miR-106b~25 cluster regulates multidrug resistance in an ABC transporter-independent manner via downregulation of EP300

Hu,Yunhui; Li,Kaiyong; Asaduzzaman,Muhammad; Cuella,Raquel; Shi,Hui; Raguz,Selina; Coombes,Raoul  Charles; Zhou,Yuan; Yagüe,Ernesto

doi:10.3892/or.2015.4412

miR-106b~25 cluster regulates multidrug resistance in an ABC transporter-independent manner via downregulation of EP300

Authors:
- Yunhui Hu
- Kaiyong Li
- Muhammad Asaduzzaman
- Raquel Cuella
- Hui Shi
- Selina Raguz
- Raoul Charles Coombes
- Yuan Zhou
- Ernesto Yagüe
View Affiliations

Affiliations: The Third Department of Breast Cancer, China Tianjin Breast Cancer Prevention, Treatment and Research Center, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Tianjin 300060, P.R. China, Division of Cancer, Imperial College London, Hammersmith Hospital Campus, London W12 0NN, UK, State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, P.R. China, Division of Clinical Sciences, Imperial College London, Hammersmith Hospital Campus, London W12 0NN, UK
Published online on: November 12, 2015 https://doi.org/10.3892/or.2015.4412
Pages: 1170-1178

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

MicroRNA (miR)-106b~25 cluster regulates bypass of doxorubicin and γ-radiation induced senescence by downregulation of the E-cadherin transcriptional activator EP300. We asked whether upregulation of miR-106~25 cluster generates cells with a truly multidrug resistant (MDR) phenotype and whether this is due to upregulation of the ATP-binding cassette (ABC) transporter P-glycoprotein. We used minimally transformed mammary epithelial breast cancer cells (MTMECs) in which the miR-106b~25 cluster was experimentally upregulated by lentiviral transfection or in which hairpins targeting either EP300 or E-cadherin mRNAs have been expressed with lentiviruses. We find that overexpression of miR-106b~25 cluster led to the generation of MDR MTMECs (resistant to etoposide, colchicine and paclitaxel). Paclitaxel resistance was also studied after experimental downregulation of EP300 or E-cadherin. However none of these cells overexpressed P-glycoprotein or where able to efflux a fluorescent derivative of paclitaxel, making this phenotype drug-transporter independent. Paclitaxel treatment in MTMECs led to an increase in early apoptotic cells (Annexin V-positive), activation of caspase-9 and increase in the proportion of cells at the G2/M phase of the cell cycle. However, MTMEC overexpressing miR-106b~25 cluster, or with EP300 or E-cadherin downregulated, showed less activation of apoptosis, caspase-9 and caspase-3/-7 activities. Thus, miR-106b~25 cluster controls transporter-independent MDR by apoptosis evasion via downregulation of EP300.

