Cancer Res Treat.  2020 Oct;52(4):1162-1177. 10.4143/crt.2020.138.

FGFR4 Gly388Arg Polymorphism Affects the Progression of Gastric Cancer by Activating STAT3 Pathway to Induce Epithelial to Mesenchymal Transition

• Ye Y
• Li J
• Jiang D
• Li J
• Xiao C
• Li Y
• Han C
• Zhao C

• KMID: 2507942
• DOI: http://doi.org/10.4143/crt.2020.138

Abstract

Purpose
Fibroblast growth factor receptor 4 (FGFR4) plays a critical role in cancer progression involving in tumor proliferation, invasion, and metastasis. This study clarified the role of FGFR4-Arg388 variant in gastric cancer (GC), and more importantly highlighted the possibility of this single nucleotide polymorphism (SNP) as potential therapeutic targets.
Materials and Methods
FGFR4 polymorphism was characterized in advanced GC patients to perform statistical analysis. FGFR4-dependent signal pathways involving cell proliferation, invasion, migration, and resistance to oxaliplatin (OXA) in accordance with the SNP were also assessed in transfected GC cell lines.
Results
Among 102 GC patients, the FGFR4-Arg388 patients showed significantly higher tumor stage (p=0.047) and worse overall survival (p=0.033) than the Gly388 patients. Immunohistochemical results showed that FGFR4-Arg388 patients were more likely to have higher vimentin (p=0.025) and p-STAT3 (p=0.009) expression compared with FGFR4-Gly388 patients. In transfected GC cells, the overexpression of FGFR4-Arg388 variant increased proliferation and invasion of GC cells, increasing resistance of GC cells to OXA compared with cells overexpressing the Gly388 allele.
Conclusion
The exploration mechanism may be through FGFR4-Arg388/STAT3/epithelial to mesenchymal transition axis regulating pivotal oncogenic properties of GC cells. The FGFR4-Arg388 variant may be a biomarker and a candidate target for adjuvant treatment of GC.

Keyword

Fibroblast growth factor receptor 4; Gly388Arg polymorphism; Gastric neoplasms; STAT3; Epithelial to mesenchymal transition

Figure

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  Fig. 1. (A) Gene sequencing including three specific fibroblast growth factor receptor 4 (FGFR4) Gly388Arg polymorphic genotype (including Gly/Gly, Gly/Arg, and Arg/Arg). (B) Significant difference was observed between patients with FGFR4-Gly388 allele and FGFR4-Arg388 variant among 102 gastric cancer (GC) patients after stratified Kaplan-Meier survival analysis. (C) Strong positive expressions of immunohistochemical markers in GC tissue were demonstrated. All H&E staining and immunohistochemical pictures were amplified 200-fold (upper) and 400-fold (lower).

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  Fig. 2. The expressions of fibroblast growth factor receptor 4 (FGFR4) mRNA and protein were illustrated in various gastric cancer (GC) cell lines and transfected cells. (A) Expressions of FGFR4 protein in different GC cell lines by western blot. (B) Expressions of FGFR4 mRNA in different GC cells by reverse transcription polymerase chain reaction (RT-PCR). (C) Expressions of FGFR4 mRNA in different GC cells by quantitative real-time polymerase chain reaction (q-PCR). Expressions of FGFR4 mRNA in SGC7901 (D) and BGC803 (E) cells (mock, negative control [NC], Gly388-transfected, and Arg388-transfected) by RT-PCR. Expressions of FGFR4 mRNA in SGC7901 (F) and BGC803 (G) cells (mock, NC, Gly388-transfected, and Arg388-transfected) by q-PCR. β-Actin was served as loading control. At least three independent detecting were performed. *p < 0.05.

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  Fig. 3. Effect of fibroblast growth factor receptor 4 (FGFR4) genotype on proliferation, invasion, and migration. (A, B) Compared with mock and negative control (NC) cells, the proliferation of Gly388- and Arg388-transfected cells was significantly enhanced. In addition, Arg388-transfected cells have a faster proliferation than Gly388-transfected cells. In addition, Arg388-transfected cells have a faster proliferation than Gly388-transfected cells. Similar results were also observed in the EdU fluorescence staining test (C-F), Transwell chambers invasion assay (G-J), and cell scratch assay (K-N). *p < 0.05. Similar results were also observed in the EdU fluorescence staining test (C-F), Transwell chambers invasion assay (G-J), and cell scratch assay (K-N). *p < 0.05.

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  Fig. 4. The relationship between fibroblast growth factor receptor 4 (FGFR4) genotype and oxaliplatin (OXA) resistance. (A-D) Effects of different OXA concentrations on the activity of SGC7901 and BGC803 cells. NC, normal control. (E-H) Flow cytometry: apoptosis rate of gastric cancer (GC) cells with different FGFR4 genotypes after OXA addition for 24 hours. (I-L) Fluorescent staining for apoptosis of GC cells with different FGFG4 genotypes (green represents early apoptosis and red represents apoptosis death). PI, propidium iodide. (M, N) Western bolt was used to detect the changes of apoptosis molecules in GC cells with different FGFG4 genotypes before and after OXA addition. *p < 0.05.

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  Fig. 5. The mechanism of fibroblast growth factor receptor 4 (FGFR4)-Arg388 influencing the oncogenic properties of gastric cancer cells. (A, B) Western blot detection of the expression of related molecules including FGFR4, signal pathway (STAT3), and epithelial to mesenchymal transition (E-cadherin and vimentin) in SGC7901 and BGC803 cells with different FGFG4 genotypes (mock, normal control [NC], Gly388, and Arg388). (C, D) The changes of related molecules between overexpression Gly388- and Arg388-transfected cells were detected, when cells were incubated with the Jak2 inhibitor AG490 to block STAT3 activation.

Reference

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