Single Agent and Synergistic Activity of Maritoclax with ABT-263 in Nasopharyngeal Carcinoma (NPC) Cell Lines

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Benedict Lian Shi Xiang
Lo Kwok-Wai
Alan Khoo Soo-Beng
Nethia MohanaKumaran

Abstract


The BCL-2 anti-apoptotic proteins are over-expressed in many cancers and hence are attractive therapeutic targets. In this study, we tested the sensitivity of two Nasopharyngeal Carcinoma (NPC) cell lines HK1 and C666-1 to Maritoclax, which is reported to repress anti-apoptotic protein MCL-1 and BH3 mimetic ABT-263, which selectively inhibits anti-apoptotic proteins BCL-2, BCL-XL and BCL-w. We investigated the sensitisation of the NPC cell lines to these drugs using the SYBR Green I assay and 3D NPC spheroids. We report that Maritoclax repressed anti-apoptotic proteins MCL-1, BCL-2, and BCL-XL in a dose- and time-dependent manner and displayed a single agent activity in inhibiting cell proliferation of the NPC cell lines. Moreover, combination of Maritoclax and ABT-263 exhibited synergistic antiproliferative effect in the HK1 cells. Similar results were obtained in the 3D spheroids generated from the HK1 cells. More notably, 3D HK1 spheroids either treated with single agent Maritoclax or combination with ABT-263, over 10 days, did not develop resistance to the treatment rapidly. Collectively, the findings illustrate that Maritoclax as a single agent or combination with BH3 mimetics could be potentially useful as treatment strategies for the management of NPC.


 


Protein anti-apoptotik BCL-2 diekspres pada tahap yang tinggi dalam kebanyakan kanser dan merupakan sasaran terapeutik kanser yang menarik. Dalam kajian ini, sensitiviti dua titisan sel kanser nasofarinks diuji, iaitu HK1 dan C666-1 kepada Maritoclax yang dilaporkan menurunkan tahap protein anti-apoptotik MCL-1 dan BH3 mimetik ABT-263 yang merencat protein anti-apoptotik BCL-2, BCL-XL dan BCL-w. Sensitiviti sel kanser nasofarinks kepada dadah ini disiasat menggunakan ujian SYBR Green I dan sferoid 3D kanser nasofarink. Di sini kami melaporkan bahawa Maritoclax mengurangkan tahap MCL-1, BCL-2 dan BCL-XL mengikut konsentrasi drug dan tempoh rawatan dan Maritoclax memaparkan aktiviti agen tunggal dalam menghalang percambahan sel kanser nasofarinks. Selain itu, gabungan Maritoclax dan ABT-263 menghalang percambahan sel kanser nasofarinks HK1 secara sinergistik. Keputusan yang sama diperolehi dengan sferoid 3D sel HK1. Lebih penting, sferoid 3D sel HK1, sama ada dirawat dengan agen tunggal Maritoclax atau gabungan dengan ABT-263 selama 10 hari, tidak menunjukkan resistan terhadap rawatan dengan cepat. Secara kolektif, kajian ini menunjukan bahawa Maritoclax sebagai ejen tunggal atau gabungan dengan BH3 mimetik boleh menjadi strategi rawatan yang berpotensi untuk NPC.


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How to Cite
Single Agent and Synergistic Activity of Maritoclax with ABT-263 in Nasopharyngeal Carcinoma (NPC) Cell Lines. (2020). Tropical Life Sciences Research, 31(3), 1–13. https://doi.org/10.21315/tlsr2020.31.3.1
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Original Article

References

Airiau K, Prouzet-Mauleon V, Rousseau B, Pigneux A, Jeanneteau M, Giraudon M, Allou K, Dubus P, Belloc F and Mahon F X. (2016). Synergistic cooperation between ABT-263 and MEK1/2 inhibitor: effect on apoptosis and proliferation of acute myeloid leukemia cells. Oncotarget 7(1): 845–859. https://doi.org/10.18632/oncotarget.6417

Allen J D, Jackson S C and Schinkel A H. (2002). A mutation hot spot in the Bcrp1 (Abcg2) multidrug transporter in mouse cell lines selected for Doxorubicin resistance. Cancer Research 62(8): 2294–2299.

