title: Autophagy in anti-apoptotic effect of augmenter of liver regeneration in HepG2 cells.
abstract: OBJECTIVE
  To investigate the role of autophagy in the anti-apoptotic effect of augmenter of liver regeneration (ALR).
  
  
  METHODS
  Autophagy was induced through serum deprivation. An ALR-expressing plasmid was transfected into HepG2 cells, and autophagic flux was determined using fluorescence microscopy, electron microscopy, Western blot and quantitative polymerase chain reaction (qPCR) assays. After ALR-expressing plasmid transfection, an autophagy inhibitor [3-methyladenine (3-MA)] was added to HepG2 cells, and apoptosis was observed using fluorescence microscopy and flow cytometry.
  
  
  RESULTS
  Autophagy was activated in HepG2 cells, peaking at 24 h after serum deprivation. Microtubule-associated protein light chain three-II levels were higher in HepG2 cells treated with ALR than in control cells, fluorescence microscopy, electron microscopy and qPCR studies showed the similar trend, and p62 levels showed the opposite trend, which indicated that ALR may play an important role in increasing autophagy flux. The numbers of apoptotic cells were substantially higher in HepG2 cells treated with both ALR and 3-MA than in cells treated with ALR alone. Therefore, the protective effect of ALR was significantly attenuated or abolished when autophagy was inhibited, indicating that the anti-apoptotic effect of ALR may be related to autophagy.
  
  
  CONCLUSIONS
  ALR protects cells from apoptosis partly through increased autophagy in HepG2 cells and may be valuable as a new therapeutic treatment for liver disease.
authors.affiliation: Hong-Bo Shi, Hai-Qing Sun, Hong-Lin Shi, Feng Ren, Yu Chen, De-Xi Chen, Jin-Li Lou, Zhong-Ping Duan, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China.
authors.forename: Hong-Bo
authors.initials: HB
authors.lastname: Shi
authors.affiliation: Hong-Bo Shi, Hai-Qing Sun, Hong-Lin Shi, Feng Ren, Yu Chen, De-Xi Chen, Jin-Li Lou, Zhong-Ping Duan, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China.
authors.forename: Hai-Qing
authors.initials: HQ
authors.lastname: Sun
authors.affiliation: Hong-Bo Shi, Hai-Qing Sun, Hong-Lin Shi, Feng Ren, Yu Chen, De-Xi Chen, Jin-Li Lou, Zhong-Ping Duan, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China.
authors.forename: Hong-Lin
authors.initials: HL
authors.lastname: Shi
authors.affiliation: Hong-Bo Shi, Hai-Qing Sun, Hong-Lin Shi, Feng Ren, Yu Chen, De-Xi Chen, Jin-Li Lou, Zhong-Ping Duan, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Ren
authors.affiliation: Hong-Bo Shi, Hai-Qing Sun, Hong-Lin Shi, Feng Ren, Yu Chen, De-Xi Chen, Jin-Li Lou, Zhong-Ping Duan, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Hong-Bo Shi, Hai-Qing Sun, Hong-Lin Shi, Feng Ren, Yu Chen, De-Xi Chen, Jin-Li Lou, Zhong-Ping Duan, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China.
authors.forename: De-Xi
authors.initials: DX
authors.lastname: Chen
authors.affiliation: Hong-Bo Shi, Hai-Qing Sun, Hong-Lin Shi, Feng Ren, Yu Chen, De-Xi Chen, Jin-Li Lou, Zhong-Ping Duan, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China.
authors.forename: Jin-Li
authors.initials: JL
authors.lastname: Lou
authors.affiliation: Hong-Bo Shi, Hai-Qing Sun, Hong-Lin Shi, Feng Ren, Yu Chen, De-Xi Chen, Jin-Li Lou, Zhong-Ping Duan, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China.
authors.forename: Zhong-Ping
authors.initials: ZP
authors.lastname: Duan
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D001343:Autophagy / Q000187:drug effects; D000071187:Autophagy-Related Protein 5; D000071193:Autophagy-Related Protein 7; D000071186:Beclin-1; D003579:Cytochrome Reductases / Q000235:genetics / Q000378:metabolism*; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D050862:Oxidoreductases Acting on Sulfur Group Donors; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D044764:Ubiquitin-Activating Enzymes / Q000235:genetics / Q000378:metabolism
chemical_list: C502778:ATG5 protein, human; D051017:Apoptosis Regulatory Proteins; D000071187:Autophagy-Related Protein 5; C491997:BECN1 protein, human; D000071186:Beclin-1; C477012:MAP1LC3A protein, human; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; C025946:3-methyladenine; D003579:Cytochrome Reductases; C500897:GFER protein, human; D050862:Oxidoreductases Acting on Sulfur Group Donors; C509349:ATG7 protein, human; D000071193:Autophagy-Related Protein 7; D044764:Ubiquitin-Activating Enzymes; D000225:Adenine
keywords: Apoptosis; Augmenter of liver regeneration; Autophagy; HepG2 cells