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Malignant pleural mesothelioma ( MPM ), a highly refractory tumor, is currently incurable due to the lack of an early diagnosis method and medication, both of which are urgently needed to improve the survival and/or quality of life of patients.   NF 2  is a tumor suppressor gene and is frequently mutated in  MPM . Using a  CRISPR /Cas9 system, we generated an   NF 2‐ knockout human mesothelial cell line, MeT‐5A ( NF 2‐ KO ). In  NF 2‐ KO  cell clones, cell growth, clonogenic activity, migration activity, and invasion activity significantly increased compared with those in  NF 2‐ WT  cell clones. Complementary  DNA  microarray analysis clearly revealed the differences in global gene expression profile between  NF 2‐ WT  and  NF 2‐ KO  cell clones. Quantitative  PCR  analysis and western blot analysis showed that the upregulation of fibroblast growth factor receptor 2 ( FGFR 2) was concomitant with the increases in phosphorylation levels of  JNK , c‐Jun, and retinoblastoma (Rb) in  NF 2‐ KO  cell clones. These increases were all abrogated by the exogenous expression of  NF 2 in the  NF 2‐ KO  clone. In addition, the disruption of   FGFR 2  in the  NF 2‐ KO  cell clone suppressed cell proliferation as well as the phosphorylation levels of  JNK , c‐Jun, and Rb. Notably,  FGFR 2 was found to be highly expressed in  NF 2‐negative human mesothelioma tissues (11/12 cases, 91.7%) but less expressed in  NF 2‐positive tissues. Collectively, these findings suggest that   NF 2  deficiency might play a role in the tumorigenesis of human mesothelium through mediating  FGFR 2 expression;  FGFR 2 would be a candidate molecule to develop therapeutic and diagnostic strategies for targeting  MPM  with   NF 2  loss.

cancer science

Establishment and characterization of  CRISPR /Cas9‐mediated   NF 2   −/−   human mesothelial cell line: Molecular insight into fibroblast growth factor receptor 2 in malignant pleural mesothelioma