title: Targeting autophagy augments in vitro and in vivo antimyeloma activity of DNA-damaging chemotherapy.
abstract: OBJECTIVE
  Although autophagy occurs in most tumor cells following DNA damage, it is still a mystery how this DNA-damaging event turns on the autophagy machinery in multiple myeloma (MM) and how the functional status of autophagy impacts on its susceptibility to death in response to DNA-damaging chemotherapy.
  
  
  METHODS
  We investigate the effects of DNA damage on autophagy in MM cells and elucidate its underlying molecular mechanism. Then, we examined the impacts of pharmacologic or genetic inhibition of autophagy on DNA damage-induced apoptosis. Furthermore, the antimyeloma activity of autophagy inhibitor in combination with DNA-damaging agents was evaluated in MM xenograft models.
  
  
  RESULTS
  We showed that DNA-damaging drugs, doxorubicin and melphalan, induce caspase-dependent apoptosis and concurrently trigger Beclin 1-regulated autophagy in human MM cell lines H929 and RPMI 8226. Mechanistically, association of autophagy execution proteins Beclin 1 with class III phosphoinositide 3-kinase, which is inhibited by Bcl-2 recruitment, contributes directly to the autophagic process. Importantly, targeting suppression of autophagy by minimally toxic concentrations of pharmacologic inhibitors (hydroxychloroquine and 3-methyladenine) or short hairpin RNAs against autophagy genes, Beclin 1 and Atg5, dramatically augments proapoptotic activity of DNA-damaging chemotherapy both in vitro using MM cell lines or purified patient MM cells and in vivo in a human plasmacytoma xenograft mouse model.
  
  
  CONCLUSIONS
  These data can help unravel the underlying molecular mechanism of autophagy in DNA-damaged MM cells and also provide a rationale for clinical evaluation of autophagy inhibitors in combination with DNA-damaging chemotherapy in MM.
authors.affiliation: Department of Hematology, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, PR China.
authors.forename: Yaozhu
authors.initials: Y
authors.lastname: Pan
authors.forename: Ying
authors.initials: Y
authors.lastname: Gao
authors.forename: Liang
authors.initials: L
authors.lastname: Chen
authors.forename: Guangxun
authors.initials: G
authors.lastname: Gao
authors.forename: Hongjuan
authors.initials: H
authors.lastname: Dong
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.forename: Baoxia
authors.initials: B
authors.lastname: Dong
authors.forename: Xiequn
authors.initials: X
authors.lastname: Chen
mesh_terms: D000225:Adenine / Q000008:administration & dosage / Q000031:analogs & derivatives; D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology / Q000627:therapeutic use*; D001343:Autophagy / Q000187:drug effects*; D045744:Cell Line, Tumor; D004249:DNA Damage / Q000187:drug effects*; D004317:Doxorubicin / Q000008:administration & dosage; D004357:Drug Synergism; D006801:Humans; D008558:Melphalan / Q000008:administration & dosage; D051379:Mice; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D058990:Molecular Targeted Therapy; D009101:Multiple Myeloma / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D023041:Xenograft Model Antitumor Assays
chemical_list: C025946:3-methyladenine; D004317:Doxorubicin; D000225:Adenine; D008558:Melphalan