Bray F, Ferlay J, Soerjomataram I, Siegel R L, Torre L A and Jemal A. (2018). Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians 68(6): 394–424. https://doi.org/10.3322/caac.21492

Chen M K, Lai J C, Chang C C, Chang J H, Chang Y J and Chen H C. (2008). Prognostic impact of bcl-2 expression on advanced nasopharyngeal carcinoma. Head Neck 30(8): 1052–1057. https://doi.org/10.1002/hed.20839

Chen M K, Yang S F, Lai J C, Yeh K T, Yang J S, Chen L S and Chen H C. (2010). Expression of bcl-2 correlates with poor prognosis and modulates migration of nasopharyngeal carcinoma cells. Clinica Chimica Acta 411(5–6): 400–405. https://doi.org/10.1016/j.cca.2009.12.010

Chou T C. (2006). Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacological Reviews 58(3): 621–681. https://doi.org/10.1124/pr.58.3.10

Chou T C and Talalay P. (1984). Quantitative analysis of dose-effect relationships: The combined effects of multiple drugs or enzyme inhibitors. Advances in Enzyme Regulation 22: 27–55.

Daker M, Ahmad M and Khoo A S. (2012). Quercetin-induced inhibition and synergistic activity with cisplatin: A chemotherapeutic strategy for nasopharyngeal carcinoma cells. Cancer Cell International 12(1): 34. https://doi.org/10.1186/1475-2867-12-34

Doi K, Li R, Sung S S, Wu H, Liu Y, Manieri W, Krishnegowda G et al. (2012). Discovery of marinopyrrole A (maritoclax) as a selective Mcl-1 antagonist that overcomes ABT-737 resistance by binding to and targeting Mcl-1 for proteasomal degradation. Journal of Biological Chemistry 287(13): 10224–10235. https://doi.org/10.1074/jbc.M111.334532

Doi K, Liu Q, Gowda K, Barth B M, Claxton D, Amin S, Loughran Jr. T P and Wang H G. (2014). Maritoclax induces apoptosis in acute myeloid leukemia cells with elevated Mcl-1 expression. Cancer Biology & Therapy 15(8): 1077–1086. https://doi.org/10.4161/cbt.29186

Eichhorn J M, Alford S E, Hughes C C, Fenical W and Chambers T C. (2013). Purported Mcl1 inhibitor marinopyrrole A fails to show selective cytotoxicity for Mcl-1-dependent cell lines. Cell Death & Disease 4: e880. https://doi.org/10.1038/cddis.2013.411

Fan S Q, Ma J, Zhou J, Xiong W, Xiao B Y, Zhang W L, Tan C et al. (2006). Differential expression of Epstein-Barr virus-encoded RNA and several tumor-related genes in various types of nasopharyngeal epithelial lesions and nasopharyngeal carcinoma using tissue microarray analysis. Human Pathology 37(5): 593–605. https://doi.org/10.1016/j.humpath.2006.01.010

Hughes C C, Prieto-Davo A, Jensen P R and Fenical W. (2008). The marinopyrroles, antibiotics of an unprecedented structure class from a marine Streptomyces sp. Organic Letters 10: 629–631.

Hughes C C, Yang Y L, Liu W T, Dorrestein P C, La Clair J J and Fenical W. (2009). Marinopyrrole A target elucidation by acyl dye transfer. Journal of the American Chemical Society 131: 12094–12096.

