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Autophagy inhibition re-sensitizes pulse stimulation-selected paclitaxel-resistant triple negative breast cancer cells to chemotherapy-induced apoptosis

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Abstract

Chemotherapy is the mainstay of systemic treatment for triple negative breast cancer (TNBC); however, the development of drug resistance limits its effectiveness. Therefore, we investigated the underlying mechanism for drug resistance and potential approaches to overcome it for a more effective treatment for TNBCs. Using a pulse-stimulated selection strategy to mimic chemotherapy administration in the clinic, we developed a new paclitaxel-resistant MDA-MB-231 cell line and analyzed these cells for changes in autophagy activity, and the role and mechanisms of the increased autophagy in promoting drug resistance were determined. We found that the pulse-stimulated selection strategy with paclitaxel resulted in MDA-MB-231 variant cells with enhanced resistance to paclitaxel. These resistant cells were found to have enhanced basal autophagy activity, which confers a cytoprotective function under paclitaxel treatment stress. Inhibition of autophagy enhanced paclitaxel-induced cell death in these paclitaxel-resistant cells. We further revealed that up-regulated autophagy in resistant cells enhanced the clearance of damaged mitochondria. Last, we showed that the paclitaxel-resistant cancer cells acquired cross resistance to epirubicin and cisplatin. Together, these results suggest that combining autophagy inhibition with chemotherapy may be an effective strategy to improve treatment outcome in paclitaxel-resistant TNBC patients.

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References

  1. Zeng H, Zheng R, Zhang S, Zou X, Chen W (2014) Female breast cancer statistics of 2010 in China: estimates based on data from 145 population-based cancer registries. J thorac dis 6(5):466–470

    PubMed Central  PubMed  Google Scholar 

  2. Siegel R, Ma J, Zou Z, Jemal A (2014) Cancer statistics, 2014. CA Cancer J Clin 64(1):9–29

    Article  PubMed  Google Scholar 

  3. DeSantis C, Ma J, Bryan L, Jemal A (2014) Breast cancer statistics, 2013. CA Cancer J Clin 64(1):52–62

    Article  PubMed  Google Scholar 

  4. Zagouri F, Sergentanis TN, Tsigginou A, Dimitrakakis C, Zografos GC, Dimopoulos MA, Psaltopoulou T (2014) Female breast cancer in Europe: statistics, diagnosis and treatment modalities. J thorac dis 6(6):589–590

    PubMed Central  PubMed  Google Scholar 

  5. DeSantis CE, Lin CC, Mariotto AB, Siegel RL, Stein KD, Kramer JL, Alteri R, Robbins AS, Jemal A (2014) Cancer treatment and survivorship statistics, 2014. CA Cancer J Clin 64(4):252–271

    Article  PubMed  Google Scholar 

  6. Foulkes WD, Smith IE, Reis-Filho JS (2010) Triple-negative breast cancer. New Engl J Med 363(20):1938–1948

    Article  CAS  PubMed  Google Scholar 

  7. Stevens KN, Vachon CM, Couch FJ (2013) Genetic susceptibility to triple-negative breast cancer. Cancer Res 73(7):2025–2030

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  8. Jordan MA, Wilson L (2004) Microtubules as a target for anticancer drugs. Nat Rev Cancer 4(4):253–265

    Article  CAS  PubMed  Google Scholar 

  9. Kovacs P, Csaba G, Pallinger E, Czaker R (2007) Effects of taxol treatment on the microtubular system and mitochondria of Tetrahymena. Cell Biol Int 31(7):724–732

    Article  CAS  PubMed  Google Scholar 

  10. Mizushima N (2007) Autophagy: process and function. Genes Dev 21(22):2861–2873

    Article  CAS  PubMed  Google Scholar 

  11. Wirth M, Joachim J, Tooze SA (2013) Autophagosome formation–the role of ULK1 and Beclin1-PI3KC3 complexes in setting the stage. Semin Cancer Biol 23(5):301–309

    Article  CAS  PubMed  Google Scholar 

  12. Behrends C, Sowa ME, Gygi SP, Harper JW (2010) Network organization of the human autophagy system. Nature 466(7302):68–76

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  13. Lamb CA, Yoshimori T, Tooze SA (2013) The autophagosome: origins unknown, biogenesis complex. Nat Rev Mol Cell Biol 14(12):759–774

    Article  CAS  PubMed  Google Scholar 

  14. Xie ZP, Klionsky DJ (2007) Autophagosome formation: core machinery and adaptations. Nat Cell Biol 9(10):1102–1109

    Article  CAS  PubMed  Google Scholar 

  15. Choi AM, Ryter SW, Levine B (2013) Autophagy in human health and disease. New Engl J Med 368(7):651–662

    Article  CAS  PubMed  Google Scholar 

  16. Wei H, Wei S, Gan B, Peng X, Zou W, Guan JL (2011) Suppression of autophagy by FIP200 deletion inhibits mammary tumorigenesis. Genes Dev 25(14):1510–1527

