TY  - JOUR
AU  - Misirkić Marjanović, Maja
AU  - Janjetović, Kristina
AU  - Vučićević, Ljubica
AU  - Tovilović-Kovačević, Gordana
AU  - Ristić, Biljana Z
AU  - Vilimanović, Uros
AU  - Harhaji-Trajković, Ljubica
AU  - Sumarac-Dumanović, Mirjana S
AU  - Micić, Dragan D
AU  - Bumbaširević, Vladimir Z
AU  - Trajković, Vladimir S
PY  - 2012
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1232
AB  - The role of autophagy, a process in which the cell self-digests its own components, was investigated in glioma cell death induced by the hydroxymethylglutaryl-coenzyme A (HMG-CoA) reductase-inhibiting drug simvastatin. Induction of autophagy and activation of autophagy-regulating signalling pathways were analyzed by immunoblotting. Flow cytometry/fluorescent microscopy was used to assess autophagy-associated intracellular acidification and apoptotic markers (phosphatidylserine exposure, DNA fragmentation and caspase activation). Cell viability was determined by crystal violet, MTT or LDH release assay. Simvastatin treatment of U251 and C6 glioma cell lines caused the appearance of autophagolysosome-like intracytoplasmic acidic vesicles. The induction of autophagy in U251 cells was confirmed by the upregulation of autophagosome-associated LC3-II and pro-autophagic beclin-1, as well as by the downregulation of the selective autophagic target p62. Simvastatin induced the activation of AMP-activated protein kinase (AMPK) and its target Raptor, while simultaneously downregulating activation of Akt. Mammalian target of rapamycin (mTOR). a major AMPK/Akt downstream target and a major negative autophagy regulator, and its substrate p70 S6 kinase 1 were also inhibited by simvastatin. Mevalonate, the product of HMG-CoA reductase enzymatic activity, AMPK siRNA or pharmacological inactivation of AMPK with compound C suppressed, while the inhibitors of Akt (10-DEBC hydrochloride) and mTOR (rapamycin) mimicked autophagy induction by simvastatin. Inhibition of autophagy with bafilomycin A1, 3-methyladenine and LC3 beta shRNA, as well as AMPK inhibition with compound C or AMPK siRNA, markedly increased apoptotic death of simvastatin-treated U251 cells. These data suggest that inhibition of AMPK-depenclent autophagic response might sensitize glioma cells to statin-induced apoptotic death. (C) 2011 Elsevier Ltd. All rights reserved.
T2  - Pharmacological Research
T1  - Inhibition of AMPK-dependent autophagy enhances in vitro antiglioma effect of simvastatin
IS  - 1
VL  - 65
EP  - 119
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1232
ER  - 

@article{
author = "Misirkić Marjanović, Maja and Janjetović, Kristina and Vučićević, Ljubica and Tovilović-Kovačević, Gordana and Ristić, Biljana Z and Vilimanović, Uros and Harhaji-Trajković, Ljubica and Sumarac-Dumanović, Mirjana S and Micić, Dragan D and Bumbaširević, Vladimir Z and Trajković, Vladimir S",
year = "2012",
abstract = "The role of autophagy, a process in which the cell self-digests its own components, was investigated in glioma cell death induced by the hydroxymethylglutaryl-coenzyme A (HMG-CoA) reductase-inhibiting drug simvastatin. Induction of autophagy and activation of autophagy-regulating signalling pathways were analyzed by immunoblotting. Flow cytometry/fluorescent microscopy was used to assess autophagy-associated intracellular acidification and apoptotic markers (phosphatidylserine exposure, DNA fragmentation and caspase activation). Cell viability was determined by crystal violet, MTT or LDH release assay. Simvastatin treatment of U251 and C6 glioma cell lines caused the appearance of autophagolysosome-like intracytoplasmic acidic vesicles. The induction of autophagy in U251 cells was confirmed by the upregulation of autophagosome-associated LC3-II and pro-autophagic beclin-1, as well as by the downregulation of the selective autophagic target p62. Simvastatin induced the activation of AMP-activated protein kinase (AMPK) and its target Raptor, while simultaneously downregulating activation of Akt. Mammalian target of rapamycin (mTOR). a major AMPK/Akt downstream target and a major negative autophagy regulator, and its substrate p70 S6 kinase 1 were also inhibited by simvastatin. Mevalonate, the product of HMG-CoA reductase enzymatic activity, AMPK siRNA or pharmacological inactivation of AMPK with compound C suppressed, while the inhibitors of Akt (10-DEBC hydrochloride) and mTOR (rapamycin) mimicked autophagy induction by simvastatin. Inhibition of autophagy with bafilomycin A1, 3-methyladenine and LC3 beta shRNA, as well as AMPK inhibition with compound C or AMPK siRNA, markedly increased apoptotic death of simvastatin-treated U251 cells. These data suggest that inhibition of AMPK-depenclent autophagic response might sensitize glioma cells to statin-induced apoptotic death. (C) 2011 Elsevier Ltd. All rights reserved.",
journal = "Pharmacological Research",
title = "Inhibition of AMPK-dependent autophagy enhances in vitro antiglioma effect of simvastatin",
number = "1",
volume = "65",
pages = "119",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1232"
}

