Prodigiosin induced the caspase-dependent apoptosis in human chronic myelogenous leukemia K562 cell

Maryam Niakani , Ahmad Majd, Parviz Pakzad, Hassan Malekinejad

Abstract


Background and purpose: Chronic myeloid leukemia (CML) as a myeloproliferative disease is characterized by increased cellularity of bone marrow. Implementing the latest treatment protocols is currently accompanied by serious and life-threatening side effects. There are worldwide attempts to find new effective and potent therapeutic agents with minimal side effects on CML patients. This in vitro study was carried out to discover the potential antiproliferative and apoptotic effects of naturally produced prodigiosin (PDG) on K562 cells as an accepted model of CML.

Experimental approach: The anti-proliferative effect of PDG was measured by MTT assay. To highlight the mechanism of cytotoxicity, the apoptotic cell death pathway was investigated by morphological and biochemical assessments. The dual acridine orange/ethidium bromide staining technique and western blotting method were applied to assess the mechanism of the potential apoptotic impact of PDG on K562 cells.

Findings/Results: PDG-induced time- and concentration-dependent anti-proliferative effects were revealed with an estimated IC50 value of 54.06 µM. The highest cell viability reduction (60%) was recorded in cells, which were exposed to 100 µM concentration. Further assays demonstrated that in the dual acridine orange/ethidium bromide staining method the cell population in the late apoptosis phase was increased in a concentration-dependent manner, which was confirmed with remarkable DNA fragmentation.

Conclusion and implications: We found that the PDG-induced apoptosis in K562 cells is mediated through the caspase-3 activation both in mRNA and protein levels. Our results suggest that PDG could be a potent compound for further pharmacokinetic and pharmacodynamics studies in the in vivo model of CML.

 

 


Keywords


Apoptosis; Caspase-3; DNA fragmentation; Prodigiosin.

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References


Koohi F, Salehiniya H, Shamlou R, Eslami S, Ghojogh ZM, et al. Leukemia in Iran: epidemiology and morphology trends. Asian Pac J Cancer Prev. 2015;16(17):7759-7763.

DOI: 10.7314/apjcp.2015.16.17.7759.

Höglund M, Sandin F, Simonsson B. Epidemiology of chronic myeloid leukaemia: an update. Ann Hematol. 2015;94(Suppl 2):S241-S247.

DOI: 10.1007/s00277-015-2314-2.

Saffar A, Rahgozar M, Shahi F, Biglarian A. Survival analysis of acute myeloid leukemia. Razi J Med Sci. 2015;22(134):41-48.

Delluc S, Tourneur L, Michallet AS, Boix C, Varet B, Fradelizi D, et al. Autologous peptides eluted from acute myeloid leukemia cells can be used to generate specific antileukemic CD4 helper and CD8 cytotoxic T lymphocyte responses in vitro. Haematologica. 2005;90(8):1050-1062.

O'Dwyer M. Multifaceted approach to the treatment of bcr-abl-positive leukemias. Oncologist. 2002;7(Suppl 1):30-38.

DOI: 10.1634/theoncologist.7-suppl_1-30.

Deininger MW, Goldman JM, Melo JV. The molecular biology of chronic myeloid leukemia. Blood. 2000;96(10):3343-3356.

Bosch GJ, Joosten AM, Kessler JH, Melief CJ, Leeksma OC. Recognition of BCR-ABL positive leukemic blasts by human CD4+ T cells elicited by primary in vitro immunization with a BCR-ABL breakpoint peptide. Blood. 1996;88(9):3522-3527.

Löffler H, Bergmann J, Hochhaus A, Hehlmann R, Krämer A, et al. Reduced risk for chronic myelogenous leukemia in individuals with the cytochrome P-450 gene polymorphism CYP1A1* 2A. Blood. 2001;98(13):3874-3875.

DOI: 10.1182/blood.V98.13.3874.

Zand AM, Imani S, Saadati MO, Borna H, Ziaei R, Honari HO. Effect of age, gender and blood group on blood cancer types. Kowsar Med J. 2010;15(2):111-114.

