Synergism of a novel MCL‑1 downregulator, acriflavine, with navitoclax (ABT‑263) in triple‑negative breast cancer, lung adenocarcinoma and glioblastoma multiforme

Lee,Anbok; Jin,Hyeon-Ok; Masudul Haque,Md.; Kim,Hee Yeon; Jung,Hana; Park,Jin Hee; Kim,Ilhwan; Song,Joo Yeon; Yoon,Hye Kyoung; Kim,Hyoung Kyu; Han,Jin; Park,In-Chul; Kim,Kwang Seok; Park,Sae Gwang

doi:10.3892/ijo.2021.5292

Synergism of a novel MCL‑1 downregulator, acriflavine, with navitoclax (ABT‑263) in triple‑negative breast cancer, lung adenocarcinoma and glioblastoma multiforme

Authors:
- Anbok Lee
- Hyeon-Ok Jin
- Md. Masudul Haque
- Hee Yeon Kim
- Hana Jung
- Jin Hee Park
- Ilhwan Kim
- Joo Yeon Song
- Hye Kyoung Yoon
- Hyoung Kyu Kim
- Jin Han
- In-Chul Park
- Kwang Seok Kim
- Sae Gwang Park
View Affiliations

Affiliations: Department of Surgery, Busan Paik Hospital, College of Medicine, Inje University, Busan 47392, Republic of Korea, KIRAMS Radiation Biobank, Korea Institute of Radiological and Medical Sciences, Seoul 01812, Republic of Korea, Department of Microbiology and Immunology, College of Medicine, Inje University, Busan 47392, Republic of Korea, Department of Internal Medicine, Division of Oncology, Haeundae Paik Hospital, College of Medicine, Inje University, Busan 48108, Republic of Korea, Department of Pathology, Dongnam Institute of Radiological and Medical Sciences, Busan 46033, Republic of Korea, Department of Pathology, Inje University, Busan 47392, Republic of Korea, Cardiovascular and Metabolic Disease Center, Smart Marine Therapeutic Center, College of Medicine, Inje University, Busan 47397, Republic of Korea, Division of Radiation Biomedical Research, Korea Institute of Radiological and Medical Sciences, Seoul 01812, Republic of Korea
Published online on: December 8, 2021 https://doi.org/10.3892/ijo.2021.5292
Article Number: 2
Copyright: © Lee et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Myeloid cell leukemia sequence 1 (MCL‑1), an anti‑apoptotic B‑cell lymphoma 2 (BCL‑2) family molecule frequently amplified in various human cancer cells, is known to be critical for cancer cell survival. MCL‑1 has been recognized as a target molecule for cancer treatment. While various agents have emerged as potential MCL‑1 blockers, the present study presented acriflavine (ACF) as a novel MCL‑1 inhibitor in triple‑negative breast cancer (TNBC). Further evaluation of its treatment potential on lung adenocarcinoma and glioblastoma multiforme (GBM) was also investigated. The anticancer effect of ACF on TNBC cells was demonstrated when MDA‑MB‑231 and HS578T cells were treated with ACF. ACF significantly induced typical intrinsic apoptosis in TNBCs in a dose‑ and time‑dependent manner via MCL‑1 downregulation. MCL‑1 downregulation by ACF treatment was revealed at each phase of protein expression. Initially, transcriptional regulation via reverse transcription‑quantitative PCR was validated. Then, post‑translational regulation was explained by utilizing an inhibitor against protein biosynthesis and proteasome. Lastly, immunoprecipitation of ubiquitinated MCL‑1 confirmed the post‑translational downregulation of MCL‑1. In addition, the synergistic treatment efficacy of ACF with the well‑known MCL‑1 inhibitor ABT‑263 against the TNBC cells was explored [combination index (CI)<1]. Conjointly, the anticancer effect of ACF was assessed in GBM (U87, U251 and U343), and lung cancer (A549 and NCI‑H69) cell lines as well, using immunoblotting, cytotoxicity assay and FACS. The effect of the combination treatment using ACF and ABT‑263 was estimated in GBM (U87, U343 and U251), and non‑small cell lung cancer (A549) cells likewise. The present study suggested a novel MCL‑1 inhibitory function of ACF and the synergistic antitumor effect with ABT‑263.