Misirkić Marjanović, M., Janjetović, K., Vučićević, L., Tovilović-Kovačević, G., Ristić, B. Z., Vilimanović, U., Harhaji-Trajković, L., Sumarac-Dumanović, M. S., Micić, D. D., Bumbaširević, V. Z.,& Trajković, V. S.. (2012). Inhibition of AMPK-dependent autophagy enhances in vitro antiglioma effect of simvastatin. in Pharmacological Research, 65(1).
https://hdl.handle.net/21.15107/rcub_ibiss_1232

Misirkić Marjanović M, Janjetović K, Vučićević L, Tovilović-Kovačević G, Ristić BZ, Vilimanović U, Harhaji-Trajković L, Sumarac-Dumanović MS, Micić DD, Bumbaširević VZ, Trajković VS. Inhibition of AMPK-dependent autophagy enhances in vitro antiglioma effect of simvastatin. in Pharmacological Research. 2012;65(1):null-119.
https://hdl.handle.net/21.15107/rcub_ibiss_1232 .

Misirkić Marjanović, Maja, Janjetović, Kristina, Vučićević, Ljubica, Tovilović-Kovačević, Gordana, Ristić, Biljana Z, Vilimanović, Uros, Harhaji-Trajković, Ljubica, Sumarac-Dumanović, Mirjana S, Micić, Dragan D, Bumbaširević, Vladimir Z, Trajković, Vladimir S, "Inhibition of AMPK-dependent autophagy enhances in vitro antiglioma effect of simvastatin" in Pharmacological Research, 65, no. 1 (2012),
https://hdl.handle.net/21.15107/rcub_ibiss_1232 .

