot Study of Serum MicroRNA-21 as a Diagnostic and Prognostic Biomarker in Egyptian Breast Cancer Patients. Mol Diagn Ther. 2015;19:179–90.CrossRefPubMedGoogle Scholar
  12. 12.
    Kaboli PJ, Rahmat A, Ismail P, Ling KH. MicroRNA-based therapy and breast cancer: A comprehensive review of novel therapeutic strategies from diagnosis to treatment. Pharmacol Res. 2015;97:104–21.CrossRefPubMedGoogle Scholar
  13. 13.
    Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 2001;25:402–8.CrossRefPubMedGoogle Scholar
  14. 14.
    Chen C, Ridzon DA, Lee DH, et al. Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res. 2005;33, e179.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Wang J, Tian X, Han R, Zhang X, Wang X, Shen H, et al. Downregulation of miR-486-5p contributes to tumor progression and metastasis by targeting protumorigenic ARHGAP5 in lung cancer. Oncogene. 2014;33:1181–9.CrossRefPubMedGoogle Scholar
  16. 16.
    Gu JW, Bailey AP, Sartin A, Makey I, Brady AL. Ethanol stimulates tumor progression and expression of vascular endothelial growth factor in chick embryos. Cancer. 2005;103:422–31.CrossRefPubMedGoogle Scholar
  17. 17.
    Chen H, Zhu G, Li Y, Padia RN, Dong Z, Pan ZK, et al. Extracellular signal–regulated kinase signaling pathway regulates breast cancer cell migration by maintaining slug expression. Cancer Res. 2009;69:9228–35.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Prathapan A, Vineetha VP, Raghu KG. Protective effect of Boerhaavia diffusa L. against mitochondrial dysfunction in angiotensin II induced hypertrophy in H9c2 cardiomyoblast cells. PLoS One. 2014;9, e96220.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Ray R, Chen G, Vande Velde C, et al. BNIP3 heterodimerizes with Bcl-2/Bcl-X(L) and induces cell death independent of a Bcl-2 homology 3(BH3) domain at both mitochondrial and nonmitochondrial sites. J Biol Chem. 2000;275:1439–48.CrossRefPubMedGoogle Scholar
  20. 20.
    Kaza N, Kohli L, Roth KA, et al. BNIP3 regulates AT101 [(−)-gossypol] induced death in malignant peripheral nerve sheath tumor cells. PLoS One. 2014;9, e96733.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Witten D, Tibshirani R, Gu SG, et al. Ultra-high throughput sequencing-based small RNA discov ery and discrete statistical biomarker analysis in a collection of cervical tumours and matched controls. BMC Biol. 2010;8:58.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Christensen LL, Tobiasen H, Holm A, et al. MiRNA-362-3p induces cell cycle arrest through targeting of E2F1, USF2 and PTPN1 and is associated with recurrence of colorectal cancer. Int J Cancer. 2013;133:67–78.CrossRefPubMedGoogle Scholar
  23. 23.
    Nordentoft I, Birkenkamp-Demtroder K, Agerbaek M, et al. miRNAs associated with chemo-sensitivity in cell lines and in advanced bladder cancer. BMC Med Genomics. 2012;5:40.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Yin W, Nie Y, Zhang Z, Xie L, He X. miR-193b acts as a cisplatin sensitizer via the caspase-3-dependent pathway in HCC chemotherapy. Oncol Rep. 2015;34:368–74.PubMedGoogle Scholar
  25. 25.
    Weiner-Gorzel K, Dempsey E, Milewska M, McGoldrick A, Toh V, Walsh A, et al. Overexpression of the microRNA miR-433 promotes resistance to paclitaxel through the induction of cellular senescence in ovarian cancer cells. Cancer Med. 2015;4:745–58.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Jiang C, Long J, Liu B, Xie X, Kuang M. Mcl-1 Is a Novel Target of miR-26b That Is Associated with the Apoptosis Induced by TRAIL in HCC Cells. Biomed Res Int. 2015;2015:572738.PubMedPubMedCentralGoogle Scholar
  27. 27.
    Ray R, Chen G, Vande Velde C, et al. BNIP3 heterodimerizes with Bcl-2/Bcl-X(L) and induces cell death independent of a Bcl-2 homology 3(BH3) domain at both mitochondrial and nonmitochondrial sites. J Biol Chem. 2000;275:1439–48.CrossRefPubMedGoogle Scholar
  28. 28.
    Gustafsson AB. Bnip3 as a dual regulator of mitochondrial turnover and cell death in the myocardium. Pediatr Cardiol. 2011;32:267–74.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Rikka S, Quinsay MN, Thomas RL, Kubli DA, Zhang X, Murphy AN, et al. Bnip3 impairs mitochondrial bioenergetics and stimulates mitochondrial turnover. Cell Death Differ. 2011;18:721–31.CrossRefPubMedGoogle Scholar
  30. 30.
    Erkan M, Kleeff J, Esposito I, Giese T, Ketterer K, Büchler MW, et al. Loss of BNIP3 expression is a late event in pancreatic cancer contributing to chemoresistance and worsened prognosis. Oncogene. 2005;24:4421–32.CrossRefPubMedGoogle Scholar
  31. 31.
    Zu Y, Yang Z, Tang S, Han Y, Ma J. Effects of p-glycoprotein and its inhibitors on apoptosis in k562 cells. Molecules. 2014;19:13061–75.CrossRefPubMedGoogle Scholar
  32. 32.
    Zhao JX, Liu H, Lv J, Yang XJ. Wortmannin enhances cisplatin-induced apoptosis in human ovarian cancer cells in vitro. Eur Rev Med Pharmacol Sci. 2014;18:2428–34.PubMedGoogle Scholar
  33. 33.
    Tian HY, Li ZX, Li HY, Wang HJ, Zhu XW, Dou ZH. Effects of 14 single herbs on the induction of caspase-3 in tumor cells: a brief review. Chin J Integr Med. 2013;19:636–40.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Haifei He
    • 1
  • Wei Tian
    • 1
    • 2
  • Hailong Chen
    • 1
    • 2
  • Kai Jiang
    • 3
  1. 1.Department of Surgical OncologyThe Second Affiliated Hospital, School of medicine, Zhejiang UniversityHangzhouChina
  2. 2.Cancer Institute (The Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education)The Second Affiliated Hospital, School of medicine, Zhejiang UniversityHangzhouChina
  3. 3.Department of Clinical LaboratoryTongde Hospital of Zhejiang ProvinceHangzhou city provinceChina

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