View Figures

View References

1	Foulkes WD, Smith IE and Reis-Filho JS: Triple-negative breast cancer. N Engl J Med. 363:1938–1948. 2010. View Article : Google Scholar : PubMed/NCBI
2	Adams JM and Cory S: The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene. 26:1324–1337. 2007. View Article : Google Scholar : PubMed/NCBI
3	Hanahan D and Weinberg RA: Hallmarks of cancer: The next generation. Cell. 144:646–674. 2011. View Article : Google Scholar : PubMed/NCBI
4	Adams JM and Cory S: The BCL-2 arbiters of apoptosis and their growing role as cancer targets. Cell Death Differ. 25:27–36. 2018. View Article : Google Scholar
5	Mohammad RM, Muqbil I, Lowe L, Yedjou C, Hsu HY, Lin LT, Siegelin MD, Fimognari C, Kumar NB, Dou QP, et al: Broad targeting of resistance to apoptosis in cancer. Semin Cancer Biol. 35(Suppl): S78–S103. 2015. View Article : Google Scholar : PubMed/NCBI
6	Kang MH and Reynolds CP: Bcl-2 inhibitors: Targeting mitochondrial apoptotic pathways in cancer therapy. Clin Cancer Res. 15:1126–1132. 2009. View Article : Google Scholar : PubMed/NCBI
7	Ertel F, Nguyen M, Roulston A and Shore GC: Programming cancer cells for high expression levels of Mcl1. EMBO Rep. 14:328–336. 2013. View Article : Google Scholar : PubMed/NCBI
8	Craig RW: MCL1 provides a window on the role of the BCL2 family in cell proliferation, differentiation and tumorigenesis. Leukemia. 16:444–454. 2002. View Article : Google Scholar : PubMed/NCBI
9	Beroukhim R, Mermel CH, Porter D, Wei G, Raychaudhuri S, Donovan J, Barretina J, Boehm JS, Dobson J, Urashima M, et al: The landscape of somatic copy-number alteration across human cancers. Nature. 463:899–905. 2010. View Article : Google Scholar : PubMed/NCBI
10	Ding Q, He X, Hsu JM, Xia W, Chen CT, Li LY, Lee DF, Liu JC, Zhong Q, Wang X and Hung MC: Degradation of Mcl-1 by beta-TrCP mediates glycogen synthase kinase 3-induced tumor suppression and chemosensitization. Mol Cell Biol. 27:4006–4017. 2007. View Article : Google Scholar : PubMed/NCBI
11	Young AI, Timpson P, Gallego-Ortega D, Ormandy CJ and Oakes SR: Myeloid cell leukemia 1 (MCL-1), an unexpected modulator of protein kinase signaling during invasion. Cell Adh Migr. 12:513–523. 2018. View Article : Google Scholar
12	Woo SM, Min KJ, Seo BR, Seo YH, Jeong YJ and Kwon TK: YM155 enhances ABT-737-mediated apoptosis through Mcl-1 downregulation in Mcl-1-overexpressed cancer cells. Mol Cell Biochem. 429:91–102. 2017. View Article : Google Scholar : PubMed/NCBI
13	Caenepeel S, Brown SP, Belmontes B, Moody G, Keegan KS, Chui D, Whittington DA, Huang X, Poppe L, Cheng AC, et al: AMG 176, a selective MCL1 inhibitor, is effective in hematologic cancer models alone and in combination with established therapies. Cancer Discov. 8:1582–1597. 2018.PubMed/NCBI
14	Ramsey HE, Fischer MA, Lee T, Gorska AE, Arrate MP, Fuller L, Boyd KL, Strickland SA, Sensintaffar J, Hogdal LJ, et al: A novel MCL1 inhibitor combined with venetoclax rescues venetoclaxresistant acute myelogenous leukemia. Cancer Discov. 8:1566–1581. 2018. View Article : Google Scholar : PubMed/NCBI
