FGFR2 Is Amplified in the NCI-H716 Colorectal Cancer Cell Line and Is Required for Growth and Survival

Abstract

Aberrant kinase activation resulting from mutation, amplification, or translocation can drive growth and survival in a subset of human cancer. FGFR2 is amplified in breast and gastric cancer, and we report here the first characterization of FGFR2 gene amplification in colorectal cancer in the NCI-H716 colorectal cancer cell line. FGFR2 is highly expressed and activated in NCI-H716 cells, and FGFR selective small molecule inhibitors or FGFR2 shRNA strongly inhibited cell viability in vitro, indicating “addiction” of NCI-H716 cells to FGFR2. NCI-H716 growth in a xenograft model was also inhibited by an FGFR small molecule inhibitor. FGFR2 was required for activation of multiple downstream signaling proteins including AKT, ERK, S6RP and NFKB. Inhibition of downstream kinases such as AKT or ERK alone had modest effects on proliferation, whereas combined inhibition of AKT and ERK signaling resulted in a loss of viability similar to FGFR2 inhibition. We identified elevated FGFR2 expression in a small subset of primary colorectal cancer, however FGFR2 amplification was not observed. Although FGFR2 amplification is not common in primary colon cancer or lymph node and liver metastases, other subsets of colorectal cancer such as ascites, from which the NCI-H716 cell line was derived, have yet to be tested. These results suggest that emerging FGFR inhibitor therapeutics may have efficacy in a subset of colon cancer driven by FGFR2 amplification.

The molecular landscape of colorectal cancer cell lines unveils clinically actionable kinase targets

“…In some instances (FGFR2, ALK or NTRK1), overexpression of the TK gene was associated with amplification or translocation of the corresponding locus. The same genetic alterations have been previously reported in CRC cells and patients [26][27][28] , thus validating our approach. Our data further suggest that overexpression of TK sustains primary resistance to EGFR blockade, and could be used to identify patients unlikely to respond to cetuximab or panitumumab.…”

“…We decided to focus on TKs for which targeted agents are in clinical trial or are already approved for treatment, namely ALK, FGFR2, KIT, NTRK1, NTRK2, RET and PDGFRA. This analysis revealed that the overexpression of NTRK1 and FGFR2 is associated to molecular alterations, such as gene translocation (NTRK1) or gene amplification (FGFR2; Supplementary Figs 5 and 6), previously described in cellular models and in cancer patients [26][27][28] , thus validating the experimental approach. The other TK outlier genes were not previously reported in CRC cells.…”

AZD-4547 exerts potent cytostatic and cytotoxic activities against fibroblast growth factor receptor (FGFR)-expressing colorectal cancer cells

“…FGFR-1 and FGFR-2 were over-expressed in NCI-H716 cells [15], but were undetected in HT-29 cells (Fig. 1a).…”

Exploring the mechanism of WWOX growth inhibitory effects on oral squamous cell carcinoma

“…Katoh and Nakagama (27) demonstrated that the expression of FGFR2 was amplified in breast and gastric cancer. In the colorectal cancer NCI-H716 cell line, which exhibits a high expression of FGFR2, the inhibition of FGFR2 by small molecule inhibitors or FGFR2 short hairpin (sh)RNA was shown to decrease cell viability (28). In pancreatic cancer, tumor cells with FGFR2-shRNA transfection exhibited attenuated proliferation rates, migration and invasion levels, and a reduced level of phosphorylation of ERK compared with that of the control cells (29).…”