TY  - JOUR
AU  - Misirlić-Dencić, Sonja T
AU  - Poljarević, Jelena M
AU  - Vilimanović, Uros
AU  - Bogdanović, Andrija D
AU  - Isaković, Aleksandra J
AU  - Kravić-Stevović, Tamara K
AU  - Dulović, Marija
AU  - Zogović, Nevena
AU  - Isaković, Anđelka M
AU  - Grgurić-Sipka, Sanja R
AU  - Bumbaširević, Vladimir Z
AU  - Sabo, Tibor J
AU  - Trajković, Vladimir S
AU  - Marković, Ivanka D
PY  - 2012
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1196
AB  - We investigated the cytotoxicity of recently synthesized (S,S)-ethyleridiamine-N,N'-di-2-(3-cyclohexyl)propanoic acid esters toward human leukemic cell lines and healthy blood mononuclear cells. Cell viability was assessed by acid phosphatase assay, apoptosis, and differentiation were analyzed by flow cytometry and electron microscopy, while intracellular localization of apoptosis-inducing factor (AIF) was determined by immunoblotting. It was demonstrated that methyl, ethyl, and n-propyl esters were toxic to HL-60, REH, MOLT-4, KG-1, JVM-2, and K-562 leukemic cell lines, while the nonesterified parental compound and n-butyl ester were devoid of cytotoxic action. The ethyl ester exhibited the highest cytotoxic activity (IC50 10.7 mu M-45.4 mu M), which was comparable to that of the prototypical anticancer drug cisplatin. The observed cytotoxic effect in HL-60 cells was associated with an increase in superoxide production and mitochondrial membrane depolarization, leading to apoptotic cell death characterized by phosphatidylserine externalization and DNA fragmentation in the absence of autophagic response. DNA fragmentation preceded caspase activation and followed AIF translocation from mitochondria to nucleus, which was indicative of caspase-independent apoptotic cell death. HL-60 cells treated with subtoxic concentration of the compound displayed morphological signs of granulocytic differentiation (nuclear indentations and presence of cytoplasmic primary granules), as well as an increased expression of differentiation markers CD11b and CD15. The cyclohexyl analogues of ethylenediamine dipropanoic acid were also toxic to peripheral blood mononuclear cells of both healthy controls and leukemic patients, the latter being more sensitive. Our data demonstrate that the toxicity of the investigated cyclohexyl compounds against leukemic cell lines is mediated by caspase-independent apoptosis associated with oxidative stress, mitochondrial dysfunction, and AIF translocation.
T2  - Chemical Research in Toxicology
T1  - Cyclohexyl Analogues of Ethylenediamine Dipropanoic Acid Induce Caspase-Independent Mitochondrial Apoptosis in Human Leukemic Cells
IS  - 4
VL  - 25
EP  - 939
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1196
ER  - 

@article{
author = "Misirlić-Dencić, Sonja T and Poljarević, Jelena M and Vilimanović, Uros and Bogdanović, Andrija D and Isaković, Aleksandra J and Kravić-Stevović, Tamara K and Dulović, Marija and Zogović, Nevena and Isaković, Anđelka M and Grgurić-Sipka, Sanja R and Bumbaširević, Vladimir Z and Sabo, Tibor J and Trajković, Vladimir S and Marković, Ivanka D",
year = "2012",
abstract = "We investigated the cytotoxicity of recently synthesized (S,S)-ethyleridiamine-N,N'-di-2-(3-cyclohexyl)propanoic acid esters toward human leukemic cell lines and healthy blood mononuclear cells. Cell viability was assessed by acid phosphatase assay, apoptosis, and differentiation were analyzed by flow cytometry and electron microscopy, while intracellular localization of apoptosis-inducing factor (AIF) was determined by immunoblotting. It was demonstrated that methyl, ethyl, and n-propyl esters were toxic to HL-60, REH, MOLT-4, KG-1, JVM-2, and K-562 leukemic cell lines, while the nonesterified parental compound and n-butyl ester were devoid of cytotoxic action. The ethyl ester exhibited the highest cytotoxic activity (IC50 10.7 mu M-45.4 mu M), which was comparable to that of the prototypical anticancer drug cisplatin. The observed cytotoxic effect in HL-60 cells was associated with an increase in superoxide production and mitochondrial membrane depolarization, leading to apoptotic cell death characterized by phosphatidylserine externalization and DNA fragmentation in the absence of autophagic response. DNA fragmentation preceded caspase activation and followed AIF translocation from mitochondria to nucleus, which was indicative of caspase-independent apoptotic cell death. HL-60 cells treated with subtoxic concentration of the compound displayed morphological signs of granulocytic differentiation (nuclear indentations and presence of cytoplasmic primary granules), as well as an increased expression of differentiation markers CD11b and CD15. The cyclohexyl analogues of ethylenediamine dipropanoic acid were also toxic to peripheral blood mononuclear cells of both healthy controls and leukemic patients, the latter being more sensitive. Our data demonstrate that the toxicity of the investigated cyclohexyl compounds against leukemic cell lines is mediated by caspase-independent apoptosis associated with oxidative stress, mitochondrial dysfunction, and AIF translocation.",
journal = "Chemical Research in Toxicology",
title = "Cyclohexyl Analogues of Ethylenediamine Dipropanoic Acid Induce Caspase-Independent Mitochondrial Apoptosis in Human Leukemic Cells",
number = "4",
volume = "25",
pages = "939",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1196"
}