View Figures

View References

1	Raguz S and Yagüe E: Resistance to chemotherapy: New treatments and novel insights into an old problem. Br J Cancer. 99:387–391. 2008. View Article : Google Scholar : PubMed/NCBI
2	Gottesman MM: Mechanisms of cancer drug resistance. Annu Rev Med. 53:615–627. 2002. View Article : Google Scholar : PubMed/NCBI
3	Gillet JP and Gottesman MM: Overcoming multidrug resistance in cancer: 35 years after the discovery of ABCB1. Drug Resist Updat. 15:2–4. 2012. View Article : Google Scholar : PubMed/NCBI
4	Gottesman MM, Fojo T and Bates SE: Multidrug resistance in cancer: Role of ATP-dependent transporters. Nat Rev Cancer. 2:48–58. 2002. View Article : Google Scholar : PubMed/NCBI
5	Steinbach D and Legrand O: ABC transporters and drug resistance in leukemia: Was P-gp nothing but the first head of the Hydra? Leukemia. 21:1172–1176. 2007. View Article : Google Scholar : PubMed/NCBI
6	Trock BJ, Leonessa F and Clarke R: Multidrug resistance in breast cancer: A meta-analysis of MDR1/gp170 expression and its possible functional significance. J Natl Cancer Inst. 89:917–931. 1997. View Article : Google Scholar : PubMed/NCBI
7	Clarke R, Leonessa F and Trock B: Multidrug resistance/P-glycoprotein and breast cancer: Review and meta-analysis. Semin Oncol. 32(Suppl 7): S9–S15. 2005. View Article : Google Scholar : PubMed/NCBI
8	Reinhart BJ, Slack FJ, Basson M, Pasquinelli AE, Bettinger JC, Rougvie AE, Horvitz HR and Ruvkun G: The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans. Nature. 403:901–906. 2000. View Article : Google Scholar : PubMed/NCBI
9	Negrini M and Calin GA: Breast cancer metastasis: A microRNA story. Breast Cancer Res. 10:2032008. View Article : Google Scholar : PubMed/NCBI
10	Jang JS, Jeon HS, Sun Z, Aubry MC, Tang H, Park CH, Rakhshan F, Schultz DA, Kolbert CP, Lupu R, et al: Increased miR-708 expression in NSCLC and its association with poor survival in lung adenocarcinoma from never smokers. Clin Cancer Res. 18:3658–3667. 2012. View Article : Google Scholar : PubMed/NCBI
11	Yang H, Kong W, He L, Zhao JJ, O'Donnell JD, Wang J, Wenham RM, Coppola D, Kruk PA, Nicosia SV, et al: MicroRNA expression profiling in human ovarian cancer: miR-214 induces cell survival and cisplatin resistance by targeting PTEN. Cancer Res. 68:425–433. 2008. View Article : Google Scholar : PubMed/NCBI
12	Zhou M, Liu Z, Zhao Y, Ding Y, Liu H, Xi Y, Xiong W, Li G, Lu J, Fodstad O, et al: MicroRNA-125b confers the resistance of breast cancer cells to paclitaxel through suppression of pro-apoptotic Bcl-2 antagonist killer 1 (Bak1) expression. J Biol Chem. 285:21496–21507. 2010. View Article : Google Scholar : PubMed/NCBI
13	Miller TE, Ghoshal K, Ramaswamy B, Roy S, Datta J, Shapiro CL, Jacob S and Majumder S: MicroRNA-221/222 confers tamoxifen resistance in breast cancer by targeting p27Kip1. J Biol Chem. 283:29897–29903. 2008. View Article : Google Scholar : PubMed/NCBI
14	Poliseno L, Salmena L, Riccardi L, Fornari A, Song MS, Hobbs RM, Sportoletti P, Varmeh S, Egia A and Fedele G: Identification of the miR-106b~25 microRNA cluster as a proto-oncogenic PTEN-targeting intron that cooperates with its host gene MCM7 in transformation. Sci Signal. 3:ra292010. View Article : Google Scholar : PubMed/NCBI
15	Petrocca F, Visone R, Onelli MR, Shah MH, Nicoloso MS, de Martino I, Iliopoulos D, Pilozzi E, Liu CG, Negrini M, et al: E2F1-regulated microRNAs impair TGFbeta-dependent cell-cycle arrest and apoptosis in gastric cancer. Cancer Cell. 13:272–286. 2008. View Article : Google Scholar : PubMed/NCBI
16	Acosta JC and Gil J: Senescence: A new weapon for cancer therapy. Trends Cell Biol. 22:211–219. 2012. View Article : Google Scholar : PubMed/NCBI
17	Zhou Y, Hu Y, Yang M, Jat P, Li K, Lombardo Y, Xiong D, Coombes RC, Raguz S and Yagüe E: The miR-106b~25 cluster promotes bypass of doxorubicin-induced senescence and increase in motility and invasion by targeting the E-cadherin transcriptional activator EP300. Cell Death Differ. 21:462–474. 2014. View Article : Google Scholar :
