Combined Treatment of MCF-7 Cells with AICAR and Methotrexate, Arrests Cell Cycle and Reverses Warburg Metabolism through AMP-Activated Protein Kinase (AMPK) and FOXO1.

Fodor T, Szántó M, Abdul-Rahman O, Nagy L, Dér Á, Kiss B, Bai P

PloS one26 Feb 2016Cited by 51 (Europe PMC)

PMID26919657PMCIDPMC4769015DOI10.1371/journal.pone.0150232PepBankPUB-26919657

Summary

Cancer cells are characterized by metabolic alterations, namely, depressed mitochondrial oxidation, enhanced glycolysis and pentose phosphate shunt flux to support rapid cell growth, which is called the Warburg effect. In our study we assessed the metabolic consequences of a joint treatment of MCF-7 breast cancer cells with AICAR, an inducer of AMP-activated kinase (AMPK) jointly with methotrexate (MTX), a folate-analog antimetabolite that blunts de novo nucleotide synthesis. MCF7 cells, a model of breast cancer cells, were resistant to the individual application of AICAR or MTX, however combined treatment of AICAR and MTX reduced cell proliferation. Prolonged joint application of AICAR and MTX induced AMPK and consequently enhanced mitochondrial oxidation and reduced the rate of glycolysis. These metabolic changes suggest an anti-Warburg rearrangement of metabolism that led to the block of the G1/S and the G2/M transition slowing down cell cycle. The slowdown of cell proliferation was abolished when mitotropic transcription factors, PGC-1α, PGC-1β or FOXO1 were silenced. In human breast cancers higher expression of AMPKα and FOXO1 extended survival. AICAR and MTX exerts similar additive antiproliferative effect on other breast cancer cell lines, such as SKBR and 4T1 cells, too. Our data not only underline the importance of Warburg metabolism in breast cancer cells but nominate the AICAR+MTX combination as a potential cytostatic regime blunting Warburg metabolism. Furthermore, we suggest the targeting of AMPK and FOXO1 to combat breast cancer.

CC BYText from Europe PMC. Highlighted sentences are the source of results.

Results from this study12

2 results from studies that gave the peptide. No result from this study comes from people.

01LabIn vitro (cells)Activity measurement
In MCF-7 cells, AICAR alone did not reduce cell proliferation.
In plain wordsIn cells, AICAR alone did not reduce breast cancer cell growth.
Peptide
AICAR (acadesine)
Outcome
No significant changeCell proliferation with AICAR alone
Study
n: not reported in source
MCF-7 breast cancer cells
Outcome group
Tumor growth · Cancer
Studied dose
Dose not stated
Source sentences
Summary
MCF7 cells, a model of breast cancer cells, were resistant to the individual application of AICAR or MTX, however combined treatment of AICAR and MTX reduced cell proliferation.
1 context sentence(design, population, supporting numbers)
Summary · context
In our study we assessed the metabolic consequences of a joint treatment of MCF-7 breast cancer cells with AICAR, an inducer of AMP-activated kinase (AMPK) jointly with methotrexate (MTX), a folate-analog antimetabolite that blunts de novo nucleotide synthesis.
Show in the study text Report a problemFND-2691965701
02LabIn vitro (cells)Activity measurement
In MCF-7 cells, combined AICAR and methotrexate reduced proliferation.
In plain wordsIn cells, AICAR with methotrexate reduced breast cancer cell growth.
Peptide
AICAR (acadesine)
Outcome
DecreaseCell proliferation with combination
Study
n: not reported in source
MCF-7 breast cancer cells
Outcome group
Tumor growth · Cancer
Studied dose
Dose not stated
Source sentences
Summary
MCF7 cells, a model of breast cancer cells, were resistant to the individual application of AICAR or MTX, however combined treatment of AICAR and MTX reduced cell proliferation.
1 context sentence(design, population, supporting numbers)
Summary · context
In our study we assessed the metabolic consequences of a joint treatment of MCF-7 breast cancer cells with AICAR, an inducer of AMP-activated kinase (AMPK) jointly with methotrexate (MTX), a folate-analog antimetabolite that blunts de novo nucleotide synthesis.
Show in the study text Report a problemFND-2691965702

Studies that did not test the peptide alone10

In this study, AICAR (acadesine) was a lab tool or a carrier, part of a mix, or the subject of a lab method. These results are not evidence about taking AICAR (acadesine).

