Anti-tumor activity of a recombinant soluble Fzd7 decoy receptor in human gastric and colon cancer cells

Document Type : Original Article


1 Department of Immunology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran

2 Molecular and Cell Biology Research Center, Mazandaran University of Medical Sciences, Sari, Iran

3 Department of Infectious Diseases, Antimicrobial Resistance Research Center, Mazandaran University of Medical Sciences, Sari, Iran


Objective(s): Frizzled-7, the most common receptor of the Wnt signaling pathway, was significantly over-expressed in gastric (GC) and colorectal (CRC) cancers and stimulated tumorigenesis. The extracellular domain of Fzd7 (sFzd7) as a decoy receptor, could competitively bound with ligands and antagonize the interaction between Fzd7 receptors and Wnt ligands. 
Materials and Methods: We expressed and purified the extracellular region of Fzd7 including cysteine-rich domain (33 aa–185 aa) from Escherichia coli by chromatography. The effect of sFzd7 was evaluated on AGS gastric and SW480 colon cancer cell lines expressing high levels of Fzd7 receptor. Accordingly, cell viability and apoptosis were measured using MTT and flow cytometry assays, respectively. Real-Time PCR determined the relative expression of the β-catenin and cyclin-D1 genes. 
Results: After three days of treatment with sFzd7, the viability of AGS and SW480 cell lines was decreased in a dose-dependent manner. In addition, sFzd7 at concentrations of 10 and 20 ug/ml increased the rate of apoptosis. Especially at the concentration of 20 ug/ml, the apoptosis rate was remarkably high in AGS (P-value= 0.003) and SW480 cells (P-value= 0.0007). Finally, the expressions of β-catenin (P-value= 0.01) and cyclin-D1 (P-value= 0.02) were obviously decreased in SW480 cells. The same results were obtained in AGS cells, although not statistically significant. 
Conclusion: sFzd7 decoy receptor inhibits tumor cell progression by attenuating the Wnt pathway through inhibiting Fzd7 receptors and Wnt ligand interaction. Hence, sFzd7 can be proposed as a candidate therapy for GC and CRC cells with high levels of Fzd7 expression.