Misirlić-Dencić, S. T., Poljarević, J. M., Vilimanović, U., Bogdanović, A. D., Isaković, A. J., Kravić-Stevović, T. K., Dulović, M., Zogović, N., Isaković, A. M., Grgurić-Sipka, S. R., Bumbaširević, V. Z., Sabo, T. J., Trajković, V. S.,& Marković, I. D.. (2012). Cyclohexyl Analogues of Ethylenediamine Dipropanoic Acid Induce Caspase-Independent Mitochondrial Apoptosis in Human Leukemic Cells. in Chemical Research in Toxicology, 25(4).
https://hdl.handle.net/21.15107/rcub_ibiss_1196

Misirlić-Dencić ST, Poljarević JM, Vilimanović U, Bogdanović AD, Isaković AJ, Kravić-Stevović TK, Dulović M, Zogović N, Isaković AM, Grgurić-Sipka SR, Bumbaširević VZ, Sabo TJ, Trajković VS, Marković ID. Cyclohexyl Analogues of Ethylenediamine Dipropanoic Acid Induce Caspase-Independent Mitochondrial Apoptosis in Human Leukemic Cells. in Chemical Research in Toxicology. 2012;25(4):null-939.
https://hdl.handle.net/21.15107/rcub_ibiss_1196 .

Misirlić-Dencić, Sonja T, Poljarević, Jelena M, Vilimanović, Uros, Bogdanović, Andrija D, Isaković, Aleksandra J, Kravić-Stevović, Tamara K, Dulović, Marija, Zogović, Nevena, Isaković, Anđelka M, Grgurić-Sipka, Sanja R, Bumbaširević, Vladimir Z, Sabo, Tibor J, Trajković, Vladimir S, Marković, Ivanka D, "Cyclohexyl Analogues of Ethylenediamine Dipropanoic Acid Induce Caspase-Independent Mitochondrial Apoptosis in Human Leukemic Cells" in Chemical Research in Toxicology, 25, no. 4 (2012),
https://hdl.handle.net/21.15107/rcub_ibiss_1196 .

TY  - JOUR
AU  - Stevanović, Darko M
AU  - Starčević, Vesna P.
AU  - Vilimanović, Uros
AU  - Nesić, Dejan M
AU  - Vučićević, Ljubica
AU  - Misirkić Marjanović, Maja
AU  - Janjetović, Kristina
AU  - Savić, Emina
AU  - Popadić, Dusan M
AU  - Sudar, Emina M
AU  - Micić, Dragan D
AU  - Sumarac-Dumanović, Mirjana S
AU  - Trajković, Vladimir S
PY  - 2012
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1238
AB  - We investigated the effects of centrally administered orexigenic hormone ghrelin on energy imbalance-induced inflammation. Rats were subjected for four weeks to three different dietary regimes: normal (standard food), high-fat (standard food with 30% lard) or food-restricted (70%, 50%, 40% and 40% of the expected food intake in 1st, 2nd, 3rd and 4th week, respectively). Compared to normal-weight controls, starved, but not obese rats had significantly higher levels of proinflammatory cytokines (TNF, IL-1 beta, IFN-gamma) in the blood. When compared to normally fed animals, the hearts of starved and obese animals expressed higher levels of mRNAs encoding proinflammatory mediators (TNF, IL-1 beta, IL-6, IFN-gamma, IL-17, IL-12, iNOS), while mRNA levels of the anti-inflammatory TGF-beta remained unchanged. Intracerebroventricular (ICV) injection of ghrelin (1 mu g/day) for five consecutive days significantly reduced TNF, IL-1 beta and IFN-gamma levels in the blood of starved rats, as well as TNF, IL-17 and IL-12p40 mRNA expression in the hearts of obese rats. Conversely, ICV ghrelin increased the levels of 1FN-gamma, IL-17,1L-12p35 and IL-12p40 mRNA in the heart tissue of food-restricted animals. This was associated with an increase of immunosuppressive ACTH/corticosterone production in starved animals and a decrease of the immunostimulatory adipokine leptin both in food-restricted and high-fat groups. Ghrelin activated the energy sensor AMP-activated protein kinase (AMPK) in the hypothalamus and inhibited extracellular signal-regulated kinase (ERK) in the hearts of obese, but not starved rats. Therefore, central ghrelin may play a complex role in energy imbalance-induced inflammation by modulating HPA axis, leptin and AMPK/ERK signaling pathways. (C) 2011 Elsevier Inc. All rights reserved.
T2  - Brain Behavior and Immunity
T1  - Immunomodulatory actions of central ghrelin in diet-induced energy imbalance
IS  - 1
VL  - 26
EP  - 158
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1238
ER  - 