15	Wainwright M: Acridine-a neglected antibacterial chromophore. J Antimicrob Chemother. 47:1–13. 2001. View Article : Google Scholar : PubMed/NCBI
16	Lee K, Zhang H, Qian DZ, Rey S, Liu JO and Semenza GL: Acriflavine inhibits HIF-1 dimerization, tumor growth, and vascularization. Proc Natl Acad Sci USA. 106:17910–17915. 2009. View Article : Google Scholar : PubMed/NCBI
17	Zhang L, Huang G, Li X, Zhang Y, Jiang Y, Shen J, Liu J, Wang Q, Zhu J, Feng X, et al: Hypoxia induces epithelial-mesenchymal transition via activation of SNAI1 by hypoxia-inducible factor -1α in hepatocellular carcinoma. BMC Cancer. 13:1082013. View Article : Google Scholar
18	Lee CJ, Yue CH, Lin YJ, Lin YY, Kao SH, Liu JY and Chen YH: Antitumor activity of acriflavine in lung adenocarcinoma cell line A549. Anticancer Res. 34:6467–6472. 2014.PubMed/NCBI
19	Lee CJ, Yue CH, Lin YY, Wu JC and Liu JY: Antitumor activity of acriflavine in human hepatocellular carcinoma cells. Anticancer Res. 34:3549–3556. 2014.PubMed/NCBI
20	Dekervel J, Bulle A, Windmolders P, Lambrechts D, Van Cutsem E, Verslype C and van Pelt J: Acriflavine inhibits acquired drug resistance by blocking the epithelial-to-mesenchymal transition and the unfolded protein response. Transl Oncol. 10:59–69. 2017. View Article : Google Scholar :
21	Mangraviti A, Raghavan T, Volpin F, Skuli N, Gullotti D, Zhou J, Asnaghi L, Sankey E, Liu A, Wang Y, et al: HIF-1α-targeting acriflavine provides long term survival and radiological tumor response in brain cancer therapy. Sci Rep. 7:149782017. View Article : Google Scholar
22	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
23	Jeong SH, Kim HK, Song IS, Lee SJ, Ko KS, Rhee BD, Kim N, Mishchenko NP, Fedoryev SA, Stonik VA and Han J: Echinochrome A protects mitochondrial function in cardiomyocytes against cardiotoxic drugs. Mar Drugs. 12:2922–2936. 2014. View Article : Google Scholar : PubMed/NCBI
24	Chou TC: Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev. 58:621–681. 2006. View Article : Google Scholar : PubMed/NCBI
25	Ashton JC: Drug combination studies and their synergy quantification using the Chou-Talalay method-letter. Cancer Res. 75:24002015. View Article : Google Scholar
26	Williams MM, Lee L, Hicks DJ, Joly MM, Elion D, Rahman B, McKernan C, Sanchez V, Balko JM, Stricker T, et al: Key survival factor, Mcl-1, correlates with sensitivity to combined Bcl-2/Bcl-xL blockade. Mol Cancer Res. 15:259–268. 2017. View Article : Google Scholar : PubMed/NCBI
27	Certo M, Del Gaizo Moore V, Nishino M, Wei G, Korsmeyer S, Armstrong SA and Letai A: Mitochondria primed by death signals determine cellular addiction to antiapoptotic BCL-2 family members. Cancer Cell. 9:351–365. 2006. View Article : Google Scholar : PubMed/NCBI
28	Lovell JF, Billen LP, Bindner S, Shamas-Din A, Fradin C, Leber B and Andrews DW: Membrane binding by tBid initiates an ordered series of events culminating in membrane permeabilization by Bax. Cell. 135:1074–1084. 2008. View Article : Google Scholar : PubMed/NCBI