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Lock R, Roy S, Kenific CM, Su JS, Salas E, Ronen SM, Debnath J (2011) Autophagy facilitates glycolysis during Ras-mediated oncogenic transformation. Mol Biol Cell 22(2):165–178

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Sun W-L, Chen J, Wang Y-P, Zheng H (2011) Autophagy protects breast cancer cells from epirubicin-induced apoptosis and facilitates epirubicin-resistance development. Autophagy 7(9):1035–1044

    Article  CAS  PubMed  Google Scholar 

  19. Chittaranjan S, Bortnik S, Dragowska WH, Xu J, Abeysundara N, Leung A, Go NE, DeVorkin L, Weppler SA, Gelmon K et al (2014) Autophagy inhibition augments the anticancer effects of epirubicin treatment in anthracycline-sensitive and -resistant triple-negative breast cancer. Clin Cancer Res 20(12):3159–3173

    Article  CAS  PubMed  Google Scholar 

  20. Agarwal R, Kaye SB (2003) Ovarian cancer: strategies for overcoming resistance to chemotherapy. Nat Rev Cancer 3(7):502–516

    Article  CAS  PubMed  Google Scholar 

  21. Andre N, Braguer D, Brasseur G, Goncalves A, Lemesle-Meunier D, Guise S, Jordan MA, Briand C (2000) Paclitaxel induces release of cytochrome c from mitochondria isolated from human neuroblastoma cells’. Cancer Res 60(19):5349–5353

    CAS  PubMed  Google Scholar 

  22. Chen S, Jiang YZ, Huang L, Zhou RJ, Yu KD, Liu Y, Shao ZM (2013) The residual tumor autophagy marker LC3B serves as a prognostic marker in local advanced breast cancer after neoadjuvant chemotherapy. Clin Cancer Res: Off J Am Assoc Cancer Res 19(24):6853–6862

    Article  CAS  Google Scholar 

  23. Liu J, Xia H, Kim M, Xu L, Li Y, Zhang L, Cai Y, Norberg HV, Zhang T, Furuya T et al (2011) Beclin1 controls the levels of p53 by regulating the deubiquitination activity of USP10 and USP13. Cell 147(1):223–234

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  24. Wang X (2001) The expanding role of mitochondria in apoptosis. Genes Dev 15(22):2922–2933

    CAS  PubMed  Google Scholar 

  25. Gottlieb E, Armour SM, Harris MH, Thompson CB (2003) Mitochondrial membrane potential regulates matrix configuration and cytochrome c release during apoptosis. Cell Death Differ 10(6):709–717

    Article  CAS  PubMed  Google Scholar 

  26. Lee J, Giordano S, Zhang J (2012) Autophagy, mitochondria and oxidative stress: cross-talk and redox signalling. Biochem J 441(2):523–540

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Pendergrass W, Wolf N, Poot M (2004) Efficacy of MitoTracker Green and CMXrosamine to measure changes in mitochondrial membrane potentials in living cells and tissues. Cytometry Part A: J Int Soc Anal Cytol 61(2):162–169

    Article  CAS  Google Scholar 

  28. Misawa T, Takahama M, Kozaki T, Lee H, Zou J, Saitoh T, Akira S (2013) Microtubule-driven spatial arrangement of mitochondria promotes activation of the NLRP3 inflammasome. Nat Immunol 14(5):454–460

    Article  CAS  PubMed  Google Scholar 

  29. Amaravadi RK, Lippincott-Schwartz J, Yin XM, Weiss WA, Takebe N, Timmer W, DiPaola RS, Lotze MT, White E (2011) Principles and current strategies for targeting autophagy for cancer treatment. Clin Cancer Res: Off J Am Assoc Cancer Res 17(4):654–666

    Article  CAS  Google Scholar 

  30. Guo JY, Xia B, White E (2013) Autophagy-mediated tumor promotion. Cell 155(6):1216–1219

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  31. Guo JY, Chen HY, Mathew R, Fan J, Strohecker AM, Karsli-Uzunbas G, Kamphorst JJ, Chen G, Lemons JM, Karantza V et al (2011) Activated Ras requires autophagy to maintain oxidative metabolism and tumorigenesis. Genes Dev 25(5):460–470

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  32. Yang S, Wang X, Contino G, Liesa M, Sahin E, Ying H, Bause A, Li Y, Stommel JM, Dell’antonio G et al (2011) Pancreatic cancers require autophagy for tumor growth. Genes Dev 25(7):717–729

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  33. Karsli-Uzunbas G, Guo JY, Price S, Teng X, Laddha SV, Khor S, Kalaany NY, Jacks T, Chan CS, Rabinowitz JD et al (2014) Autophagy is required for glucose homeostasis and lung tumor maintenance. Cancer Discov 4(8):914–927