Francisco R, Pérez-Tomás R, Gimènez-Bonafé P, Soto-Cerrato V, Giménez-Xavier P, Ambrosio S. Mechanisms of prodigiosin cytotoxicity in human neuroblastoma cell lines. Eur J Pharmacol. 2007;572(2-3):111-119.

DOI: 10.1016/j.ejphar.2007.06.054.

Ho TF, Peng YT, Chuang SM, Lin SC, Feng BL, Lu CH, et al. Prodigiosin down-regulates survivin to facilitate paclitaxel sensitization in human breast carcinoma cell lines. Toxicol Appl Pharmacol. 2009;235(2):253-260.

DOI: 10.1016/j.taap.2008.12.009.

He B, Lu N, Zhou Z. Cellular and nuclear degradation during apoptosis. Curr Opin Cell Biol. 2009;21(6):900-912.

DOI: 10.1016/j.ceb.2009.08.008.

Reed JC. Mechanisms of apoptosis. Am J Pathol. 2000;157(5):1415-1430.

;157(5):1415-1430.

DOI: 10.1016/S0002-9440(10)64779-7.

O’Donovan N, Crown J, Stunell H, Hill ADK, McDermott E, O’Higgins N, et al. Caspase 3 in breast cancer. Clin Cancer Res. 2003;9(2):738-742.

McIlwain DR, Berger T, Mak TW. Caspase functions in cell death and disease. Cold Spring Harb Perspect Biol. 2013;5(4):a008656,1-28.

DOI: 10.1101/cshperspect.a008656.

Samarghandian S, Shabestari MM. DNA fragmentation and apoptosis induced by safranal in human prostate cancer cell line. Indian J Urol. 2013;29(3):177-183.

DOI: 10.4103/0970-1591.117278.

Kalındemirtaş FD, Birman H, Candöken E, Gazioğlu SB, Melikoğlu G, Kuruca S. Cytotoxic effects of some flavonoids and imatinib on the K562 chronic myeloid leukemia cell line: data analysis using the combination index method. Balkan Med J. 2019;36(2):96-105.

DOI: 10.4274/balkanmedj.galenos.2018.2017.1244.

Samejima K, Tone S, Earnshaw WC. CAD/DFF40 nuclease is dispensable for high molecular weight DNA cleavage and stage I chromatin condensation in apoptosis. J Biol Chem. 2001;276(48):45427- 45432.

DOI: 10.1074/jbc.M108844200.

Samejima K, Earnshaw WC. Trashing the genome: the role of nucleases during apoptosis. Nat Rev Mol Cell Biol. 2005;6(9):677-688.

DOI: 10.1038/nrm1715.

Cao G, Pei W, Lan J, Stetler RA, Luo Y, Nagayama T, et al. Caspase-activated DNase/DNA fragmentation factor 40 mediates apoptotic DNA fragmentation in transient cerebral ischemia and in neuronal cultures. J Neurosci. 2001;21(13):4678-4690.

DOI: 10.1523/JNEUROSCI.21-13-04678.2001.

Agarwal A, Mahfouz RZ, Sharma RK, Sarkar O, Mangrola D, Mathur PP. Potential biological role of poly (ADP-ribose) polymerase (PARP) in male gametes. Reprod Biol Endocrinol. 2009;7(1):143-162.

DOI: 10.1186/1477-7827-7-143.

Kamada S, Kikkawa U, Tsujimoto Y, Hunter T. Nuclear translocation of caspase-3 is dependent on its proteolytic activation and recognition of a substrate-like protein (s). J Biol Chem. 2005;280(2):857-860.

DOI: 10.1074/jbc.C400538200.

Pandi SK, Arul D, Smiline AS, Hairul IV, Saravanan SVG, Raghuraman R. Prodigiosin induced apoptosis and inhibited proliferation in carcinoma Hsc-2 cells. Int J Curr Res. 2011;3(4):151-156.

Hassankhani R, Sam MR, Esmaeilou M, Ahangar P. Prodigiosin isolated from cell wall of Serratiamarcescens alters expression of apoptosis-related genes and increases apoptosis in colorectal cancer cells. Med Oncol. 2015;32(1): 366-373.

DOI: 10.1007/s12032-014-0366-0.


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