Analyses of merlin/NF2 connection to FAK inhibitor responsiveness in serous ovarian cancer

Gynecol Oncol. 2014 Jul;134(1):104-11. doi: 10.1016/j.ygyno.2014.04.044. Epub 2014 Apr 27.

Abstract

Objective: Focal adhesion kinase (FAK) is overexpressed in serous ovarian cancer. Loss of merlin, a product of the neurofibromatosis 2 tumor suppressor gene, is being evaluated as a biomarker for FAK inhibitor sensitivity in mesothelioma. Connections between merlin and FAK in ovarian cancer remain undefined.

Methods: Nine human and two murine ovarian cancer cell lines were analyzed for growth in the presence of a small molecule FAK inhibitor (PF-271, also termed VS-6062) from 0.1 to 1 μM for 72 h. Merlin was evaluated by immunoblotting and immunostaining of a human ovarian tumor tissue array. Growth of cells was analyzed in an orthotopic tumor model and evaluated in vitro after stable shRNA-mediated merlin knockdown.

Results: Greater than 50% inhibition of OVCAR8, HEY, and ID8-IP ovarian carcinoma cell growth occurred with 0.1 μM PF-271 in anchorage-independent (p<0.001) but not in adherent culture conditions. PF-271-mediated reduction in FAK Y397 phosphorylation occurred independently of growth inhibition. Suspended growth of OVCAR3, OVCAR10, IGROV1, IGROV1-IP, SKOV3, SKOV3-IP, A2780, and 5009-MOVCAR was not affected by 0.1 μM PF-271. Merlin expression did not correlate with serous ovarian tumor grade or stage. PF-271 (30 mg/kg, BID) did not inhibit 5009-MOVCAR tumor growth and merlin knockdown in SKOV3-IP and OVCAR10 cells did not alter suspended cell growth upon PF-271 addition.

Conclusions: Differential responsiveness to FAK inhibitor treatment was observed. Intrinsic low merlin protein level correlated with PF-271-mediated anchorage-independent growth inhibition, but reduction in merlin expression did not induce sensitivity to FAK inhibition. Merlin levels may be useful for patient stratification in FAK inhibitor trials.

Keywords: Biomarker; Focal adhesion kinase; Merlin; Ovarian cancer; Tumor.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers, Tumor / metabolism
  • Cell Line, Tumor
  • Cystadenocarcinoma, Serous / drug therapy*
  • Cystadenocarcinoma, Serous / enzymology
  • Cystadenocarcinoma, Serous / metabolism
  • Female
  • Focal Adhesion Kinase 1 / antagonists & inhibitors*
  • Focal Adhesion Kinase 1 / metabolism
  • Gene Knockdown Techniques
  • Humans
  • Mice
  • Neurofibromin 2 / genetics
  • Neurofibromin 2 / metabolism*
  • Ovarian Neoplasms / drug therapy*
  • Ovarian Neoplasms / enzymology
  • Ovarian Neoplasms / metabolism
  • Protein Kinase Inhibitors / pharmacology*

Substances

  • Biomarkers, Tumor
  • Neurofibromin 2
  • Protein Kinase Inhibitors
  • Focal Adhesion Kinase 1
  • PTK2 protein, human
  • Ptk2 protein, mouse