@article{
author = "Stevanović, Darko M and Starčević, Vesna P. and Vilimanović, Uros and Nesić, Dejan M and Vučićević, Ljubica and Misirkić Marjanović, Maja and Janjetović, Kristina and Savić, Emina and Popadić, Dusan M and Sudar, Emina M and Micić, Dragan D and Sumarac-Dumanović, Mirjana S and Trajković, Vladimir S",
year = "2012",
abstract = "We investigated the effects of centrally administered orexigenic hormone ghrelin on energy imbalance-induced inflammation. Rats were subjected for four weeks to three different dietary regimes: normal (standard food), high-fat (standard food with 30% lard) or food-restricted (70%, 50%, 40% and 40% of the expected food intake in 1st, 2nd, 3rd and 4th week, respectively). Compared to normal-weight controls, starved, but not obese rats had significantly higher levels of proinflammatory cytokines (TNF, IL-1 beta, IFN-gamma) in the blood. When compared to normally fed animals, the hearts of starved and obese animals expressed higher levels of mRNAs encoding proinflammatory mediators (TNF, IL-1 beta, IL-6, IFN-gamma, IL-17, IL-12, iNOS), while mRNA levels of the anti-inflammatory TGF-beta remained unchanged. Intracerebroventricular (ICV) injection of ghrelin (1 mu g/day) for five consecutive days significantly reduced TNF, IL-1 beta and IFN-gamma levels in the blood of starved rats, as well as TNF, IL-17 and IL-12p40 mRNA expression in the hearts of obese rats. Conversely, ICV ghrelin increased the levels of 1FN-gamma, IL-17,1L-12p35 and IL-12p40 mRNA in the heart tissue of food-restricted animals. This was associated with an increase of immunosuppressive ACTH/corticosterone production in starved animals and a decrease of the immunostimulatory adipokine leptin both in food-restricted and high-fat groups. Ghrelin activated the energy sensor AMP-activated protein kinase (AMPK) in the hypothalamus and inhibited extracellular signal-regulated kinase (ERK) in the hearts of obese, but not starved rats. Therefore, central ghrelin may play a complex role in energy imbalance-induced inflammation by modulating HPA axis, leptin and AMPK/ERK signaling pathways. (C) 2011 Elsevier Inc. All rights reserved.",
journal = "Brain Behavior and Immunity",
title = "Immunomodulatory actions of central ghrelin in diet-induced energy imbalance",
number = "1",
volume = "26",
pages = "158",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1238"
}

Stevanović, D. M., Starčević, V. P., Vilimanović, U., Nesić, D. M., Vučićević, L., Misirkić Marjanović, M., Janjetović, K., Savić, E., Popadić, D. M., Sudar, E. M., Micić, D. D., Sumarac-Dumanović, M. S.,& Trajković, V. S.. (2012). Immunomodulatory actions of central ghrelin in diet-induced energy imbalance. in Brain Behavior and Immunity, 26(1).
https://hdl.handle.net/21.15107/rcub_ibiss_1238

