Blockade of reactive oxygen species and Akt activation is critical for anti-inflammation and growth inhibition of metformin in phosphatase and tensin homolog-deficient RAW264.7 cells

Chiou Feng Lin, Kung Chia Young, Chyi Huey Bai, Bu Chin Yu, Ching Ting Ma, Yu Chieh Chien, Hui Chen Su, Hue Yu Wang, Chao Sheng Liao, Hsin Wen Lai, Chiung Wen Tsao

Research output: Contribution to journalArticle

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Abstract

Context: Metformin is widely used for treatment of type 2 diabetes and has a potential application on the treatment of inflammation and cancer. Phosphatase and tensin homolog (PTEN) plays a critical role in cancer cell growth and inflammation; however, precise mechanisms remain unclear. Objective: We aimed to investigate the possible mechanisms of how PTEN regulates metformin against cell growth and inflammation. Materials and methods: We established PTEN knockdown in RAW264.7 murine macrophages (shPTEN cells) to detect inflammatory mediators using commercial kits, production of reactive oxygen species (ROS) by flow cytometry, cell growth by MTT assay and phosphorylated levels of signal molecules by western blot. Results: The shPTEN cells had a significant large amount of inflammatory mediators, such as inducible nitric oxide synthase (iNOS)/nitric oxide (NO) and cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE2); and also elevated the production of ROS and increased cell proliferation. These effects were accompanied with the activation of Akt and p38 mitogen-activated protein kinase (MAPK), and the inactivation of an AMP-activated protein kinase (AMPK) activator and extracellular signal-regulated kinase 1/2. Pretreatment with metformin not only blocked these inflammatory mediators, but also caused growth inhibition induced by significant apoptosis. Furthermore, inactivation of Akt, blockade of ROS generation and independence of activations of AMPK and MAPK by metformin were also observed. Conclusion: Macrophages with PTEN deficiency developed a continuous inflammatory microenvironment, which further aggravated tumor cell growth. Moreover, metformin affected PTEN-deficient cells dependent of inhibition of ROS production and Akt activation against enlarged inflammatory mediators and/or cell growth in shPTEN cells.

Original languageEnglish
Pages (from-to)669-677
Number of pages9
JournalImmunopharmacology and Immunotoxicology
Volume35
Issue number6
DOIs
Publication statusPublished - Dec 2013
Externally publishedYes

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Metformin
Cell growth
Phosphoric Monoester Hydrolases
Reactive Oxygen Species
Chemical activation
Inflammation
Growth
AMP-Activated Protein Kinases
Macrophages
Mitogen-Activated Protein Kinase 3
Flow cytometry
Cell proliferation
p38 Mitogen-Activated Protein Kinases
Nitric Oxide Synthase Type II
Cyclooxygenase 2
Medical problems
Mitogen-Activated Protein Kinases
Dinoprostone
Tumors
Assays

Keywords

  • COX-2/PGE production
  • INOS/NO release
  • Metformin
  • Phosphatase and tensin homolog (PTEN)
  • Reactive oxygen species

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy
  • Pharmacology
  • Toxicology

Cite this

Blockade of reactive oxygen species and Akt activation is critical for anti-inflammation and growth inhibition of metformin in phosphatase and tensin homolog-deficient RAW264.7 cells. / Lin, Chiou Feng; Young, Kung Chia; Bai, Chyi Huey; Yu, Bu Chin; Ma, Ching Ting; Chien, Yu Chieh; Su, Hui Chen; Wang, Hue Yu; Liao, Chao Sheng; Lai, Hsin Wen; Tsao, Chiung Wen.

In: Immunopharmacology and Immunotoxicology, Vol. 35, No. 6, 12.2013, p. 669-677.

Research output: Contribution to journalArticle

Lin, Chiou Feng ; Young, Kung Chia ; Bai, Chyi Huey ; Yu, Bu Chin ; Ma, Ching Ting ; Chien, Yu Chieh ; Su, Hui Chen ; Wang, Hue Yu ; Liao, Chao Sheng ; Lai, Hsin Wen ; Tsao, Chiung Wen. / Blockade of reactive oxygen species and Akt activation is critical for anti-inflammation and growth inhibition of metformin in phosphatase and tensin homolog-deficient RAW264.7 cells. In: Immunopharmacology and Immunotoxicology. 2013 ; Vol. 35, No. 6. pp. 669-677.
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abstract = "Context: Metformin is widely used for treatment of type 2 diabetes and has a potential application on the treatment of inflammation and cancer. Phosphatase and tensin homolog (PTEN) plays a critical role in cancer cell growth and inflammation; however, precise mechanisms remain unclear. Objective: We aimed to investigate the possible mechanisms of how PTEN regulates metformin against cell growth and inflammation. Materials and methods: We established PTEN knockdown in RAW264.7 murine macrophages (shPTEN cells) to detect inflammatory mediators using commercial kits, production of reactive oxygen species (ROS) by flow cytometry, cell growth by MTT assay and phosphorylated levels of signal molecules by western blot. Results: The shPTEN cells had a significant large amount of inflammatory mediators, such as inducible nitric oxide synthase (iNOS)/nitric oxide (NO) and cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE2); and also elevated the production of ROS and increased cell proliferation. These effects were accompanied with the activation of Akt and p38 mitogen-activated protein kinase (MAPK), and the inactivation of an AMP-activated protein kinase (AMPK) activator and extracellular signal-regulated kinase 1/2. Pretreatment with metformin not only blocked these inflammatory mediators, but also caused growth inhibition induced by significant apoptosis. Furthermore, inactivation of Akt, blockade of ROS generation and independence of activations of AMPK and MAPK by metformin were also observed. Conclusion: Macrophages with PTEN deficiency developed a continuous inflammatory microenvironment, which further aggravated tumor cell growth. Moreover, metformin affected PTEN-deficient cells dependent of inhibition of ROS production and Akt activation against enlarged inflammatory mediators and/or cell growth in shPTEN cells.",
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author = "Lin, {Chiou Feng} and Young, {Kung Chia} and Bai, {Chyi Huey} and Yu, {Bu Chin} and Ma, {Ching Ting} and Chien, {Yu Chieh} and Su, {Hui Chen} and Wang, {Hue Yu} and Liao, {Chao Sheng} and Lai, {Hsin Wen} and Tsao, {Chiung Wen}",
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T1 - Blockade of reactive oxygen species and Akt activation is critical for anti-inflammation and growth inhibition of metformin in phosphatase and tensin homolog-deficient RAW264.7 cells