18	Zhao JJ, Gjoerup OV, Subramanian RR, Cheng Y, Chen W, Roberts TM and Hahn WC: Human mammary epithelial cell transformation through the activation of phosphatidylinositol 3-kinase. Cancer Cell. 3:483–495. 2003. View Article : Google Scholar : PubMed/NCBI
19	Unsworth H, Raguz S, Edwards HJ, Higgins CF and Yagüe E: mRNA escape from stress granule sequestration is dictated by localization to the endoplasmic reticulum. FASEB J. 24:3370–3380. 2010. View Article : Google Scholar : PubMed/NCBI
20	Lee JS, Paull K, Alvarez M, Hose C, Monks A, Grever M, Fojo AT and Bates SE: Rhodamine efflux patterns predict P-glycoprotein substrates in the National Cancer Institute drug screen. Mol Pharmacol. 46:627–638. 1994.PubMed/NCBI
21	Yagüe E, Arance A, Kubitza L, O'Hare M, Jat P, Ogilvie CM, Hart IR, Higgins CF and Raguz S: Ability to acquire drug resistance arises early during the tumorigenesis process. Cancer Res. 67:1130–1137. 2007. View Article : Google Scholar : PubMed/NCBI
22	Yague E, Armesilla AL, Harrison G, Elliott J, Sardini A, Higgins CF and Raguz S: P-glycoprotein (MDR1) expression in leukemic cells is regulated at two distinct steps, mRNA stabilization and translational initiation. J Biol Chem. 278:10344–10352. 2003. View Article : Google Scholar : PubMed/NCBI
23	Raguz S, Adams C, Masrour N, Rasul S, Papoutsoglou P, Hu Y, Cazzanelli G, Zhou Y, Patel N, Coombes C, et al: Loss of O⁶-methylguanine-DNA methyltransferase confers collateral sensitivity to carmustine in topoisomerase II-mediated doxo-rubicin resistant triple negative breast cancer cells. Biochem Pharmacol. 85:186–196. 2013. View Article : Google Scholar
24	Vichai V and Kirtikara K: Sulforhodamine B colorimetric assay for cytotoxicity screening. Nat Protoc. 1:1112–1116. 2006. View Article : Google Scholar
25	Hu Y, Cheng X, Li S, Zhou Y, Wang J, Cheng T, Yang M and Xiong D: Inhibition of sorcin reverses multidrug resistance of K562/A02 cells and MCF-7/A02 cells via regulating apoptosis-related proteins. Cancer Chemother Pharmacol. 72:789–798. 2013. View Article : Google Scholar : PubMed/NCBI
26	Rasul S, Balasubramanian R, Filipović A, Slade MJ, Yagüe E and Coombes RC: Inhibition of gamma-secretase induces G2/M arrest and triggers apoptosis in breast cancer cells. Br J Cancer. 100:1879–1888. 2009. View Article : Google Scholar : PubMed/NCBI
27	Nitiss JL: Targeting DNA topoisomerase II in cancer chemotherapy. Nat Rev Cancer. 9:338–350. 2009. View Article : Google Scholar : PubMed/NCBI
28	Ewald JA, Desotelle JA, Wilding G and Jarrard DF: Therapy-induced senescence in cancer. J Natl Cancer Inst. 102:1536–1546. 2010. View Article : Google Scholar : PubMed/NCBI
29	McGrogan BT, Gilmartin B, Carney DN and McCann A: Taxanes, microtubules and chemoresistant breast cancer. Biochim Biophys Acta. 1785:96–132. 2008.
30	Wu CP, Hsieh CH and Wu YS: The emergence of drug transporter-mediated multidrug resistance to cancer chemotherapy. Mol Pharm. 8:1996–2011. 2011. View Article : Google Scholar : PubMed/NCBI
31	Friedrich K, Wieder T, Von Haefen C, Radetzki S, Jänicke R, Schulze-Osthoff K, Dörken B and Daniel PT: Overexpression of caspase-3 restores sensitivity for drug-induced apoptosis in breast cancer cell lines with acquired drug resistance. Oncogene. 20:2749–2760. 2001. View Article : Google Scholar : PubMed/NCBI
32	Holohan C, Van Schaeybroeck S, Longley DB and Johnston PG: Cancer drug resistance: An evolving paradigm. Nat Rev Cancer. 13:714–726. 2013. View Article : Google Scholar : PubMed/NCBI
33	Kathawala RJ, Gupta P, Ashby CR Jr and Chen ZS: The modulation of ABC transporter-mediated multidrug resistance in cancer: A review of the past decade. Drug Resist Updat. 18:1–17. 2015. View Article : Google Scholar : PubMed/NCBI