Stevanović DM, Starčević VP, Vilimanović U, Nesić DM, Vučićević L, Misirkić Marjanović M, Janjetović K, Savić E, Popadić DM, Sudar EM, Micić DD, Sumarac-Dumanović MS, Trajković VS. Immunomodulatory actions of central ghrelin in diet-induced energy imbalance. in Brain Behavior and Immunity. 2012;26(1):null-158.
https://hdl.handle.net/21.15107/rcub_ibiss_1238 .

Stevanović, Darko M, Starčević, Vesna P., Vilimanović, Uros, Nesić, Dejan M, Vučićević, Ljubica, Misirkić Marjanović, Maja, Janjetović, Kristina, Savić, Emina, Popadić, Dusan M, Sudar, Emina M, Micić, Dragan D, Sumarac-Dumanović, Mirjana S, Trajković, Vladimir S, "Immunomodulatory actions of central ghrelin in diet-induced energy imbalance" in Brain Behavior and Immunity, 26, no. 1 (2012),
https://hdl.handle.net/21.15107/rcub_ibiss_1238 .

TY  - JOUR
AU  - Vučićević, Ljubica
AU  - Misirkić Marjanović, Maja
AU  - Janjetović, Kristina
AU  - Vilimanović, Uros
AU  - Sudar, Emina M
AU  - Isenović, Esma R
AU  - Prica, Marko
AU  - Harhaji-Trajković, Ljubica
AU  - Kravić-Stevović, Tamara K
AU  - Bumbaširević, Vladimir Z
AU  - Trajković, Vladimir S
PY  - 2011
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1321
AB  - In the present study, we report that compound C, an inhibitor of a key intracellular energy sensor AMP-activated protein kinase (AMPK), can induce autophagy in cancer cells. The induction of autophagy in U251 human glioma cell line was demonstrated by acridine orange staining of intracellular acidic vesicles, Beclin 1 induction, p62 decrease and conversion of LC3-I to autophagosome-associated LC3-II in the presence of proteolysis inhibitors. The presence of autophagosome-like vesicles was confirmed by transmission electron microscopy. Compound C-mediated inhibition of AMPK and raptor in U251 cells was associated with paradoxical decrease in phosphorylation of AMPK/raptor-repressed mTOR, a major negative regulator of autophagy, and its downstream target p70S6K. The phosphorylation of an mTOR activator Akt and the PI3K-activating kinase Src was also impaired in compound C-treated cells. The siRNA-mediated AMPK silencing did not reduce the activity of the Akt/mTOR/p70S6K pathway and AMPK activators metformin and AICAR failed to block compound C-induced autophagy. Autophagy inhibitors bafilomycin and chloroquine significantly increased the cytotoxicity of compound C towards U251 cells, as confirmed by increase in lactate dehydrogenase release, DNA fragmentation and caspase-3 activation. Similar effects of compound C were also observed in C6 rat glioma, L929 mouse fibrosarcoma and B16 mouse melanoma cell lines. Since compound C has previously been reported to suppress AMPK-dependent autophagy in different cell types, our findings suggest that the effects of compound C on autophagy might be dose-, cell type- and/or context-dependent. By demonstrating the ability of compound C to induce autophagic response in cancer cells via AMPK inhibition-independent downregulation of Akt/mTOR pathway, our results warrant caution when using compound C to inhibit AMPK-dependent cellular responses, but also support further exploration of compound C and related molecules as potential anticancer agents.
T2  - Autophagy
T1  - Compound C induces protective autophagy in cancer cells through AMPK inhibition-independent blockade of Akt/mTOR pathway
IS  - 1
VL  - 7
DO  - 10.4161/auto.7.1.13883
EP  - 50
ER  - 