03LabNot a test of AICAR (acadesine) aloneIn vitro (cells)Mechanism
In MCF-7 cells, prolonged combined AICAR and methotrexate induced AMPK.
In plain wordsIn cells, prolonged AICAR with methotrexate induced AMPK.
Peptide
AICAR (acadesine)
Outcome
IncreaseAMPK induction
Study
n: not reported in source
MCF-7 breast cancer cells
Outcome group
Receptor activity and mechanism · Cancer
Source sentences
Summary
Prolonged joint application of AICAR and MTX induced AMPK and consequently enhanced mitochondrial oxidation and reduced the rate of glycolysis.
1 context sentence(design, population, supporting numbers)
Summary · context
In our study we assessed the metabolic consequences of a joint treatment of MCF-7 breast cancer cells with AICAR, an inducer of AMP-activated kinase (AMPK) jointly with methotrexate (MTX), a folate-analog antimetabolite that blunts de novo nucleotide synthesis.
In this study, AICAR (acadesine) was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking AICAR (acadesine).
Show in the study text Report a problemFND-2691965703
04LabNot a test of AICAR (acadesine) aloneIn vitro (cells)Mechanism
In MCF-7 cells, prolonged combined AICAR and methotrexate enhanced mitochondrial oxidation.
In plain wordsIn cells, prolonged AICAR with methotrexate increased mitochondrial oxidation.
Peptide
AICAR (acadesine)
Outcome
IncreaseMitochondrial oxidation
Study
n: not reported in source
MCF-7 breast cancer cells
Outcome group
Mitochondrial function · Cancer
Source sentences
Summary
Prolonged joint application of AICAR and MTX induced AMPK and consequently enhanced mitochondrial oxidation and reduced the rate of glycolysis.
1 context sentence(design, population, supporting numbers)
Summary · context
In our study we assessed the metabolic consequences of a joint treatment of MCF-7 breast cancer cells with AICAR, an inducer of AMP-activated kinase (AMPK) jointly with methotrexate (MTX), a folate-analog antimetabolite that blunts de novo nucleotide synthesis.
In this study, AICAR (acadesine) was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking AICAR (acadesine).
Show in the study text Report a problemFND-2691965704
05LabNot a test of AICAR (acadesine) aloneIn vitro (cells)Mechanism
In MCF-7 cells, prolonged combined AICAR and methotrexate reduced glycolysis rate.
In plain wordsIn cells, prolonged AICAR with methotrexate reduced glycolysis.
Peptide
AICAR (acadesine)
Outcome
DecreaseGlycolysis rate
Study
n: not reported in source
MCF-7 breast cancer cells
Outcome group
Receptor activity and mechanism · Cancer
Source sentences
Summary
Prolonged joint application of AICAR and MTX induced AMPK and consequently enhanced mitochondrial oxidation and reduced the rate of glycolysis.
1 context sentence(design, population, supporting numbers)
Summary · context
In our study we assessed the metabolic consequences of a joint treatment of MCF-7 breast cancer cells with AICAR, an inducer of AMP-activated kinase (AMPK) jointly with methotrexate (MTX), a folate-analog antimetabolite that blunts de novo nucleotide synthesis.
In this study, AICAR (acadesine) was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking AICAR (acadesine).
Show in the study text Report a problemFND-2691965705
06LabNot a test of AICAR (acadesine) aloneIn vitro (cells)Mechanism
In MCF-7 cells, metabolic changes with AICAR and methotrexate were linked to blockade of the G1/S transition.
In plain wordsIn cells, metabolic changes with AICAR and methotrexate were linked to blocked G1/S cell-cycle transition.
Peptide
AICAR (acadesine)
Outcome
DecreaseG1/S transition
Study
n: not reported in source
MCF-7 breast cancer cells
Outcome group
Tumor growth · Cancer
Source sentences
Summary