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  34. Hu YL, DeLay M, Jahangiri A, Molinaro AM, Rose SD, Carbonell WS, Aghi MK (2012) Hypoxia-induced autophagy promotes tumor cell survival and adaptation to antiangiogenic treatment in glioblastoma. Cancer Res 72(7):1773–1783

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  35. Strohecker AM, Guo JY, Karsli-Uzunbas G, Price SM, Chen GJ, Mathew R, McMahon M, White E (2013) Autophagy Sustains mitochondrial glutamine metabolism and growth of BrafV600E-driven lung tumors. Cancer Discov 3(11):1272–1285

    Article  CAS  PubMed  Google Scholar 

  36. Sui X, Chen R, Wang Z, Huang Z, Kong N, Zhang M, Han W, Lou F, Yang J, Zhang Q et al (2013) Autophagy and chemotherapy resistance: a promising therapeutic target for cancer treatment. Cell Death Dis 4:e838. doi:10.1038/cddis.2013.350

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  37. Amaravadi RK, Yu D, Lum JJ, Bui T, Christophorou MA, Evan GI, Thomas-Tikhonenko A, Thompson CB (2007) Autophagy inhibition enhances therapy-induced apoptosis in a Myc-induced model of lymphoma. J Clin Investig 117(2):326–336

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  38. Apel A, Herr I, Schwarz H, Rodemann HP, Mayer A (2008) Blocked autophagy sensitizes resistant carcinoma cells to radiation therapy. Cancer Res 68(5):1485–1494

    Article  CAS  PubMed  Google Scholar 

  39. Fan QW, Cheng C, Hackett C, Feldman M, Houseman BT, Nicolaides T, Haas-Kogan D, James CD, Oakes SA, Debnath J et al (2010) Akt and autophagy cooperate to promote survival of drug-resistant glioma. Sci Signal 3(147):ra81. doi:10.1126/scisignal.2001017

    PubMed Central  CAS  PubMed  Google Scholar 

  40. Huang J, Ni J, Liu K, Yu Y, Xie M, Kang R, Vernon P, Cao L, Tang D (2012) HMGB1 promotes drug resistance in osteosarcoma. Cancer Res 72(1):230–238

    Article  CAS  PubMed  Google Scholar 

  41. Levy JM, Thompson JC, Griesinger AM, Amani V, Donson AM, Birks DK, Morgan MJ, Mirsky DM, Handler MH, Foreman NK et al (2014) Autophagy inhibition improves chemosensitivity in BRAFV600E brain tumors. Cancer Discov 4(7):773–780

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  42. Liu L, Yang M, Kang R, Wang Z, Zhao Y, Yu Y, Xie M, Yin X, Livesey KM, Lotze MT et al (2011) HMGB1-induced autophagy promotes chemotherapy resistance in leukemia cells. Leukemia 25(1):23–31

    Article  PubMed  Google Scholar 

  43. Lopez G, Torres K, Liu J, Hernandez B, Young E, Belousov R, Bolshakov S, Lazar AJ, Slopis JM, McCutcheon IE et al (2011) Autophagic survival in resistance to histone deacetylase inhibitors: novel strategies to treat malignant peripheral nerve sheath tumors. Cancer Res 71(1):185–196

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  44. Nguyen HG, Yang JC, Kung HJ, Shi XB, Tilki D, Lara PN Jr, Devere White RW, Gao AC, Evans CP (2014) Targeting autophagy overcomes enzalutamide resistance in castration-resistant prostate cancer cells and improves therapeutic response in a xenograft model. Oncogene 33(36):4521–4530

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  45. O’Donovan TR, O’Sullivan GC, McKenna SL (2011) Induction of autophagy by drug-resistant esophageal cancer cells promotes their survival and recovery following treatment with chemotherapeutics. Autophagy 7(5):509–524

    Article  PubMed Central  PubMed  Google Scholar 

  46. Qadir MA, Kwok B, Dragowska WH, To KH, Le D, Bally MB, Gorski SM (2008) Macroautophagy inhibition sensitizes tamoxifen-resistant breast cancer cells and enhances mitochondrial depolarization. Breast Cancer Res Treat 112(3):389–403

    Article  CAS  PubMed  Google Scholar 

  47. Song J, Qu Z, Guo X, Zhao Q, Zhao X, Gao L, Sun K, Shen F, Wu M, Wei L (2009) Hypoxia-induced autophagy contributes to the chemoresistance of hepatocellular carcinoma cells. Autophagy 5(8):1131–1144

    Article  CAS  PubMed  Google Scholar 

  48. Zhao H, Yang M, Zhao J, Wang J, Zhang Y, Zhang Q (2013) High expression of LC3B is associated with progression and poor outcome in triple-negative breast cancer. Med Oncol 30(1):475