29	Kuwana T, Bouchier-Hayes L, Chipuk JE, Bonzon C, Sullivan BA, Green DR and Newmeyer DD: BH3 domains of BH3-only proteins differentially regulate Bax-mediated mitochondrial membrane permeabilization both directly and indirectly. Mol Cell. 17:525–535. 2005. View Article : Google Scholar : PubMed/NCBI
30	Willis SN, Chen L, Dewson G, Wei A, Naik E, Fletcher JI, Adams JM and Huang DC: Proapoptotic bak is sequestered by Mcl-1 and Bcl-xL, but not Bcl-2, until displaced by BH3-only proteins. Genes Dev. 19:1294–1305. 2005. View Article : Google Scholar : PubMed/NCBI
31	Edlich F, Banerjee S, Suzuki M, Cleland MM, Arnoult D, Wang C, Neutzner A, Tjandra N and Youle RJ: Bcl-x(L) retrotranslocates bax from the mitochondria into the cytosol. Cell. 145:104–116. 2011. View Article : Google Scholar : PubMed/NCBI
32	Oltersdorf T, Elmore SW, Shoemaker AR, Armstrong RC, Augeri DJ, Belli BA, Bruncko M, Deckwerth TL, Dinges J, Hajduk PJ, et al: An inhibitor of Bcl-2 family proteins induces regression of solid tumours. Nature. 435:677–681. 2005. View Article : Google Scholar : PubMed/NCBI
33	Souers AJ, Leverson JD, Boghaert ER, Ackler SL, Catron ND, Chen J, Dayton BD, Ding H, Enschede SH, Fairbrother WJ, et al: ABT-199, a potent and selective BCL-2 inhibitor, achieves anti-tumor activity while sparing platelets. Nat Med. 19:202–208. 2013. View Article : Google Scholar : PubMed/NCBI
34	Bartlett NL: Highlights in lymphoma from the 2013 American society of hematology annual meeting and exposition: Commentary. Clin Adv Hematol Oncol. 12(Suppl 6): 18–23. 2014.PubMed/NCBI
35	Sieghart W, Losert D, Strommer S, Cejka D, Schmid K, Rasoul-Rockenschaub S, Bodingbauer M, Crevenna R, Monia BP, Peck-Radosavljevic M and Wacheck V: Mcl-1 overexpression in hepatocellular carcinoma: A potential target for antisense therapy. J Hepatol. 44:151–157. 2006. View Article : Google Scholar
36	LaBelle JL, Katz SG, Bird GH, Gavathiotis E, Stewart ML, Lawrence C, Fisher JK, Godes M, Pitter K, Kung AL and Walensky LD: A stapled BIM peptide overcomes apoptotic resistance in hematologic cancers. J Clin Invest. 122:2018–2031. 2012. View Article : Google Scholar : PubMed/NCBI
37	Reiner T, de Las Pozas A, Parrondo R, Palenzuela D, Cayuso W, Rai P and Perez-Stable C: Mcl-1 protects prostate cancer cells from cell death mediated by chemotherapy-induced DNA damage. Oncoscience. 2:703–715. 2015. View Article : Google Scholar : PubMed/NCBI
38	Young AI, Law AM, Castillo L, Chong S, Cullen HD, Koehler M, Herzog S, Brummer T, Lee EF, Fairlie WD, et al: MCL-1 inhibition provides a new way to suppress breast cancer metastasis and increase sensitivity to dasatinib. Breast Cancer Res. 18:1252016. View Article : Google Scholar : PubMed/NCBI
39	Zervantonakis IK, Iavarone C, Chen HY, Selfors LM, Palakurthi S, Liu JF, Drapkin R, Matulonis U, Leverson JD, Sampath D, et al: Systems analysis of apoptotic priming in ovarian cancer identifies vulnerabilities and predictors of drug response. Nat Commun. 8:3652017. View Article : Google Scholar : PubMed/NCBI