These metabolic changes suggest an anti-Warburg rearrangement of metabolism that led to the block of the G1/S and the G2/M transition slowing down cell cycle.
1 context sentence(design, population, supporting numbers)
Summary · context
In our study we assessed the metabolic consequences of a joint treatment of MCF-7 breast cancer cells with AICAR, an inducer of AMP-activated kinase (AMPK) jointly with methotrexate (MTX), a folate-analog antimetabolite that blunts de novo nucleotide synthesis.
In this study, AICAR (acadesine) was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking AICAR (acadesine).
Show in the study text Report a problemFND-2691965706
07LabNot a test of AICAR (acadesine) aloneIn vitro (cells)Mechanism
In MCF-7 cells, metabolic changes with AICAR and methotrexate were linked to blockade of the G2/M transition.
In plain wordsIn cells, metabolic changes with AICAR and methotrexate were linked to blocked G2/M cell-cycle transition.
Peptide
AICAR (acadesine)
Outcome
DecreaseG2/M transition
Study
n: not reported in source
MCF-7 breast cancer cells
Outcome group
Tumor growth · Cancer
Source sentences
Summary
These metabolic changes suggest an anti-Warburg rearrangement of metabolism that led to the block of the G1/S and the G2/M transition slowing down cell cycle.
1 context sentence(design, population, supporting numbers)
Summary · context
In our study we assessed the metabolic consequences of a joint treatment of MCF-7 breast cancer cells with AICAR, an inducer of AMP-activated kinase (AMPK) jointly with methotrexate (MTX), a folate-analog antimetabolite that blunts de novo nucleotide synthesis.
In this study, AICAR (acadesine) was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking AICAR (acadesine).
Show in the study text Report a problemFND-2691965707
08LabNot a test of AICAR (acadesine) aloneIn vitro (cells)Mechanism
In MCF-7 cells, PGC-1α silencing abolished the proliferation slowdown with AICAR and methotrexate.
In plain wordsIn cells, PGC-1α silencing blocked the slowing of cell growth with AICAR and methotrexate.
Peptide
AICAR (acadesine)
Outcome
MixedPGC-1α dependence of antiproliferative effect
Study
n: not reported in source
MCF-7 breast cancer cells
Outcome group
Receptor activity and mechanism · Cancer
Source sentences
Summary
The slowdown of cell proliferation was abolished when mitotropic transcription factors, PGC-1α, PGC-1β or FOXO1 were silenced.
1 context sentence(design, population, supporting numbers)
Summary · context
In our study we assessed the metabolic consequences of a joint treatment of MCF-7 breast cancer cells with AICAR, an inducer of AMP-activated kinase (AMPK) jointly with methotrexate (MTX), a folate-analog antimetabolite that blunts de novo nucleotide synthesis.
In this study, AICAR (acadesine) was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking AICAR (acadesine).
Show in the study text Report a problemFND-2691965708
09LabNot a test of AICAR (acadesine) aloneIn vitro (cells)Mechanism
In MCF-7 cells, PGC-1β silencing abolished the proliferation slowdown with AICAR and methotrexate.
In plain wordsIn cells, PGC-1β silencing blocked the slowing of cell growth with AICAR and methotrexate.
Peptide
AICAR (acadesine)
Outcome
MixedPGC-1β dependence of antiproliferative effect
Study
n: not reported in source
MCF-7 breast cancer cells
Outcome group
Receptor activity and mechanism · Cancer
Source sentences
Summary
The slowdown of cell proliferation was abolished when mitotropic transcription factors, PGC-1α, PGC-1β or FOXO1 were silenced.
1 context sentence(design, population, supporting numbers)
Summary · context
