Inhibition of focal adhesion kinase (FAK) activity prevents anchorage-independent ovarian carcinoma cell growth and tumor progression


Recurrence and spread of ovarian cancer is the 5th leading cause of death for women in the United States. Focal adhesion kinase (FAK) is a cytoplasmic protein-tyrosine kinase located on chromosome 8q24.3 (gene is Ptk2), a site commonly amplified in serous ovarian cancer. Elevated FAK mRNA levels in serous ovarian carcinoma are associated with decreased (logrank P = 0.0007, hazard ratio 1.43) patient overall survival, but how FAK functions in tumor progression remains undefined. We have isolated aggressive ovarian carcinoma cells termed ID8-IP after intraperitoneal (IP) growth of murine ID8 cells in C57Bl6 mice. Upon orthotopic implantation within the peri-ovarian bursa space, ID8-IP cells exhibit greater tumor growth, local and distant metastasis, and elevated numbers of ascites-associated cells compared to parental ID8 cells. ID8-IP cells exhibit enhanced growth under non-adherent conditions with elevated FAK and c-Src tyrosine kinase activation compared to parental ID8 cells. In vitro, the small molecule FAK inhibitor (Pfizer, PF562,271, PF-271) at 0.1 uM selectively prevented anchorage-independent ID8-IP cell growth with the inhibition of FAK tyrosine (Y)397 but not c-Src Y416 phosphorylation. Oral PF-271 administration (30 mg/kg, twice daily) blocked FAK but not c-Src tyrosine phosphorylation in ID8-IP tumors. This was associated with decreased tumor size, prevention of peritoneal metastasis, reduced tumor-associated endothelial cell number, and increased tumor cell-associated apoptosis. FAK knockdown and re-expression assays showed that FAK activity selectively promoted anchorage-independent ID8-IP cell survival. These results support the continued evaluation of FAK inhibitors as a promising clinical treatment for ovarian cancer.

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Analysis of variance


Bovine serum albumin




Ethylenediaminetetraacetic acid


Fluorescence-activated cell sorting


Focal adhesion kinase


Fetal bovine serum


Fluorescein isothiocyanate


Green fluorescent protein




Kinase dead


Optimal cutting temperature


Phosphate-buffered saline










Short hairpin RNA


Terminal deoxynucleotidyl transferase dUTP nick end labeling


Wild type


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We appreciate the generosity of Joan Wyllie and “Nine Girls Ask?” for the purchase of a ViCell XR cell viability analyzer and a Leica CM1950 cryostat used in this study. PND-1186 was obtained from Poniard Pharmaceuticals. This work was supported by National Institutes of Health grants (CA102310 and GM087400) to D.D. Schlaepfer. K. Ward is a fellow of the UCSD Reproductive Medicine Gynecological Oncology program. I. Tancioni was supported by a grant from Susan G. Komen for the Cure (KG111237), C. Lawson was supported by a Canadian Institutes of Health Research fellowship (200810MFE-193594-139144), N.L.G. Miller was supported by a National Research Service Award (1F32CA159558), C. Jean was supported by an American Heart Association fellowship (12POST11760014), and D. Stupack was supported by National Institutes of Health grant (CA107263). The authors declare that they have no conflict of interest.

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Corresponding author

Correspondence to David D. Schlaepfer.

Additional information

Kristy K. Ward and Isabelle Tancioni contributed equally to this study.

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Figure S1

Pharmacological FAK inhibition affects growth of human ovarian cancer cell lines. IGROV1, SVOK3, HEY, A2780, and 2008 cells were grown in suspended conditions on ultra-low adherence poly-HEMA plates and treated with 1 μM FAK inhibitor (PND-1186) or DMSO (control) for 72 hours. Values are means (+/- SD) performed in triplicate (**p<0.01; ***p<0.001) (TIF 8840 kb)

Figure S2

Oral administration of PND-1186 FAK inhibitor prevents ID8-IP tumor growth. (A) mCherry-labeled ID8-IP tumor cells were bursal-injected and after 7 days, vehicle or PND-1186 (150 mg/kg) were administered by oral gavage twice-daily (BID). Mice were euthanized after 28 days and (A) primary tumor weight was determined for mice treated with vehicle (n=9) or PND-1186 (n=9). (B) Peritoneal-associated metastatic tumor sites were quantified by counting mCherry-positive nodules as visualized by OV100 imaging. (A and B) Values are means (+/- SD) (* p<0.05, ** p<0.01). (C) Evaluation of FAK Y397 phosphorylation (pY397), total FAK, and total actin levels by immunoblotting using normal ovary tissue or ID8-IP tumors from 5 independent vehicle- or PND-1186-treated mice. (D) Ratio of pY397 phosphorylated FAK to total FAK levels normalized to normal ovary (set to 1) and determined by densitometry using Image J (n= 5 per group, *** p<0.001) (TIF 15656 kb)

Figure S3

Transient GFP expression in Scrambled (Scr) and anti-FAK shRNA ID8-IP cells. Flow cytometry shows GFP (open histogram) or background cell fluorescence (dark histogram) following lentiviral transduction and cell expansion (TIF 15377 kb)

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Ward, K.K., Tancioni, I., Lawson, C. et al. Inhibition of focal adhesion kinase (FAK) activity prevents anchorage-independent ovarian carcinoma cell growth and tumor progression. Clin Exp Metastasis 30, 579–594 (2013).

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  • Ovarian cancer
  • Focal adhesion kinase
  • Cell survival
  • Anchorage-independent cell growth
  • Orthotopic tumor growth
  • Metastasis
  • Syngeneic tumor model