AU - Lin, Chiou Feng

AU - Young, Kung Chia

AU - Bai, Chyi Huey

AU - Yu, Bu Chin

AU - Ma, Ching Ting

AU - Chien, Yu Chieh

AU - Su, Hui Chen

AU - Wang, Hue Yu

AU - Liao, Chao Sheng

AU - Lai, Hsin Wen

AU - Tsao, Chiung Wen

PY - 2013/12

Y1 - 2013/12

N2 - Context: Metformin is widely used for treatment of type 2 diabetes and has a potential application on the treatment of inflammation and cancer. Phosphatase and tensin homolog (PTEN) plays a critical role in cancer cell growth and inflammation; however, precise mechanisms remain unclear. Objective: We aimed to investigate the possible mechanisms of how PTEN regulates metformin against cell growth and inflammation. Materials and methods: We established PTEN knockdown in RAW264.7 murine macrophages (shPTEN cells) to detect inflammatory mediators using commercial kits, production of reactive oxygen species (ROS) by flow cytometry, cell growth by MTT assay and phosphorylated levels of signal molecules by western blot. Results: The shPTEN cells had a significant large amount of inflammatory mediators, such as inducible nitric oxide synthase (iNOS)/nitric oxide (NO) and cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE2); and also elevated the production of ROS and increased cell proliferation. These effects were accompanied with the activation of Akt and p38 mitogen-activated protein kinase (MAPK), and the inactivation of an AMP-activated protein kinase (AMPK) activator and extracellular signal-regulated kinase 1/2. Pretreatment with metformin not only blocked these inflammatory mediators, but also caused growth inhibition induced by significant apoptosis. Furthermore, inactivation of Akt, blockade of ROS generation and independence of activations of AMPK and MAPK by metformin were also observed. Conclusion: Macrophages with PTEN deficiency developed a continuous inflammatory microenvironment, which further aggravated tumor cell growth. Moreover, metformin affected PTEN-deficient cells dependent of inhibition of ROS production and Akt activation against enlarged inflammatory mediators and/or cell growth in shPTEN cells.

AB - Context: Metformin is widely used for treatment of type 2 diabetes and has a potential application on the treatment of inflammation and cancer. Phosphatase and tensin homolog (PTEN) plays a critical role in cancer cell growth and inflammation; however, precise mechanisms remain unclear. Objective: We aimed to investigate the possible mechanisms of how PTEN regulates metformin against cell growth and inflammation. Materials and methods: We established PTEN knockdown in RAW264.7 murine macrophages (shPTEN cells) to detect inflammatory mediators using commercial kits, production of reactive oxygen species (ROS) by flow cytometry, cell growth by MTT assay and phosphorylated levels of signal molecules by western blot. Results: The shPTEN cells had a significant large amount of inflammatory mediators, such as inducible nitric oxide synthase (iNOS)/nitric oxide (NO) and cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE2); and also elevated the production of ROS and increased cell proliferation. These effects were accompanied with the activation of Akt and p38 mitogen-activated protein kinase (MAPK), and the inactivation of an AMP-activated protein kinase (AMPK) activator and extracellular signal-regulated kinase 1/2. Pretreatment with metformin not only blocked these inflammatory mediators, but also caused growth inhibition induced by significant apoptosis. Furthermore, inactivation of Akt, blockade of ROS generation and independence of activations of AMPK and MAPK by metformin were also observed. Conclusion: Macrophages with PTEN deficiency developed a continuous inflammatory microenvironment, which further aggravated tumor cell growth. Moreover, metformin affected PTEN-deficient cells dependent of inhibition of ROS production and Akt activation against enlarged inflammatory mediators and/or cell growth in shPTEN cells.

KW - COX-2/PGE production

KW - INOS/NO release

KW - Metformin

KW - Phosphatase and tensin homolog (PTEN)

KW - Reactive oxygen species

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