Biopolym. Cell. 2016; 32(3):190-202.
http://dx.doi.org/10.7124/bc.000920
Genomics, Transcriptomics and Proteomics
Mitochondria do not play a major role in landomycin E-induced ROS burst and circumvention of multiple drug resistance in HL-60 leukemia cells
1Panchuk R. R., 1Lehka L. V., 2Rohr J., 3Berger W., 1Stoika R. S.
  1. Institute of Cell Biology, NAS of Ukraine
    14/16, Drahomanov Str., Lviv, Ukraine, 79005
  2. University of Kentucky, College of Pharmacy
    789 S. Limestone Str. Lexington, USA
  3. Institute of Cancer Research and Comprehensive Cancer Center, Medical University Vienna
    8a, Borschkegasse, Vienna 1090, Austria

Abstract

Aim. To study the molecular mechanisms of reactive oxygen species (ROS) involvement in circumventing the cancer drug resistance by novel angucycline antibiotic landomycin E in HL-60 human leukemia cells and its drug-resistant sublines HL-60/adr and HL-60/vinc. Methods. MTT assay, trypan blue exclusion test, DCFDA and JC-1 staining of cells. Results. Landomycin E (LE) leads to a massive hydrogen peroxide production in HL-60 cell line already 1h after the drug addition to the cell culture, while depolarization of mitochondria is observed only at 6–12h, which indicates on the extra-mitochondrial ROS production by LE. The drug-resistant cells of HL-60/vinc (P-gp+) despite 100-fold resistance to doxorubicin (Dx) action, demonstrated no difference in the resistance to LE compared to the parental cell line, while HL-60/adr line (MRP-1+), which was found to be 200-fold resistant to Dx action, had shown a weak (2-fold) decrease in sensitivity to LE. Circumvention of drug resistance by LE in HL-60/adr cells was accompanied by a 2-fold higher level of H2O2 compared to the wild-type cells, but the mitochondrial respiratory chain inhibitors had no impact on this phenomenon. Conclusions. LE-induced cell death is accompanied by massive hydrogen peroxide production, consisting of two peaks – major one at 1h and secondary at 12h after drug treatment. The highest ROS production was observed in HL-60/adr cells, which have shown a 2-fold increase of resistance to LE compared to parental cells. However, mitochondria seem to play secondary role in this process, due to the fact that the use of specific inhibitors of mitochondrial respiratory chain did not affect the early ROS burst, induced by LE in tumor cells.
Keywords: landomycin E, ROS, mitochondria, cancer drug resistance, apoptosis
Full text: (PDF, in English)

