Overexpression of microRNA‑620 facilitates the resistance of triple negative breast cancer cells to gemcitabine treatment by targeting DCTD

Wu,Chao; Zhao,Aili; Tan,Tingzhao; Wang,Yuan; Shen,Zhentao

doi:10.3892/etm.2019.7601

Overexpression of microRNA‑620 facilitates the resistance of triple negative breast cancer cells to gemcitabine treatment by targeting DCTD

Authors:
- Chao Wu
- Aili Zhao
- Tingzhao Tan
- Yuan Wang
- Zhentao Shen
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Affiliations: Department of Medical Oncology, Liaocheng Cancer Prevention and Treatment Hospital, Liaocheng, Shandong 252000, P.R. China, Radiology Department, Liaocheng People's Hospital, Liaocheng, Shandong 252000, P.R. China
Published online on: May 23, 2019 https://doi.org/10.3892/etm.2019.7601
Pages: 550-558
Copyright: © Wu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Patients with triple negative breast cancer (TNBC) have a poor survival rate following chemotherapy due to drug resistance. Notably, the molecular mechanism of drug resistance remains elusive. Between December 2011 and December 2014, 36 TNBC samples were obtained from Liaocheng People's Hospital. Three gemcitabine‑resistant MDA‑MB‑231 cell lines (MDA‑MB‑231rGEM1, MDA‑MB‑231rGEM2 and MDA‑MB‑231rGEM3) were obtained by exposure of MDA‑MB‑231 cells to increasing concentrations of gemcitabine for >12 months. Reverse transcription‑quantitative polymerase chain reaction was performed to detect the expression levels of specific genes, including microRNA (miR)‑620, ATP‑binding cassette sub‑family B member 1 (ABCB1), ABCC10, cytidine monophosphate kinase, deoxycytidine monophosphate deaminase (DCTD), nucleoside diphosphate kinase 1 (NME1), ribonucleoside‑diphosphate reductase large subunit (RRM1) and RRMB2. Western blot analysis was performed to assess the protein expression levels of DCTD. Furthermore, cell proliferation was assessed using a Cell Counting Kit‑8 assay and cell apoptosis was detected using an Annexin V/Dead Cell Apoptosis kit. Interactions between miR‑620 and DCTD were predicted using TargetScan and detected with the dual luciferase reporter assay. Elevation of miR‑620 expression levels were detected in two of the assessed gemcitabine‑resistant MDA‑MB‑231 cell lines compared with MDA‑MB‑231 cells. Gemcitabine induced significant elevation of miR‑620 in MDA‑MB‑231 cells. An increase of DCTD at mRNA and protein expression levels in MDA‑MB‑231rGEM1 cells was observed compared with those in MDA‑MB‑231 cells. Results suggested that DCTD was directly regulated by miR‑620. Inhibition of miR‑620 and overexpression of DCTD reversed gemcitabine resistance in MDA‑MB‑231rGEM1 cells via inducing cell apoptosis and cell growth arrest. A negative correlation was identified between miR‑620 and DCTD mRNA expression levels in patients with TNBC. The present results demonstrated that overexpression of miR‑620 could contribute to the development of gemcitabine resistance in patients with TNBC via the direct downregulation of DCTD.