1. Arnold M, Abnet CC, Neale RE, Vignat J, Giovannucci EL, McGlynn KA, et al. Global burden of 5 major types of gastrointestinal cancer. Gastroenterology 2020; 159:335-349. e315.
2. Thrift AP, El-Serag HB. Burden of gastric cancer. Clin Gastroenterol Hepatol 2020; 18:534-542.
3. Wang L-Y, Zhao S, Lv G-J, Ma X-J, Zhang J-B. Mechanisms of resveratrol in the prevention and treatment of gastrointestinal cancer. World J. Clin. Cases 2020; 8:2425-2437.
4. Kim MJ, Huang Y, Park J-I. Targeting wnt signaling for gastrointestinal cancer therapy: Present and evolving views. Cancers 2020; 12:3638-3665.
5. Zhan T, Rindtorff N, Boutros M. Wnt signaling in cancer. Oncogene 2017; 36:1461-1473.
6. Sherwood V. WNT signaling: an emerging mediator of cancer cell metabolism? Mol Cell Biol 2015; 35:2-10.
7. Dijksterhuis JP, Baljinnyam B, Stanger K, Sercan HO, Ji Y, Andres O, et al. Systematic mapping of WNT-FZD protein interactions reveals functional selectivity by distinct WNT-FZD pairs. J Biol Chem 2015; 290:6789-6798.
8. Phesse T, Flanagan D, Vincan E. Frizzled7: a promising Achilles’ heel for targeting the Wnt receptor complex to treat cancer. Cancers 2016; 8:50-82.
9. Li G, Su Q, Liu H, Wang D, Zhang W, Lu Z, et al. Frizzled7 promotes epithelial-to-mesenchymal transition and stemness via activating canonical Wnt/β-catenin pathway in gastric cancer. Int J Biol Sci 2018; 14:280-293.
10. Jothimani M, Loganathan L, Palanisamy P, Muthusamy K. Regulatory pathways of colorectal cancer and their synergistic cross-talk mechanism. Ann colorectal res 2020; 8:105-119.
11. Ye C, Xu M, Lin M, Zhang Y, Zheng X, Sun Y, et al. Overexpression of FZD7 is associated with poor survival in patients with colon cancer. Pathol Res Pract 2019; 215:152478.
12. Cheng Y, Li L, Pan S, Jiang H, Jin H. Targeting frizzled-7 decreases stemness and chemotherapeutic resistance in gastric cancer cells by suppressing Myc expression. Med Sci Monit 2019; 25:8637-8644.
13. Ueno K, Hazama S, Mitomori S, Nishioka M, Suehiro Y, Hirata H, et al. Down-regulation of frizzled-7 expression decreases survival, invasion and metastatic capabilities of colon cancer cells. Br J Cancer 2009; 101:1374-1381.
14. Xie W, Zhang Y, He Y, Zhang K, Wan G, Huang Y, et al. A novel recombinant human Frizzled-7 protein exhibits anti-tumor activity against triple negative breast cancer via abating Wnt/β-catenin pathway. Int J Biochem Cell Biol 2018; 103:45-55.
15. Schmuck R, Warneke V, Behrens H-M, Simon E, Weichert W, Röcken C. Genotypic and phenotypic characterization of side population of gastric cancer cell lines. Am J Clin Pathol 2011; 178:1792-1804.
16. Ueno K, Hiura M, Suehiro Y, Hazama S, Hirata H, Oka M, et al. Frizzled-7 as a potential therapeutic target in colorectal cancer. Neoplasia 2008; 10:697-705.
17. Konner J, Dupont J. Use of soluble recombinant decoy receptor vascular endothelial growth factor trap (VEGF Trap) to inhibit vascular endothelial growth factor activity. Clin Colorectal Cancer 2004; 4:S81-S85.
18. de Moura PR, Watanabe L, Bleicher L, Colau D, Dumoutier L, Lemaire MM, et al. Crystal structure of a soluble decoy receptor IL-22BP bound to interleukin-22. FEBS Lett 2009; 583:1072-1077.
19. Itatani Y, Kawada K, Yamamoto T, Sakai Y. Resistance to anti-angiogenic therapy in cancer—alterations to anti-VEGF pathway. Int J Mol Sci 2018; 19:1232-1249.
20. Ahmadzadeh M, Farshdari F, Nematollahi L, Behdani M, Mohit E. Anti-HER2 scFv expression in Escherichia coli SHuffle® T7 express cells: effects on solubility and biological activity. Mol Biotechnol 2020; 62:18-30.
21. Li Y-J, Wei Z-M, Meng Y-X, Ji X-R. β-catenin up-regulates the expression of cyclinD1, c-myc and MMP-7 in human pancreatic cancer: relationships with carcinogenesis and metastasis. World J Gastroenterol 2005; 11:2117-2123.
22. Marcolino TF, Pimenta CAM, Neto RA, Castelo P, Silva MS, Forones NM, et al. p53, Cyclin-D1, β-catenin, APC and c-myc in tumor tissue from colorectal and gastric cancer patients with suspected Lynch syndrome by the Bethesda criteria. APJCP 2020; 21:343.
23. Ripple MJ, Parker Struckhoff A, Trillo-Tinoco J, Li L, Margolin DA, McGoey R, et al. Activation of c-Myc and Cyclin D1 by JCV T-Antigen and β-catenin in colon cancer. PLoS One 2014; 9:e106257.
24. DeAlmeida VI, Miao L, Ernst JA, Koeppen H, Polakis P, Rubinfeld B. The soluble wnt receptor Frizzled8CRD-hFc inhibits the growth of teratocarcinomas in vivo. Cancer Res 2007; 67:5371-5379.
25. Wei W, Chua M-S, Grepper S, So SK. Soluble Frizzled-7 receptor inhibits Wnt signaling and sensitizes hepatocellular carcinoma cells towards doxorubicin. Mol Cancer 2011; 10:1-12.