Summary
miR-200c has been shown to regulate the epithelial-mesenchymal transition (EMT) by inhibiting ZEB1 and ZEB2 expression in breast cancer cells. This study further examined the role of miR-200c in the invasion and metastasis of breast cancer that goes beyond the regulation on ZEB1 and ZEB2 expression. In this study, the bioinformatics software (miRanda) was used to predict the target gene of miR-200c and Renilla luciferase assay to verify the result. The metastatic breast cancer cells MDA-MB-231 were cultured and transfected with the miR-200c mimic or inhibitor. The expressions of miR-200c and HMGB1 were detected by RT-PCR and Western blotting, respectively. Transwell assay and wound healing assay were employed to examine the invasive and migrating ability of transfected cells. Target prediction and Renilla luciferase analysis revealed that HMGB1 was a putative target gene of miR-200c. After transfection of MDA-MB-231 cells with the miR-200c mimic or inhibitor, the expression of miR-200c was significantly increased or decreased when compared with cells transfected with the miR-200c mimic NC or inhibitor NC. Moreover, the expression of HMGB1 was reversely correlated with that of miR-200c in transfected cells. Tranwell assay showed that the number of invasive cells was significantly reduced in miR-200c mimic group when compared with miR-200c inhibitor group. It was also found that the migrating ability of cells transfected with miR-200c mimics was much lower than that of cells transfected with miR-200c inhibitors. It was suggested that miR-200c can suppress the invasion and migration of breast cancer cells by regulating the expression of HMGB1. miR-200c and HMGB1 may become useful biomarkers for progression of breast cancer and targets of gene therapy.
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References
Siegel R, Naishadham D, Jemal A. Cancer statistics, 2013. CA Cancer J Clin, 2013,63(1):11–30
Jiang P, Enomoto A, Takahashi M. Cell biology of the movement of breast cancer cells: intracellular signaling and the actin cytoskeleton. Cancer Lett, 2009,284(2):122–130
Graves P, Zeng Y. Biogenesis of mammalian microRNAs: a global view. Genomics Proteomics Bioinformatics, 2012,10(5):239–245
Macfarlane LA, Murphy PR. MicroRNA: biogenesis, function and role in cancer. Curr Genomics, 2010,11(7):537–561
Burk U, Schubert J, Wellner U, et al. A reciprocal repression between ZEB1 and members of the miR-200 family promotes EMT and invasion in cancer cells. EMBO Rep, 2008,9(6):582–589
Peter ME. Let-7 and miR-200 microRNAs: guardians against pluripotency and cancer progression. Cell Cycle, 2009,8(6):843–852
Shimono Y, Zabala M, Cho RW, et al. Downregulation of miRNA-200c links breast cancer stem cells with normal stem cells. Cell, 2009,138(3):592–603
Schickel R, Park SM, Murmann AE, et al. mir-200c regulates induction of apoptosis through CD95 by targeting FAP-1. Mol Cell, 2010,38(6):908–915
Chang CJ, Chao CH, Xia W, et al. p53 regulates epithelial-mesenchymal transition and stem cell properties through modulating miRNAs. Nat Cell Biol, 2011,13(3):317–323
Cochrane DR, Spoelstra NS, Howe EN, et al. MicroRNA-200c mitigates invasiveness and restores sensitivity to microtubule-targeting chemotherapeutic agents. Mol Cancer Ther, 2009,8(5):1055–1066
Lin J, Liu C, Gao F, et al. miR-200c enhances radiosensitivity of human breast cancer cells. J Cell Biochem, 2013,114(3):606–615
Ahmad A, Aboukameel A, Kong D, et al. Phosphoglucose isomerase/autocrine motility factor mediates epithelial-mesenchymal transition regulated by miR-200 in breast cancer cells. Cancer Res, 2011,71(9):3400–3409
Howe EN, Cochrane DR, Cittelly DM, et al. miR-200c targets a NF-κB up-regulated TrkB/NTF3 autocrine signaling loop to enhance anoikis sensitivity in triple negative breast cancer. PLoS One, 2012,7(11):e49987