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1	DeSantis C, Ma J, Bryan L and Jemal A: Breast cancer statistics, 2013. CA Cancer J Clin. 64:52–62. 2014. View Article : Google Scholar : PubMed/NCBI
2	Dent R, Trudeau M, Pritchard KI, Hanna WM, Kahn HK, Sawka CA, Lickley LA, Rawlinson E, Sun P and Narod SA: Triple-negative breast cancer: Clinical features and patterns of recurrence. Clin Cancer Res. 13:4429–4434. 2007. View Article : Google Scholar : PubMed/NCBI
3	Wu ZH, Lin C, Liu MM, Zhang J, Tao ZH and Hu XC: Src inhibition can synergize with gemcitabine and reverse resistance in triple negative breast cancer cells via the AKT/c-Jun pathway. PLoS One. 11:e01692302016. View Article : Google Scholar : PubMed/NCBI
4	Chen M, He M, Song Y, Chen L, Xiao P, Wan X, Dai F and Shen P: The cytoprotective role of gemcitabine-induced autophagy associated with apoptosis inhibition in triple-negative MDA-MB-231 breast cancer cells. Int J Mol Med. 34:276–282. 2014. View Article : Google Scholar : PubMed/NCBI
5	de Sousa Cavalcante L and Monteiro G: Gemcitabine: Metabolism and molecular mechanisms of action, sensitivity and chemoresistance in pancreatic cancer. Eur J Pharmacol. 741:8–16. 2014. View Article : Google Scholar : PubMed/NCBI
6	Iorio MV and Croce CM: microRNA involvement in human cancer. Carcinogenesis. 33:1126–1133. 2012. View Article : Google Scholar : PubMed/NCBI
7	Hesse M and Arenz C: MicroRNA maturation and human disease. Methods Mol Biol. 1095:11–25. 2014. View Article : Google Scholar : PubMed/NCBI
8	Gommans WM and Berezikov E: Controlling miRNA regulation in disease. Methods Mol Biol. 822:1–18. 2012. View Article : Google Scholar : PubMed/NCBI
9	Selcuklu SD, Donoghue MT, Rehmet K, de Souza Gomes M, Fort A, Kovvuru P, Muniyappa MK, Kerin MJ, Enright AJ and Spillane C: MicroRNA-9 inhibition of cell proliferation and identification of novel miR-9 targets by transcriptome profiling in breast cancer cells. J Biol Chem. 287:29516–29528. 2012. View Article : Google Scholar : PubMed/NCBI
10	Zhu M, Xu Y, Ge M, Gui Z and Yan F: Regulation of UHRF1 by microRNA-9 modulates colorectal cancer cell proliferation and apoptosis. Cancer Sci. 106:833–839. 2015. View Article : Google Scholar : PubMed/NCBI
11	Pouliot LM, Chen YC, Bai J, Guha R, Martin SE, Gottesman MM and Hall MD: Cisplatin sensitivity mediated by WEE1 and CHK1 is mediated by miR-155 and the miR-15 family. Cancer Res. 72:5945–5955. 2012. View Article : Google Scholar : PubMed/NCBI
12	Lv J, Xia K, Xu P, Sun E, Ma J, Gao S, Zhou Q, Zhang M, Wang F, Chen F, et al: miRNA expression patterns in chemoresistant breast cancer tissues. Biomed Pharmacother. 68:935–942. 2014. View Article : Google Scholar : PubMed/NCBI
13	Park EY, Chang E, Lee EJ, Lee HW, Kang HG, Chun KH, Woo YM, Kong HK, Ko JY, Suzuki H, et al: Targeting of miR34a-NOTCH1 axis reduced breast cancer stemness and chemoresistance. Cancer Res. 74:7573–7582. 2014. View Article : Google Scholar : PubMed/NCBI
14	Chen L and Bourguignon LY: Hyaluronan-CD44 interaction promotes c-Jun signaling and miRNA21 expression leading to Bcl-2 expression and chemoresistance in breast cancer cells. Mol Cancer. 13:522014. View Article : Google Scholar : PubMed/NCBI
15	Chen X, Wang YW, Xing AY, Xiang S, Shi DB, Liu L, Li YX and Gao P: Suppression of SPIN1-mediated PI3K-Akt pathway by miR-489 increases chemosensitivity in breast cancer. J Pathol. 239:459–472. 2016. View Article : Google Scholar : PubMed/NCBI
