title: JAK-mediated autophagy regulates stemness and cell survival in cisplatin resistant bladder cancer cells.
abstract: Autophagy is a critical process in acquiring drug resistance in solid tumors. However, the mechanisms by which autophagy modulate resistance to chemotherapy in bladder cancer remains poorly understood.
  
  
  
  We have established cisplatin resistant patient derived primary cultured cells as well as T24 bladder cancer cells. The autophagy flux as well as the effect of chemotherapeutic agents, gemcitabine (GC) and mitomycin (MM) were evaluated in these cells. Side population analysis, tumorisphere assay and expression of stemness genes were also studied to evaluate the role of autophagy during chemoresistance. The role of cytokines in autophagy induced chemoresistance was demonstrated.
  
  
  
  Our results show that resistant cells have higher basal autophagic flux. GC and MM led to increase in autophagy and combination of autophagy inhibitors synergistically inhibited bladder cancer cell growth. When autophagy was inhibited, only IFN-γ level was decreased in cytokine array. Blocking IFN-γ decreased JAK2 expression and reduced the autophagy. Inhibition of JAK2 decreased the proportion of cancer stem cell enriched side population, tumor sphere forming ability and led to decrease in expression of stemness genes. Combined treatment with inhibitors of JAK2 and autophagy led to inhibition in cell growth, reduced the levels of inflammatory cytokines and decreased the resistance gene expression.
  
  
  
  GC and MM treatment increased autophagy process via IFN-γ mediated JAK2 and STAT3 pathway.
  
  
  
  These findings support a key role of the autophagy in drug resistance. Thus, autophagy-based, "customized" combinatorial approaches may effectively manage resistance and relapse in bladder cancer.
authors.affiliation: Department of Urology, Post Graduate Institute of Medical Education and Research, Chandigarh 160010032, India.
authors.forename: R
authors.initials: R
authors.lastname: Ojha
authors.affiliation: Department of Urology, Post Graduate Institute of Medical Education and Research, Chandigarh 160010032, India.
authors.forename: S K
authors.initials: SK
authors.lastname: Singh
authors.affiliation: Department of Biophysics, Post Graduate Institute of Medical Education and Research, Chandigarh 160012, India. Electronic address: shalmoli2007@yahoo.co.in.
authors.forename: S
authors.initials: S
authors.lastname: Bhattacharyya
mesh_terms: D000970:Antineoplastic Agents / Q000633:toxicity; D001343:Autophagy; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000633:toxicity; D019008:Drug Resistance, Neoplasm; D006801:Humans; D053614:Janus Kinase 2 / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000378:metabolism; D001749:Urinary Bladder Neoplasms / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; C507924:JAK2 protein, human; D053614:Janus Kinase 2; D002945:Cisplatin
keywords: Autophagy; Bladder cancer; Chemotherapy; Inflammation; JAK; Resistance; Stem cells