title: Hepatic stellate cell coculture enables sorafenib resistance in Huh7 cells through HGF/c-Met/Akt and Jak2/Stat3 pathways.
abstract: OBJECTIVE
  Tumor microenvironment confers drug resistance to kinase inhibitors by increasing RKT ligand levels that result in the activation of cell-survival signaling including PI3K and MAPK signals. We assessed whether HSC-LX2 coculture conferred sorafenib resistance in Huh7 and revealed the mechanism underlying the drug resistance.
  
  
  METHODS
  The effect of LX2 on sorafenib resistance was determined by coculture system with Huh7 cells. The rescue function of LX2 supernatants was assessed by MTT assay and fluorescence microscopy. The underlying mechanism was tested by administration of pathway inhibitors and manifested by Western blotting.
  
  
  RESULTS
  LX2 coculture significantly induced sorafenib resistance in Huh7 by activating p-Akt that led to reactivation of p-ERK. LX2 secreted HGF into the culture medium that triggered drug resistance, and exogenous HGF could also induce sorafenib resistance. The inhibition of p-Akt blocked sorafenib resistance caused by LX2 coculture. Increased phosphorylation of Jak2 and Stat3 was also detected in LX2 cocultured Huh7 cells. The Jak inhibitor tofacitinib reversed sorafenib resistance by blocking Jak2 and Stat3 activation. The combined administration of sorafenib and p-Stat3 inhibitor S3I-201 augmented induced apoptosis even in the presence of sorafenib resistance.
  
  
  CONCLUSIONS
  HSC-LX2 coculture induced sorafenib resistance in Huh7 through multiple pathways: HGF/c-Met/Akt pathway and Jak2/Stat3 pathway. A combined administration of sorafenib and S3I-201 was able to augment sorafenib-induced apoptosis even in the presence of LX2 coculture.
authors: Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital of Nanjing University Medical School, 321 Zhongshan Road, Nanjing, Jiangsu 210008, China ; Jiangsu Province's Key Medical Center for Hepatobiliary Surgery, 321 Zhongshan Road, Nanjing, Jiangsu 210008, China.
authors: Weibo
authors: W
authors: Chen
authors: School of Medicine, Nanjing University, Nanjing, Jiangsu 210093, China.
authors: Junhua
authors: J
authors: Wu
authors: School of Medicine, Nanjing University, Nanjing, Jiangsu 210093, China.
authors: Hua
authors: H
authors: Shi
authors: Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital of Nanjing University Medical School, 321 Zhongshan Road, Nanjing, Jiangsu 210008, China ; Jiangsu Province's Key Medical Center for Hepatobiliary Surgery, 321 Zhongshan Road, Nanjing, Jiangsu 210008, China.
authors: Zhongxia
authors: Z
authors: Wang
authors: Jiangsu Province's Key Medical Center for Hepatobiliary Surgery, 321 Zhongshan Road, Nanjing, Jiangsu 210008, China ; Department of Hepatobiliary Surgery, Drum Tower Clinical College of Nanjing Medical University, Nanjing, Jiangsu 210008, China.
authors: Guang
authors: G
authors: Zhang
authors: Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital of Nanjing University Medical School, 321 Zhongshan Road, Nanjing, Jiangsu 210008, China ; Jiangsu Province's Key Medical Center for Hepatobiliary Surgery, 321 Zhongshan Road, Nanjing, Jiangsu 210008, China.
authors: Yin
authors: Y
authors: Cao
authors: Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital of Nanjing University Medical School, 321 Zhongshan Road, Nanjing, Jiangsu 210008, China ; Jiangsu Province's Key Medical Center for Hepatobiliary Surgery, 321 Zhongshan Road, Nanjing, Jiangsu 210008, China ; Department of Hepatobiliary Surgery, Drum Tower Clinical College of Nanjing Medical University, Nanjing, Jiangsu 210008, China.
authors: Chunping
authors: C
authors: Jiang
authors: Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital of Nanjing University Medical School, 321 Zhongshan Road, Nanjing, Jiangsu 210008, China ; Jiangsu Province's Key Medical Center for Hepatobiliary Surgery, 321 Zhongshan Road, Nanjing, Jiangsu 210008, China ; Department of Hepatobiliary Surgery, Drum Tower Clinical College of Nanjing Medical University, Nanjing, Jiangsu 210008, China.
authors: Yitao
authors: Y
authors: Ding
mesh_terms: D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival; D018920:Coculture Techniques; D003470:Culture Media / Q000737:chemistry; D019008:Drug Resistance, Neoplasm; D004797:Enzyme-Linked Immunosorbent Assay; D055166:Hepatic Stellate Cells / Q000166:cytology*; D006801:Humans; D053614:Janus Kinase 2 / Q000378:metabolism; D008024:Ligands; D008113:Liver Neoplasms / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009536:Niacinamide / Q000031:analogs & derivatives* / Q000737:chemistry; D010671:Phenylurea Compounds / Q000737:chemistry*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D019859:Proto-Oncogene Proteins c-met / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction; D000077157:Sorafenib
chemical_list: D003470:Culture Media; D008024:Ligands; D010671:Phenylurea Compounds; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D009536:Niacinamide; D000077157:Sorafenib; D019859:Proto-Oncogene Proteins c-met; C507924:JAK2 protein, human; D053614:Janus Kinase 2; C494918:AKT1 protein, human; D051057:Proto-Oncogene Proteins c-akt