34	May CD, Sphyris N, Evans KW, Werden SJ, Guo W and Mani SA: Epithelial-mesenchymal transition and cancer stem cells: A dangerously dynamic duo in breast cancer progression. Breast Cancer Res. 13:2022011. View Article : Google Scholar : PubMed/NCBI
35	Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY, Brooks M, Reinhard F, Zhang CC, Shipitsin M, et al: The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell. 133:704–715. 2008. View Article : Google Scholar : PubMed/NCBI
36	Korpal M, Lee ES, Hu G and Kang Y: The miR-200 family inhibits epithelial-mesenchymal transition and cancer cell migration by direct targeting of E-cadherin transcriptional repressors ZEB1 and ZEB2. J Biol Chem. 283:14910–14914. 2008. View Article : Google Scholar : PubMed/NCBI
37	Ferreira P, Oliveira MJ, Beraldi E, Mateus AR, Nakajima T, Gleave M, Yokota J, Carneiro F, Huntsman D, Seruca R, et al: Loss of functional E-cadherin renders cells more resistant to the apoptotic agent taxol in vitro. Exp Cell Res. 310:99–104. 2005. View Article : Google Scholar : PubMed/NCBI
38	Shiota M, Yokomizo A, Kashiwagi E, Tada Y, Inokuchi J, Tatsugami K, Kuroiwa K, Uchiumi T, Seki N and Naito S: Foxo3a expression and acetylation regulate cancer cell growth and sensitivity to cisplatin. Cancer Sci. 101:1177–1185. 2010. View Article : Google Scholar : PubMed/NCBI
39	Takeuchi A, Shiota M, Tatsugami K, Yokomizo A, Tanaka S, Kuroiwa K, Eto M and Naito S: p300 mediates cellular resistance to doxorubicin in bladder cancer. Mol Med Rep. 5:173–176. 2012.
40	Mees ST, Mardin WA, Wendel C, Baeumer N, Willscher E, Senninger N, Schleicher C, Colombo-Benkmann M and Haier J: EP300 - a miRNA-regulated metastasis suppressor gene in ductal adenocarcinomas of the pancreas. Int J Cancer. 126:114–124. 2010. View Article : Google Scholar
41	Callaghan R, Luk F and Bebawy M: Inhibition of the multidrug resistance P-glycoprotein: Time for a change of strategy? Drug Metab Dispos. 42:623–631. 2014. View Article : Google Scholar : PubMed/NCBI
42	Chekhun VF, Lukyanova NY, Kovalchuk O, Tryndyak VP and Pogribny IP: Epigenetic profiling of multidrug-resistant human MCF-7 breast adenocarcinoma cells reveals novel hyper- and hypomethylated targets. Mol Cancer Ther. 6:1089–1098. 2007. View Article : Google Scholar : PubMed/NCBI
43	Lu L, Zhou D, Jiang X, Song K, Li K and Ding W: Loss of E-cadherin in multidrug resistant breast cancer cell line MCF-7/Adr: Possible implication in the enhanced invasive ability. Eur Rev Med Pharmacol Sci. 16:1271–1279. 2012.PubMed/NCBI
44	Doyle L and Ross DD: Multidrug resistance mediated by the breast cancer resistance protein BCRP (ABCG2). Oncogene. 22:7340–7358. 2003. View Article : Google Scholar : PubMed/NCBI
45	Sprowl JA, Reed K, Armstrong SR, Lanner C, Guo B, Kalatskaya I, Stein L, Hembruff SL, Tam A and Parissenti AM: Alterations in tumor necrosis factor signaling pathways are associated with cytotoxicity and resistance to taxanes: A study in isogenic resistant tumor cells. Breast Cancer Res. 14:R22012. View Article : Google Scholar : PubMed/NCBI
46	Park SJ, Wu CH, Gordon JD, Zhong X, Emami A and Safa AR: Taxol induces caspase-10-dependent apoptosis. J Biol Chem. 279:51057–51067. 2004. View Article : Google Scholar : PubMed/NCBI
47	Janssen K, Pohlmann S, Jänicke RU, Schulze-Osthoff K and Fischer U: Apaf-1 and caspase-9 deficiency prevents apoptosis in a Bax-controlled pathway and promotes clonogenic survival during paclitaxel treatment. Blood. 110:3662–3672. 2007. View Article : Google Scholar : PubMed/NCBI
48	Jelínek M, Balušíková K, Kopperová D, Nĕmcová-Fürstová V, Šrámek J, Fidlerová J, Zanardi I, Ojima I and Kovář J: Caspase-2 is involved in cell death induction by taxanes in breast cancer cells. Cancer Cell Int. 13:422013. View Article : Google Scholar : PubMed/NCBI
49	Yamakawa N, Takahashi A, Mori E, Imai Y, Furusawa Y, Ohnishi K, Kirita T and Ohnishi T: High LET radiation enhances apoptosis in mutated p53 cancer cells through caspase-9 activation. Cancer Sci. 99:1455–1460. 2008. View Article : Google Scholar : PubMed/NCBI