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Cytotoxic activity of the novel heterocyclic compound G-11 is primarily mediated through intrinsic apoptotic pathway

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Abstract

Natural and chemically synthesized heterocyclic compounds have been explored for their potential use as anticancer agents. We had synthesized non-natural heterocyclic analogs and evaluated their anti-tumor activity by measuring effect on cell proliferation and induction of apoptosis in different cell lines. Previously, we identified a pyrazole-containing compound (G-11) showing cytotoxic effect towards leukemia and lymphoma cell lines. In this study, we further investigated the mechanistic aspects of anticancer properties of G-11 in HL-60 cell line. We demonstrated that cytotoxic effect of G-11 is mediated by caspase-dependent apoptosis. However, the involvement of mitochondrial dysfunction induced by G-11 was independent of caspases. G-11 triggered generation of ROS, caused disruption of mitochondrial transmembrane potential, increased release of cytochrome c to the cytosol, and altered the expression of Bcl-2 and Bax proteins. These results suggest significant involvement of intrinsic apoptotic pathway. This study comprehensively details the possible mechanisms of action of a novel heterocyclic compound which could find its potential use as an anticancer agent.

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Abbreviations

G-11:

[1-(2′′,3′′,4′′,6′′-Tetra-O-acetyl-β-D-glucopyranosyl)-4-(3′-trifluoromethylphenyl hydrazono)-3-trifluoromethyl-1,4-dihydropyrazol-5-one]

HL-60:

Human acute myelogenous leukemia cell line

∆ψm :

Difference in mitochondrial transmembrane potential

ROS:

Reactive oxygen species

VDAC:

Voltage dependent anion channel

Bcl-2:

B-cell lymphoma 2

Bcl-xL:

B-cell lymphoma-extra-large

Bax:

Bcl-2-associated X protein

Bak:

Bcl-2 homologous antagonist killer

PARP:

Poly (ADP-ribose) polymerase

7-AAD:

7-amino-actinomycin D

PDTC:

Pyrrolidine dithiocarbamate

NAC:

N-Acetyl cysteine

z-VAD-FMK:

N-Benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone

Necrostatin-1:

[5-(1H-Indol-3-ylmethyl)-3-methyl-2-thioxo-4-Imidazolidinone, 5-(Indol-3-ylmethyl)-3-methyl-2-thio-Hydantoin, MTH-DL-Tryptophan]

DMSO:

Dimethyl sulfoxide

PMSF:

Phenylmethanesulfonyl fluoride

Ac-LEHD-pNA:

N-acetyl-Leu-Glu-His-Asp-p-nitroanilide

Ac-IETD-pNA:

N-acetyl-Ile-Glu-Thr-Asp-p-nitroanilide

Ac-DEVD-pNA:

N-acetyl-Asp-Glu-Val-Asp-p-nitroanilide

PVDF:

Polyvinylidene fluoride

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Acknowledgments

This project is supported by a research Grant from King Abdulaziz City for Science and Technology (KACST; Grant Number- AT-34-136, Riyadh, Kingdom of Saudi Arabia) for Saleh A.M. Publication cost is supported by King Abdullah International Medical Research Center (KAIMRC) in Riyadh, Saudi Arabia.

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Authors and Affiliations

  1. Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Saud Bin Abdulaziz University for Health Sciences, King Abdulaziz Medical City, National Guard Health Affairs, Mail Code 6610, P. O. Box 9515, Jeddah, 21423, Kingdom of Saudi Arabia

    Ayman M. Saleh

  2. King Abdullah International Medical Research Center (KAIMRC), P. O. Box 9515, Jeddah, 21423, Kingdom of Saudi Arabia

    Ayman M. Saleh & Ahmad Aljada

  3. King Abdullah International Medical Research Center (KAIMRC), National Guard Health Affairs, P.O. Box 22490, Riyadh, 11426, Kingdom of Saudi Arabia

    Mohammad A. Aziz

  4. Department of Chemistry, Faculty of Sciences, United Arab Emirates University, P. O. Box 15551, Al-Ain, United Arab Emirates

    Ibrahim M. Abdou

  5. Department of Pharmaceutical Sciences, Faculty of Pharmacy, The University of Jordan, Amman, 11942, Jordan

    Mutasem O. Taha

  6. Department of Chemistry, Faculty of Sciences, Yarmouk University, Irbid, 21163, Jordan

    Mahmoud A. Al-Qudah

  7. Department of Pharmacognosy and Pharmaceutical Chemistry, College of Pharmacy, Taibah University, Al Madinah Almonawarrah, 41477, Saudi Arabia

    Mohammed M. Abadleh

  8. Department of Basic Medical Sciences, College of Medicine, King Saud Bin Abdulaziz University for Health Sciences, Riyadh, 11481, Kingdom of Saudi Arabia

    Ahmad Aljada

  9. Department of Pharmaceutical Sciences, College of Pharmacy, Nova Southeastern University (NSU), Fort Lauderdale, FL, 33328, USA

    Syed A. Rizvi

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  1. Ayman M. Saleh
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  2. Mohammad A. Aziz
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  3. Ibrahim M. Abdou
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  4. Mutasem O. Taha
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  5. Mahmoud A. Al-Qudah
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  6. Mohammed M. Abadleh
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  7. Ahmad Aljada
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  8. Syed A. Rizvi
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Corresponding author

Correspondence to Ayman M. Saleh.

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Saleh, A.M., Aziz, M.A., Abdou, I.M. et al. Cytotoxic activity of the novel heterocyclic compound G-11 is primarily mediated through intrinsic apoptotic pathway. Apoptosis 21, 873–886 (2016). https://doi.org/10.1007/s10495-016-1248-z

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