Jurmeister S, Baumann M, Balwierz A, et al. MicroRNA-200c represses migration and invasion of breast cancer cells by targeting actin-regulatory proteins FHOD1 and PPM1F. Mol Cell Biol, 2012,32(3):633–651
Chen Y, Sun Y, Chen L, et al. miRNA-200c increases the sensitivity of breast cancer cells to doxorubicin through the suppression of E-cadherin-mediated PTEN/Akt signaling. Mol Med Rep, 2013,7(5):1579–1584
Nikoletopoulou V, Markaki M, Palikaras K, et al. Crosstalk between apoptosis, necrosis and autophagy. Biochim Biophys Acta, 2013,1833(12):3448–3459
Ko YB, Kim BR, Nam SL, et al. High-mobility group box 1 (HMGB1) protein regulates tumor-associated cell migration through the interaction with BTB domain. Cell Signal, 2014,26(4):777–783
Tang D, Kang R, Zeh HJ 3rd, et al. High-mobility group box 1 and cancer. BBA, 2010,1799(1–2):131–140
Sparatore B, Patrone M, Passalacqua M, et al. Activation of A431 human carcinoma cell motility by extracellular high-mobility group box 1 protein and epidermal growth factor stimuli. Biochem J, 2005,389(1):215–221
Brezniceanu ML, Völp K, Bösser S, et al. HMGB1 inhibits cell death in yeast and mammalian cells and is abundantly expressed in human breast carcinoma. FASEB J, 2003,17(10):1295–1297
Livesey KM, Kang R, Vernon P, et al. p53/HMGB1 complexes regulate autophagy and apoptosis. Cancer Res, 2012,72(8):1996–2005
van Beijnum JR, Nowak-Sliwinska P, van den Boezem E, et al. Tumor angiogenesis is enforced by autocrine regulation of high-mobility group box 1. Oncogene, 2013,32(3):363–374
Jiao Y, Wang HC, Fan SJ. Growth suppression and radiosensitivity increase by HMGBI in breast cancer. Acta pharmacologica Sinica, 2007,28(12):1957–1967
Bernardini M, Lee CH, Beheshti B, et a1. High-resolution mapping of genomic imbalance and identification of gene expression profiles associated with differential chemotherapy response in serous epithelial ovarian cancer. Neoplasia, 2005,7(6):603–613
Barnes KR, Kutikov A, Lippard SJ. Synthesis, characterization, and cytotoxicity of a series of estrogen-tethered platinum (IV) complexes. Chem Biol, 2004,11(4):557–564
Gregory PA, Bracken CP, Smith E, et al. An autocrine TGF-beta/ZEB/miR-200 signaling network regulates establishment and maintenance of epithelial-mesenchymal transition. Mol Biol Cell, 2011,22(10):1686–1698
Tobar N, Villar V, Santibanez JF. ROS-NFkappaB mediates TGF-beta1-induced expression of urokinase-type plasminogen activator, matrix metalloproteinase-9 and cell invasion. Mol Cell Biochem, 2010,340(1–2):195–202
Chua HL, Bhat-Nakshatri P, Clare SE. NF-kappaB represses E-cadherin expression and enhances epithelial to mesenchymal transition of mammary epithelial cells: potential involvement of ZEB-1 and ZEB-2. Oncogene, 2007,26(5):711–724
Wu X, Mi Y, Yang H, et al. The activation of HMGB 1 as a progression factor on inflammation response in normal human bronchial epithelial cells through RAGE/JNK/NF-κB pathway, Mol Cell Biochem, 2013,380(1–2):249–257
Smolarczyk R, Cichoń T, Jarosz M, et al. HMGB1—its role in tumor progression and anticancer therapy. Postepy Hig Med Dosw, 2012,22(66):913–920
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The project was supported by the Innovation Foundation of Excellent Intellectuals in Henan Province (No. 2109901).
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Chang, Bp., Wang, Ds., Xing, Jw. et al. miR-200c inhibits metastasis of breast cancer cells by targeting HMGB1. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 34, 201–206 (2014). https://doi.org/10.1007/s11596-014-1259-3
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DOI: https://doi.org/10.1007/s11596-014-1259-3