title: The downregulation of Mcl-1 via USP9X inhibition sensitizes solid tumors to Bcl-xl inhibition.
abstract: BACKGROUND
  It has been shown in many solid tumors that the overexpression of the pro-survival Bcl-2 family members Bcl-xL and Mcl-1 confers resistance to a variety of chemotherapeutic agents. Mcl-1 is a critical survival protein in a variety of cell lineages and is critically regulated via ubiquitination.
  
  
  METHODS
  The Mcl-1, Bcl-xL and USP9X expression patterns in human lung and colon adenocarcinomas were evaluated via immunohistochemistry. Interaction between USP9X and Mcl-1 was demonstrated by immunoprecipitation-western blotting. The protein expression profiles of Mcl-1, Bcl-xL and USP9X in multiple cancer cell lines were determined by western blotting. Annexin-V staining and cleaved PARP western blotting were used to assay for apoptosis. The cellular toxicities after various treatments were measured via the XTT assay.
  
  
  RESULTS
  In our current analysis of colon and lung cancer samples, we demonstrate that Mcl-1 and Bcl-xL are overexpressed and also co-exist in many tumors and that the expression levels of both genes correlate with the clinical staging. The downregulation of Mcl-1 or Bcl-xL via RNAi was found to increase the sensitivity of the tumor cells to chemotherapy. Furthermore, our analyses revealed that USP9X expression correlates with that of Mcl-1 in human cancer tissue samples. We additionally found that the USP9X inhibitor WP1130 promotes Mcl-1 degradation and increases tumor cell sensitivity to chemotherapies. Moreover, the combination of WP1130 and ABT-737, a well-documented Bcl-xL inhibitor, demonstrated a chemotherapeutic synergy and promoted apoptosis in different tumor cells.
  
  
  CONCLUSIONS
  Mcl-1, Bcl-xL and USP9X overexpression are tumor survival mechanisms protective against chemotherapy. USP9X inhibition increases tumor cell sensitivity to various chemotherapeutic agents including Bcl-2/Bcl-xL inhibitors.
authors.affiliation: Department of Surgery, Scott & White Memorial Hospital and Clinic, The Texas A&M University System, Health Science Center, College of Medicine, Temple, TX 76504, USA.
authors.forename: Chander
authors.initials: C
authors.lastname: Peddaboina
authors.forename: Daniel
authors.initials: D
authors.lastname: Jupiter
authors.forename: Steven
authors.initials: S
authors.lastname: Fletcher
authors.forename: Jeremy L
authors.initials: JL
authors.lastname: Yap
authors.forename: Arun
authors.initials: A
authors.lastname: Rai
authors.forename: Richard P
authors.initials: RP
authors.lastname: Tobin
authors.forename: Weihua
authors.initials: W
authors.lastname: Jiang
authors.forename: Philip
authors.initials: P
authors.lastname: Rascoe
authors.forename: M Karen Newell
authors.initials: MK
authors.lastname: Rogers
authors.forename: W Roy
authors.initials: WR
authors.lastname: Smythe
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Cao
mesh_terms: D000230:Adenocarcinoma / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D015153:Blotting, Western; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000378:metabolism; D015536:Down-Regulation; D019008:Drug Resistance, Neoplasm / Q000502:physiology; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D008175:Lung Neoplasms / Q000378:metabolism; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis / Q000096:biosynthesis*; D043222:Ubiquitin Thiolesterase / Q000032:analysis / Q000096:biosynthesis*; D051020:bcl-X Protein / Q000032:analysis / Q000096:biosynthesis*
chemical_list: D000970:Antineoplastic Agents; C494811:BCL2L1 protein, human; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D019253:Proto-Oncogene Proteins c-bcl-2; C506234:USP9X protein, human; D051020:bcl-X Protein; D043222:Ubiquitin Thiolesterase