Blockage of glutamine-dependent anaplerosis affects mTORC1/2 activity and ultimately leads to cellular senescence-like response
The purpose of study was to explore the role of glutamine-dependent anaplerosis in cell fate determination (proliferation and senescence) and the potential associated mechanism by employing a pharmacological inhibitor of glutamine-dependent anaplerosis, amino-oxyacetate (AOA). Using the WI38 normal...
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| Format: | Article |
| Language: | English |
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The Company of Biologists
2019-05-01
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| Series: | Biology Open |
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| Online Access: | http://bio.biologists.org/content/8/5/bio038257 |
| _version_ | 1818671192755666944 |
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| author | Geng-You Liao Ming-Ting Lee Jhen-Jia Fan Pei-Wen Hsiao Chun-Sheng Lee Shou-Yi Su Jiuan-Jiuan Hwang Ferng-Chun Ke |
| author_facet | Geng-You Liao Ming-Ting Lee Jhen-Jia Fan Pei-Wen Hsiao Chun-Sheng Lee Shou-Yi Su Jiuan-Jiuan Hwang Ferng-Chun Ke |
| author_sort | Geng-You Liao |
| collection | DOAJ |
| description | The purpose of study was to explore the role of glutamine-dependent anaplerosis in cell fate determination (proliferation and senescence) and the potential associated mechanism by employing a pharmacological inhibitor of glutamine-dependent anaplerosis, amino-oxyacetate (AOA). Using the WI38 normal human embryonic fibroblast cell line, we found that exposure to AOA induced mTORC1 inactivation−mTORC2 activation (within day 1), cell cycle arrest (day 2–6) and cellular senescence (day 4–6). These AOA effects were blocked by concomitantly providing anaplerotic factors [α-ketoglutarate (αKG), pyruvate or oxaloacetate], and not affected by ROS scavenger N-acetyl-cysteine (NAC). Moreover, AOA-induced cellular senescence in WI38 cells is associated with elevated protein levels of p53, p21CIP1 and p16INK4A and decreased Rb protein level, which was blocked by αKG supplementation. In p16INK4A-deficient U2OS human osteosarcoma cells and p16INK4A-knockdown WI38 cells, AOA exposure also induced similar effects on cell proliferation, and protein level of P-Rb-S807/811 and Rb. Interestingly, no AOA induction of cellular senescence was observed in U2OS cells, yet was still seen in p16INK4A-knockdown WI38 cells accompanied by the presence of p16 antibody-reactive p12. In summary, we disclose that glutamine-dependent anaplerosis is essential to cell growth and closely associated with mTORC1 activation and mTORC2 inactivation, and impedes cellular senescence particularly associated with p16INK4A. |
| first_indexed | 2024-12-17T07:20:06Z |
| format | Article |
| id | doaj.art-628a6b2fe1b74292a21f5aba8c269cdb |
| institution | Directory Open Access Journal |
| issn | 2046-6390 |
| language | English |
| last_indexed | 2024-12-17T07:20:06Z |
| publishDate | 2019-05-01 |
| publisher | The Company of Biologists |
| record_format | Article |
| series | Biology Open |
| spelling | doaj.art-628a6b2fe1b74292a21f5aba8c269cdb2022-12-21T21:58:47ZengThe Company of BiologistsBiology Open2046-63902019-05-018510.1242/bio.038257038257Blockage of glutamine-dependent anaplerosis affects mTORC1/2 activity and ultimately leads to cellular senescence-like responseGeng-You Liao0Ming-Ting Lee1Jhen-Jia Fan2Pei-Wen Hsiao3Chun-Sheng Lee4Shou-Yi Su5Jiuan-Jiuan Hwang6Ferng-Chun Ke7 Institute of Molecular and Cellular Biology, College of Life Science, National Taiwan University, Taipei 106, Taiwan Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan Institute of Molecular and Cellular Biology, College of Life Science, National Taiwan University, Taipei 106, Taiwan Agricultural Biotechnology Research Center, Academia Sinica, Taipei 115, Taiwan Institute of Molecular and Cellular Biology, College of Life Science, National Taiwan University, Taipei 106, Taiwan Institute of Molecular and Cellular Biology, College of Life Science, National Taiwan University, Taipei 106, Taiwan Institute of Physiology, School of Medicine, National Yang-Ming University, Taipei 112, Taiwan Institute of Molecular and Cellular Biology, College of Life Science, National Taiwan University, Taipei 106, Taiwan The purpose of study was to explore the role of glutamine-dependent anaplerosis in cell fate determination (proliferation and senescence) and the potential associated mechanism by employing a pharmacological inhibitor of glutamine-dependent anaplerosis, amino-oxyacetate (AOA). Using the WI38 normal human embryonic fibroblast cell line, we found that exposure to AOA induced mTORC1 inactivation−mTORC2 activation (within day 1), cell cycle arrest (day 2–6) and cellular senescence (day 4–6). These AOA effects were blocked by concomitantly providing anaplerotic factors [α-ketoglutarate (αKG), pyruvate or oxaloacetate], and not affected by ROS scavenger N-acetyl-cysteine (NAC). Moreover, AOA-induced cellular senescence in WI38 cells is associated with elevated protein levels of p53, p21CIP1 and p16INK4A and decreased Rb protein level, which was blocked by αKG supplementation. In p16INK4A-deficient U2OS human osteosarcoma cells and p16INK4A-knockdown WI38 cells, AOA exposure also induced similar effects on cell proliferation, and protein level of P-Rb-S807/811 and Rb. Interestingly, no AOA induction of cellular senescence was observed in U2OS cells, yet was still seen in p16INK4A-knockdown WI38 cells accompanied by the presence of p16 antibody-reactive p12. In summary, we disclose that glutamine-dependent anaplerosis is essential to cell growth and closely associated with mTORC1 activation and mTORC2 inactivation, and impedes cellular senescence particularly associated with p16INK4A.http://bio.biologists.org/content/8/5/bio038257Amino-oxyacetateCellular senescenceGlutamine-dependent anaplerosisMetabolismmTORCp16INK4A |
| spellingShingle | Geng-You Liao Ming-Ting Lee Jhen-Jia Fan Pei-Wen Hsiao Chun-Sheng Lee Shou-Yi Su Jiuan-Jiuan Hwang Ferng-Chun Ke Blockage of glutamine-dependent anaplerosis affects mTORC1/2 activity and ultimately leads to cellular senescence-like response Biology Open Amino-oxyacetate Cellular senescence Glutamine-dependent anaplerosis Metabolism mTORC p16INK4A |
| title | Blockage of glutamine-dependent anaplerosis affects mTORC1/2 activity and ultimately leads to cellular senescence-like response |
| title_full | Blockage of glutamine-dependent anaplerosis affects mTORC1/2 activity and ultimately leads to cellular senescence-like response |
| title_fullStr | Blockage of glutamine-dependent anaplerosis affects mTORC1/2 activity and ultimately leads to cellular senescence-like response |
| title_full_unstemmed | Blockage of glutamine-dependent anaplerosis affects mTORC1/2 activity and ultimately leads to cellular senescence-like response |
| title_short | Blockage of glutamine-dependent anaplerosis affects mTORC1/2 activity and ultimately leads to cellular senescence-like response |
| title_sort | blockage of glutamine dependent anaplerosis affects mtorc1 2 activity and ultimately leads to cellular senescence like response |
| topic | Amino-oxyacetate Cellular senescence Glutamine-dependent anaplerosis Metabolism mTORC p16INK4A |
| url | http://bio.biologists.org/content/8/5/bio038257 |
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