40	Campbell KJ, Dhayade S, Ferrari N, Sims AH, Johnson E, Mason SM, Dickson A, Ryan KM, Kalna G, Edwards J, et al: MCL-1 is a prognostic indicator and drug target in breast cancer. Cell Death Dis. 9:192018. View Article : Google Scholar : PubMed/NCBI
41	Goodwin CM, Rossanese OW, Olejniczak ET and Fesik SW: Myeloid cell leukemia-1 is an important apoptotic survival factor in triple-negative breast cancer. Cell Death Differ. 22:2098–2106. 2015. View Article : Google Scholar : PubMed/NCBI
42	Balko JM, Giltnane JM, Wang K, Schwarz LJ, Young CD, Cook RS, Owens P, Sanders ME, Kuba MG, Sánchez V, et al: Molecular profiling of the residual disease of triple-negative breast cancers after neoadjuvant chemotherapy identifies actionable therapeutic targets. Cancer Discov. 4:232–245. 2014. View Article : Google Scholar :
43	Ozretic P, Alvir I, Sarcevic B, Vujaskovic Z, Rendic-Miocevic Z, Roguljic A and Beketic-Oreskovic L: Apoptosis regulator Bcl-2 is an independent prognostic marker for worse overall survival in triple-negative breast cancer patients. Int J Biol Markers. 33:109–115. 2018. View Article : Google Scholar
44	Inuzuka H, Shaik S, Onoyama I, Gao D, Tseng A, Maser RS, Zhai B, Wan L, Gutierrez A, Lau AW, et al: SCF(FBW7) regulates cellular apoptosis by targeting MCL1 for ubiquitylation and destruction. Nature. 471:104–109. 2011. View Article : Google Scholar : PubMed/NCBI
45	Wertz IE, Kusam S, Lam C, Okamoto T, Sandoval W, Anderson DJ, Helgason E, Ernst JA, Eby M, Liu J, et al: Sensitivity to antitubulin chemotherapeutics is regulated by MCL1 and FBW7. Nature. 471:110–114. 2011. View Article : Google Scholar : PubMed/NCBI
46	Leverson JD, Zhang H, Chen J, Tahir SK, Phillips DC, Xue J, Nimmer P, Jin S, Smith M, Xiao Y, et al: Potent and selective small-molecule MCL-1 inhibitors demonstrate on-target cancer cell killing activity as single agents and in combination with ABT-263 (navitoclax). Cell Death Dis. 6:e15902015. View Article : Google Scholar : PubMed/NCBI
47	Pan R, Ruvolo VR, Wei J, Konopleva M, Reed JC, Pellecchia M, Andreeff M and Ruvolo PP: Inhibition of Mcl-1 with the pan-Bcl-2 family inhibitor (-)BI97D6 overcomes ABT-737 resistance in acute myeloid leukemia. Blood. 126:363–372. 2015. View Article : Google Scholar : PubMed/NCBI
48	Mojsa B, Lassot I and Desagher S: Mcl-1 ubiquitination: Unique regulation of an essential survival protein. Cells. 3:418–437. 2014. View Article : Google Scholar : PubMed/NCBI
49	Goldie H, Walker M, Graham T and Williams F: Topical effect of acriflavine compounds on growth and spread of malignant cells. J Natl Cancer Inst. 23:841–855. 1959.PubMed/NCBI
50	Hassan S, Laryea D, Mahteme H, Felth J, Fryknas M, Fayad W, Linder S, Rickardson L, Gullbo J, Graf W, et al: Novel activity of acriflavine against colorectal cancer tumor cells. Cancer Sci. 102:2206–2213. 2011. View Article : Google Scholar : PubMed/NCBI
51	Kim SG, Kim CW, Ahn ET, Lee KY, Hong EK, Yoo BI and Han YB: Enhanced anti-tumour effects of acriflavine in combination with guanosine in mice. J Pharm Pharmacol. 49:216–222. 1997. View Article : Google Scholar : PubMed/NCBI