Jane E P, Premkumar D R, Cavaleri J M, Sutera P A, Rajasekar T and Pollack I F. (2016). Dinaciclib, a cyclin-dependent kinase inhibitor promotes proteasomal degradation of Mcl-1 and enhances ABT-737-mediated cell death in malignant human glioma cell lines. Journal of Pharmacology and Experimental Therapeutics 356(2): 354–365. https://doi.org/10.1124/jpet.115.230052

Leverson J D, Phillips D C, Mitten M J, Boghaert E R, Diaz D, Tahir S K, Belmont L D et al. (2015). Exploiting selective BCL-2 family inhibitors to dissect cell survival dependencies and define improved strategies for cancer therapy. Science Translational Medicine 7(279): 279ra240. https://doi.org/10.1126/scitranslmed.aaa4642

Lian B S X, Yek A E H, Shuvas H, Abdul Rahman S F, Muniandy K. and Mohana-Kumaran N. (2018). Synergistic anti-proliferative effects of combination of ABT-263 and MCL-1 selective inhibitor A-1210477 on cervical cancer cell lines. BMC Research Notes 11(1): 197. https://doi.org/10.1186/s13104-018-3302-0

Lucas K M, Mohana-Kumaran N, Lau D, Zhang X D, Hersey P, Huang D C, Weninger W, Haass N K and Allen J D. (2012). Modulation of NOXA and MCL-1 as a strategy for sensitizing melanoma cells to the BH3-mimetic ABT-737. Clinical Cancer Research 18(3): 783–795. https://doi.org/10.1158/1078-0432.CCR-11-1166

McGowan E M, Alling N, Jackson E A, Yagoub D, Haass N K, Allen J D and MartinelloWilks R. (2011). Evaluation of cell cycle arrest in estrogen responsive MCF7 breast cancer cells: Pitfalls of the MTS assay. PLoS One 6(6): e20623. https://doi.org/10.1371/journal.pone.0020623

Montero J and Letai A. (2018). Why do BCL-2 inhibitors work and where should we use them in the clinic? Cell Death and Differentiaton 25(1): 56–64. https://doi.org/10.1038/cdd.2017.183

Nakajima W, Sharma K, Hicks M A, Le N, Brown R, Krystal G W and Harada H. (2016). Combination with vorinostat overcomes ABT-263 (navitoclax) resistance of small cell lung cancer. Cancer Biology & Therapy 17(1): 27–35. https://doi.org/10.1080/15384047.2015.1108485

Pandey M K, Gowda K, Doi K, Sharma A K, Wang H G and Amin S. (2013). Proteasomal degradation of Mcl-1 by maritoclax induces apoptosis and enhances the efficacy of ABT-737 in melanoma cells. PLoS One 8(11): e78570. https://doi.org/10.1371/journal.pone.0078570

Sheu L F, Chen A, Meng C L, Ho K C, Lin F G and Lee W H. (1997). Analysis of bcl2 expression in normal, inflamed, dysplastic nasopharyngeal epithelia, and nasopharyngeal carcinoma: Association with p53 expression. Human Pathology 28(5): 556–562.

Smalley K S, Haass N K, Brafford P A, Lioni M, Flaherty K T and Herlyn M. (2006). Multiple signaling pathways must be targeted to overcome drug resistance in cell lines derived from melanoma metastases. Molecular Cancer Therapeutics 5(5): 1136–1144. https://doi.org/10.1158/1535-7163.MCT-06-0084

Smalley K S, Lioni M, Noma K, Haass N K and Herlyn M. (2008). In vitro three-dimensional tumor microenvironment models for anticancer drug discovery. Expert Opinion on Drug Discovery 3(1): 1–10. https://doi.org/10.1517/17460441.3.1.1

Tse C, Shoemaker A R, Adickes J, Anderson M G, Chen J, Jin S, Johnson E F et al. (2008). ABT-263: A potent and orally bioavailable Bcl-2 family inhibitor. Cancer Research 68(9): 3421–3428. https://doi.org/10.1158/0008-5472.CAN-07-5836

Varadarajan S, Poornima P, Milani M, Gowda K, Amin S, Wang H G and Cohen G M. (2015). Maritoclax and dinaciclib inhibit MCL-1 activity and induce apoptosis in both a MCL-1-dependent and-independent manner. Oncotarget 6(14): 12668–12681. https://doi.org/10.18632/oncotarget.3706

Yu Y, Dong W, Li X, Yu E, Zhou X and Li S. (2003). Significance of c-Myc and Bcl-2 protein expression in nasopharyngeal carcinoma. Archives of Otolaryngology–Head & Neck Surgery 129(12): 1322–1326. https://doi.org/10.1001/archotol.129.12.1322