@article{
author = "Vučićević, Ljubica and Misirkić Marjanović, Maja and Janjetović, Kristina and Vilimanović, Uros and Sudar, Emina M and Isenović, Esma R and Prica, Marko and Harhaji-Trajković, Ljubica and Kravić-Stevović, Tamara K and Bumbaširević, Vladimir Z and Trajković, Vladimir S",
year = "2011",
abstract = "In the present study, we report that compound C, an inhibitor of a key intracellular energy sensor AMP-activated protein kinase (AMPK), can induce autophagy in cancer cells. The induction of autophagy in U251 human glioma cell line was demonstrated by acridine orange staining of intracellular acidic vesicles, Beclin 1 induction, p62 decrease and conversion of LC3-I to autophagosome-associated LC3-II in the presence of proteolysis inhibitors. The presence of autophagosome-like vesicles was confirmed by transmission electron microscopy. Compound C-mediated inhibition of AMPK and raptor in U251 cells was associated with paradoxical decrease in phosphorylation of AMPK/raptor-repressed mTOR, a major negative regulator of autophagy, and its downstream target p70S6K. The phosphorylation of an mTOR activator Akt and the PI3K-activating kinase Src was also impaired in compound C-treated cells. The siRNA-mediated AMPK silencing did not reduce the activity of the Akt/mTOR/p70S6K pathway and AMPK activators metformin and AICAR failed to block compound C-induced autophagy. Autophagy inhibitors bafilomycin and chloroquine significantly increased the cytotoxicity of compound C towards U251 cells, as confirmed by increase in lactate dehydrogenase release, DNA fragmentation and caspase-3 activation. Similar effects of compound C were also observed in C6 rat glioma, L929 mouse fibrosarcoma and B16 mouse melanoma cell lines. Since compound C has previously been reported to suppress AMPK-dependent autophagy in different cell types, our findings suggest that the effects of compound C on autophagy might be dose-, cell type- and/or context-dependent. By demonstrating the ability of compound C to induce autophagic response in cancer cells via AMPK inhibition-independent downregulation of Akt/mTOR pathway, our results warrant caution when using compound C to inhibit AMPK-dependent cellular responses, but also support further exploration of compound C and related molecules as potential anticancer agents.",
journal = "Autophagy",
title = "Compound C induces protective autophagy in cancer cells through AMPK inhibition-independent blockade of Akt/mTOR pathway",
number = "1",
volume = "7",
doi = "10.4161/auto.7.1.13883",
pages = "50"
}

Vučićević, L., Misirkić Marjanović, M., Janjetović, K., Vilimanović, U., Sudar, E. M., Isenović, E. R., Prica, M., Harhaji-Trajković, L., Kravić-Stevović, T. K., Bumbaširević, V. Z.,& Trajković, V. S.. (2011). Compound C induces protective autophagy in cancer cells through AMPK inhibition-independent blockade of Akt/mTOR pathway. in Autophagy, 7(1).
https://doi.org/10.4161/auto.7.1.13883

Vučićević L, Misirkić Marjanović M, Janjetović K, Vilimanović U, Sudar EM, Isenović ER, Prica M, Harhaji-Trajković L, Kravić-Stevović TK, Bumbaširević VZ, Trajković VS. Compound C induces protective autophagy in cancer cells through AMPK inhibition-independent blockade of Akt/mTOR pathway. in Autophagy. 2011;7(1):null-50.
doi:10.4161/auto.7.1.13883 .

Vučićević, Ljubica, Misirkić Marjanović, Maja, Janjetović, Kristina, Vilimanović, Uros, Sudar, Emina M, Isenović, Esma R, Prica, Marko, Harhaji-Trajković, Ljubica, Kravić-Stevović, Tamara K, Bumbaširević, Vladimir Z, Trajković, Vladimir S, "Compound C induces protective autophagy in cancer cells through AMPK inhibition-independent blockade of Akt/mTOR pathway" in Autophagy, 7, no. 1 (2011),
https://doi.org/10.4161/auto.7.1.13883 . .