In our study we assessed the metabolic consequences of a joint treatment of MCF-7 breast cancer cells with AICAR, an inducer of AMP-activated kinase (AMPK) jointly with methotrexate (MTX), a folate-analog antimetabolite that blunts de novo nucleotide synthesis.
In this study, AICAR (acadesine) was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking AICAR (acadesine).
Show in the study text Report a problemFND-2691965709
10LabNot a test of AICAR (acadesine) aloneIn vitro (cells)Mechanism
In MCF-7 cells, FOXO1 silencing abolished the proliferation slowdown with AICAR and methotrexate.
In plain wordsIn cells, FOXO1 silencing blocked the slowing of cell growth with AICAR and methotrexate.
Peptide
AICAR (acadesine)
Outcome
MixedFOXO1 dependence of antiproliferative effect
Study
n: not reported in source
MCF-7 breast cancer cells
Outcome group
Receptor activity and mechanism · Cancer
Source sentences
Summary
The slowdown of cell proliferation was abolished when mitotropic transcription factors, PGC-1α, PGC-1β or FOXO1 were silenced.
1 context sentence(design, population, supporting numbers)
Summary · context
In our study we assessed the metabolic consequences of a joint treatment of MCF-7 breast cancer cells with AICAR, an inducer of AMP-activated kinase (AMPK) jointly with methotrexate (MTX), a folate-analog antimetabolite that blunts de novo nucleotide synthesis.
In this study, AICAR (acadesine) was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking AICAR (acadesine).
Show in the study text Report a problemFND-2691965710
11LabNot a test of AICAR (acadesine) aloneIn vitro (cells)Activity measurement
In SKBR cells, AICAR and methotrexate had an additive antiproliferative effect.
In plain wordsIn cells, AICAR and methotrexate had an additive effect that reduced SKBR breast cancer cell growth.
Peptide
AICAR (acadesine)
Outcome
DecreaseSKBR cell proliferation
Study
n: not reported in source
SKBR breast cancer cells
Outcome group
Tumor growth · Cancer
Source sentences
Summary
AICAR and MTX exerts similar additive antiproliferative effect on other breast cancer cell lines, such as SKBR and 4T1 cells, too.
In this study, AICAR (acadesine) was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking AICAR (acadesine).
Show in the study text Report a problemFND-2691965711
12LabNot a test of AICAR (acadesine) aloneIn vitro (cells)Activity measurement
In 4T1 cells, AICAR and methotrexate had an additive antiproliferative effect.
In plain wordsIn cells, AICAR and methotrexate had an additive effect that reduced 4T1 breast cancer cell growth.
Peptide
AICAR (acadesine)
Outcome
Decrease4T1 cell proliferation
Study
n: not reported in source
4T1 breast cancer cells
Outcome group
Tumor growth · Cancer
Source sentences
Summary
AICAR and MTX exerts similar additive antiproliferative effect on other breast cancer cell lines, such as SKBR and 4T1 cells, too.
In this study, AICAR (acadesine) was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking AICAR (acadesine).
Show in the study text Report a problemFND-2691965712

AI wrote and checked these results. AI can make mistakes, so check important results against the source.

Version history

PepBank keeps every version of the data on this study. Status changes from Retraction Watch, Europe PMC and Crossref are listed with their source.

Version 2Current
21 Oct 2026, 15:58 UTC
Study results updated
Version 1
21 Oct 2026, 12:50 UTC
First added · source update from Europe PMC

PepBank is a research reference. Talk to a doctor before you use any peptide.

Combined Treatment of MCF-7 Cells with AICAR and Methotrexate, Arrests Cell Cycle and Reverses Warburg Metabolism through AMP-Activated Protein Kinase (AMPK) and FOXO1.

Summary

Results from this study12

Studies that did not test the peptide alone10

Version history

Subjects

Publication types

MeSH terms