References

[1] Gottesman MM, Fojo T, Bates SE. Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer. 2002;2(1):48-58.
[2] Young RC. Drug resistance: the clinical problem. Cancer Treat Res. 1989;48:1-12.
[3] Ford JM, Hait WN. Pharmacology of drugs that alter multidrug resistance in cancer. Pharmacol Rev. 1990;42(3):155-99.
[4] Jamroziak K, Robak T. Pharmacogenomics of MDR1/ABCB1 gene: the influence on risk and clinical outcome of haematological malignancies. Hematology. 2004;9(2):91-105.
[5] Leighton JC Jr, Goldstein LJ. P-glycoprotein in adult solid tumors. Expression and prognostic significance. Hematol Oncol Clin North Am. 1995;9(2):251-73.
[6] Eberl S, Renner B, Neubert A, Reisig M, Bachmakov I, König J, Dörje F, Mürdter TE, Ackermann A, Dormann H, Gassmann KG, Hahn EG, Zierhut S, Brune K, Fromm MF. Role of p-glycoprotein inhibition for drug interactions: evidence from in vitro and pharmacoepidemiological studies. Clin Pharmacokinet. 2007;46(12):1039-49.
[7] Munoz M, Henderson M, Haber M, Norris M. Role of the MRP1/ABCC1 multidrug transporter protein in cancer. IUBMB Life. 2007;59(12):752-7.
[8] Kodaira H, Kusuhara H, Ushiki J, Fuse E, Sugiyama Y. Kinetic analysis of the cooperation of P-glycoprotein (P-gp/Abcb1) and breast cancer resistance protein (Bcrp/Abcg2) in limiting the brain and testis penetration of erlotinib, flavopiridol, and mitoxantrone. J Pharmacol Exp Ther. 2010;333(3):788-96.
[9] Henkel T, Rohr J, Beale JM, Schwenen L. Landomycins, new angucycline antibiotics from Streptomyces sp. I. Structural studies on landomycins A-D. J Antibiot (Tokyo). 1990;43(5):492-503.
[10] Luzhetskyy A, Zhu L, Gibson M, Fedoryshyn M, Dürr C, Hofmann C, Hoffmeister D, Ostash B, Mattingly C, Adams V, Fedorenko V, Rohr J, Bechthold A. Generation of novel landomycins M and O through targeted gene disruption. Chembiochem. 2005;6(4):675-8.
[11] Shaaban KA, Srinivasan S, Kumar R, Damodaran C, Rohr J. Landomycins P-W, cytotoxic angucyclines from Streptomyces cyanogenus S-136. J Nat Prod. 2011;74(1):2-11.
[12] Panchuk R, Korynevska A, Ostash B, Osyp Y, Fedorenko V, Stoika R. Study of mechanisms of landomycin E action on mammalian cells. Visn L’viv Univ Ser Biol 2004; 35:54-9.
[13] Korynevska A, Heffeter P, Matselyukh B, Elbling L, Micksche M, Stoika R, Berger W. Mechanisms underlying the anticancer activities of the angucycline landomycin E. Biochem Pharmacol. 2007;74(12):1713-26.
[14] Lehka LV, Panchuk RR, Berger W, Rohr J, Stoika RS. The role of reactive oxygen species in tumor cells apoptosis induced by landomycin A. Ukr Biochem J. 2015;87(5):72-82.
[15] Dröse S, Brandt U. Molecular mechanisms of superoxide production by the mitochondrial respiratory chain. Adv Exp Med Biol. 2012;748:145-69.
[16] Riganti C, Gazzano E, Polimeni M, Costamagna C, Bosia A, Ghigo D. Diphenyleneiodonium inhibits the cell redox metabolism and induces oxidative stress. J Biol Chem. 2004;279(46):47726-31.
[17] Chen Q, Vazquez EJ, Moghaddas S, Hoppel CL, Lesnefsky EJ. Production of reactive oxygen species by mitochondria: central role of complex III. J Biol Chem. 2003;278(38):36027-31.
[18] Shchepina LA, Pletjushkina OY, Avetisyan AV, Bakeeva LE, Fetisova EK, Izyumov DS, Saprunova VB, Vyssokikh MY, Chernyak BV, Skulachev VP. Oligomycin, inhibitor of the F0 part of H+-ATP-synthase, suppresses the TNF-induced apoptosis. Oncogene. 2002;21(53):8149-57.
[19] Jagtap JC, Chandele A, Chopde BA, Shastry P. Sodium pyruvate protects against H(2)O(2) mediated apoptosis in human neuroblastoma cell line-SK-N-MC. J Chem Neuroanat. 2003;26(2):109-18.
[20] Samuni Y, Goldstein S, Dean OM, Berk M. The chemistry and biological activities of N-acetylcysteine. Biochim Biophys Acta. 2013;1830(8):4117-29. Review.
[21] Minotti G, Menna P, Salvatorelli E, Cairo G, Gianni L. Anthracyclines: molecular advances and pharmacologic developments in antitumor activity and cardiotoxicity. Pharmacol Rev. 2004;56(2):185-229.
[22] Liou GY, Storz P. Reactive oxygen species in cancer. Free Radic Res. 2010;44(5):479-96.
[23] Lutzky J, Astor MB, Taub RN, Baker MA, Bhalla K, Gervasoni JE Jr, Rosado M, Stewart V, Krishna S, Hindenburg AA. Role of glutathione and dependent enzymes in anthracycline-resistant HL60/AR cells. Cancer Res. 1989;49(15):4120-5.
[24] Legrand O, Zittoun R, Marie JP. Role of MRP1 in multidrug resistance in acute myeloid leukemia. Leukemia. 1999;13(4):578-84.
[25] Versantvoort CH, Broxterman HJ, Bagrij T, Scheper RJ, Twentyman PR. Regulation by glutathione of drug transport in multidrug-resistant human lung tumour cell lines overexpressing multidrug resistance-associated protein. Br J Cancer. 1995;72(1):82-9.