16	Dorman SN, Baranova K, Knoll JH, Urquhart BL, Mariani G, Carcangiu ML and Rogan PK: Genomic signatures for paclitaxel and gemcitabine resistance in breast cancer derived by machine learning. Mol Oncol. 10:85–100. 2016. View Article : Google Scholar : PubMed/NCBI
17	Eriksson S, Skog S, Tribukait B and Jäderberg K: Deoxyribonucleoside triphosphate metabolism and the mammalian cell cycle. Effects of thymidine on wild-type and dCMP deaminase-deficient mouse S49 T-lymphoma cells. Exp Cell Res. 155:129–140. 1984. View Article : Google Scholar : PubMed/NCBI
18	Xu YZ and Plunkett W: Modulation of deoxycytidylate deaminase in intact human leukemia cells. Action of 2′,2′-difluorodeoxycytidine. Biochem Pharmacol. 44:1819–1827. 1992. View Article : Google Scholar : PubMed/NCBI
19	Tsaur I, Makarević J, Juengel E, Gasser M, Waaga-Gasser AM, Kurosch M, Reiter M, Wedel S, Bartsch G, Haferkamp A, et al: Resistance to the mTOR-inhibitor RAD001 elevates integrin α2- and β1-triggered motility, migration and invasion of prostate cancer cells. Br J Cancer. 107:847–855. 2012. View Article : Google Scholar : PubMed/NCBI
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
21	Lee SY, Im SA, Park YH, Woo SY, Kim S, Choi MK, Chang W, Ahn JS and Im YH: Genetic polymorphisms of SLC28A3, SLC29A1 and RRM1 predict clinical outcome in patients with metastatic breast cancer receiving gemcitabine plus paclitaxel chemotherapy. Eur J Cancer. 50:698–705. 2014. View Article : Google Scholar : PubMed/NCBI
22	O'Reilly EA, Gubbins L, Sharma S, Tully R, Guang MH, Weiner-Gorzel K, McCaffrey J, Harrison M, Furlong F, Kell M and McCann A: The fate of chemoresistance in triple negative breast cancer (TNBC). BBA Clin. 3:257–275. 2015. View Article : Google Scholar : PubMed/NCBI
23	Rosell R, Cobo M, Isla D, Camps C and Massuti B: Pharmacogenomics and gemcitabine. Ann Oncol. 17 (Suppl 5):v13–v16. 2006. View Article : Google Scholar : PubMed/NCBI
24	Rajabpour A, Afgar A, Mahmoodzadeh H, Radfar JE, Rajaei F and Teimoori-Toolabi L: MiR-608 regulating the expression of ribonucleotide reductase M1 and cytidine deaminase is repressed through induced gemcitabine chemoresistance in pancreatic cancer cells. Cancer Chemother Pharmacol. 80:765–775. 2017. View Article : Google Scholar : PubMed/NCBI
25	Zhuang J, Shen L, Yang L, Huang X, Lu Q, Cui Y, Zheng X, Zhao X, Zhang D, Huang R, et al: TGFβ1 promotes gemcitabine resistance through regulating the LncRNA-LET/NF90/miR-145 signaling axis in bladder cancer. Theranostics. 7:3053–3067. 2017. View Article : Google Scholar : PubMed/NCBI
26	Huang X, Taeb S, Jahangiri S, Korpela E, Cadonic I, Yu N, Krylov SN, Fokas E, Boutros PC and Liu SK: miR-620 promotes tumor radioresistance by targeting 15-hydroxyprostaglandin dehydrogenase (HPGD). Oncotarget. 6:22439–22451. 2015.PubMed/NCBI
27	Ueno H, Kiyosawa K and Kaniwa N: Pharmacogenomics of gemcitabine: Can genetic studies lead to tailor-made therapy? Br J Cancer. 97:145–151. 2007. View Article : Google Scholar : PubMed/NCBI
28	Hu H, Wang Z, Li M, Zeng F, Wang K, Huang R, Wang H, Yang F, Liang T, Huang H and Jiang T: Gene expression and methylation analyses suggest DCTD as a prognostic factor in malignant glioma. Sci Rep. 7:115682017. View Article : Google Scholar : PubMed/NCBI