    Article  PubMed  Google Scholar 

  49. Choi J, Jung W, Koo JS (2013) Expression of autophagy-related markers beclin-1, light chain 3A, light chain 3B and p62 according to the molecular subtype of breast cancer. Histopathology 62(2):275–286

    Article  PubMed  Google Scholar 

  50. Ladoire S, Chaba K, Martins I, Sukkurwala AQ, Adjemian S, Michaud M, Poirier-Colame V, Andreiuolo F, Galluzzi L, White E et al (2012) Immunohistochemical detection of cytoplasmic LC3 puncta in human cancer specimens. Autophagy 8(8):1175–1184

    Article  PubMed Central  PubMed  Google Scholar 

  51. Samaddar JS, Gaddy VT, Duplantier J, Thandavan SP, Shah M, Smith MJ, Browning D, Rawson J, Smith SB, Barrett JT et al (2008) A role for macroautophagy in protection against 4-hydroxytamoxifen-induced cell death and the development of antiestrogen resistance. Mol Cancer Ther 7(9):2977–2987

    Article  CAS  PubMed  Google Scholar 

  52. Vazquez-Martin A, Oliveras-Ferraros C, Menendez JA (2009) Autophagy facilitates the development of breast cancer resistance to the anti-HER2 monoclonal antibody trastuzumab. PLoS One 4(7):e6251

    Article  PubMed Central  PubMed  Google Scholar 

  53. Maycotte P, Gearheart CM, Barnard R, Aryal S, Mulcahy Levy JM, Fosmire SP, Hansen RJ, Morgan MJ, Porter CC, Gustafson DL et al (2014) STAT3-mediated autophagy dependence identifies subtypes of breast cancer where autophagy inhibition can be efficacious. Cancer Res 74(9):2579–2590

    Article  CAS  PubMed  Google Scholar 

  54. Varbiro G, Veres B, Gallyas F Jr, Sumegi B (2001) Direct effect of Taxol on free radical formation and mitochondrial permeability transition. Free Radic Biol Med 31(4):548–558

    Article  CAS  PubMed  Google Scholar 

  55. Fulda S, Galluzzi L, Kroemer G (2010) Targeting mitochondria for cancer therapy. Nat Rev Drug Discov 9(6):447–464

    Article  CAS  PubMed  Google Scholar 

  56. Namba T, Takabatake Y, Kimura T, Takahashi A, Yamamoto T, Matsuda J, Kitamura H, Niimura F, Matsusaka T, Iwatani H et al (2014) Autophagic clearance of mitochondria in the kidney copes with metabolic acidosis. J Am Soc Nephrol 25(10):2254–2266

    Article  CAS  PubMed  Google Scholar 

  57. Randow F, Youle RJ (2014) Self and nonself: how autophagy targets mitochondria and bacteria. Cell Host Microbe 15(4):403–411

    Article  CAS  PubMed  Google Scholar 

  58. Boyer-Guittaut M, Poillet L, Liang Q, Bole-Richard E, Ouyang X, Benavides GA, Chakrama FZ, Fraichard A, Darley-Usmar VM, Despouy G et al (2014) The role of GABARAPL1/GEC1 in autophagic flux and mitochondrial quality control in MDA-MB-436 breast cancer cells. Autophagy 10(6):986–1003

    Article  CAS  PubMed  Google Scholar 

  59. Wind NS, Holen I (2011) Multidrug resistance in breast cancer: from in vitro models to clinical studies. Int J Breast Cancer 2011 (967419). 10.4061/2011/967419

  60. Wang J, Wu GS (2014) Role of autophagy in cisplatin resistance in ovarian cancer cells. J Biol Chem 289(24):17163–17173

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This research was supported by NIH Grant CA163493 and CA150926 to J.-L. Guan.

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The authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, OH, 45267, USA

    Jian Wen, Syn Yeo, Chenran Wang, Song Chen, Shaogang Sun, Michael A. Haas & Jun-Lin Guan

  2. Department of Breast Surgery, The Fourth Affiliated Hospital of China Medical University, Shenyang, 110032, Liaoning, China

    Jian Wen & Wei Tu

  3. Department of Breast Surgery, The First Affiliated Hospital of China Medical University, Shenyang, 110001, Liaoning, China

    Jian Wen & Feng Jin

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  1. Jian Wen
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  2. Syn Yeo
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  4. Song Chen
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  9. Jun-Lin Guan
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Correspondence to Jun-Lin Guan.

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Wen, J., Yeo, S., Wang, C. et al. Autophagy inhibition re-sensitizes pulse stimulation-selected paclitaxel-resistant triple negative breast cancer cells to chemotherapy-induced apoptosis. Breast Cancer Res Treat 149, 619–629 (2015). https://doi.org/10.1007/s10549-015-3283-9

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