Abstract
Background
Frizzled (FZD) proteins function as receptors for WNT ligands. Members in FZD family including FZD2, FZD4, FZD7, FZD8 and FZD10 have been demonstrated to mediate cancer cell epithelial-mesenchymal transition (EMT).
Methods
CCLE and TCGA databases were interrogated to reveal the association of FZD5 with EMT. EMT was analyzed by investigating the alterations in CDH1 (E-cadherin), VIM (Vimentin) and ZEB1 expression, cell migration and cell morphology. Transcriptional modulation was determined by ChIP in combination with Real-time PCR. Survival was analyzed by Kaplan–Meier method.
Results
In contrast to other FZDs, FZD5 was identified to prevent EMT in gastric cancer. FZD5 maintains epithelial-like phenotype and is negatively modulated by transcription factors SNAI2 and TEAD1. Epithelial-specific factor ELF3 is a downstream effecter, and protein kinase C (PKC) links FZD5 to ELF3. ELF3 represses ZEB1 expression, further guarding against EMT. Moreover, FZD5 signaling requires its co-receptor LRP5 and WNT7B is a putative ligand for FZD5. FZD5 and ELF3 are associated with longer survival, whereas SNAI2 and TEAD1 are associated with shorter survival.
Conclusions
Taken together, FZD5-ELF3 signaling blocks EMT, and plays a potential tumor-suppressing role in gastric cancer.
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Background
Frizzled (FZD) proteins are G protein-coupled receptors (GPCRs). The extracellular N-terminus contains a cysteine-rich domain (CRD) through which FZDs bind WNT ligands. The intracellular C-terminus binds the PDZ domain of Dishevelled (Dvl) and interacts with G proteins. Up to now, 10 FZDs have been identified in human. These receptors mediate β-catenin pathway and various β-catenin-independent pathways depending on cellular context. WNT/β-catenin pathway is oncogenic and involved in almost every aspect of tumor development. In addition, β-catenin-independent WNT/PCP and WNT/Ca2+-PKC pathways are also implicated in tumor progression [1,2,3,4]. In recent years, some novel pathways such as WNT/Stat3 and WNT/Yap-Taz have been successively identified [5,6,7,8], indicating that FZD signalings are far more complicated and still incompletely comprehended.
Epithelial-mesenchymal transition (EMT) in tumor cells endows them with enhanced motility thereby increasing their metastatic potentiality. This process is characterized by reduced expression of epithelial-related factors and increased expression of mesenchymal-related factors. Mechanistically, EMT is driven by some transcription factors such as SNAI1/2 and ZEB1/2. Studies have shown that FZD signalings induce tumor cell EMT and metastasis. FZD4, FZD7 and FZD10 promote EMT through β-catenin pathway in prostate, liver and breast cancer [9,10,11]. FZD2 mediates WNT5-induced EMT dependent on Stat3 signaling in liver, lung, breast and colon cancer [5], and FZD8 mediates WNT11-induced EMT by interacting with TGF-β signaling in prostate cancer [12].
In gastric cancer, FZD4 and FZD7 similarly promote EMT and metastasis by activating β-catenin pathway [13, 14]. Genetic deletion of FZD7 was shown to inhibit the growth of gastric adenomas in vivo [15]. FZD2 and FZD8, through β-catenin pathway but not β-catenin-independent pathways, induce gastric cancer cell proliferation, migration, invasion and metastasis [16, 31]. Actually, WNT7A functions as an EMT inhibitor in lung cancer [32]. Moreover, interrogation of CCLE database showed that FZD5 is also associated with epithelial-like phenotype in lung cancer (data not shown).
As shown in our study, FZD5 is transcriptional inhibited by TEAD1-SNAI2 complex. Activation of tumor-suppressing Hippo pathway results in phosphorylation and degradation of Yap and Taz. When Yap and Taz are not phosphorylated, they translocate into the nucleus to bind transcription factor TEAD. Yap/Taz-TEAD signaling induces gastric cancer cell EMT, stemness, drug resistance and metastasis [33,41, 42], and promotes prostate cancer progression by interacting with NFκB [43]. In gastric cancer, ELF3 maintains epithelial-like phenotype, prevents EMT and is associated with longer survival, acting as a putative tumor suppressor. ELF3 also inhibits EMT in ovarian and breast cancer cells [44, 45]. Mechanistically, ELF3 represses activity and expression of EMT transcription factor ZEB1 [46, 47].
Activation of β-catenin pathway requires not only FZDs, but also their co-receptors LRP5/6 which is not involved in non-canonical β-catenin-independent pathway. A recent study reported that FZDs-LRP5/6 interaction can initiate β-catenin pathway in the absence of WNTs [48]. Surprisingly, another study revealed that FZDs-LRP5/6 interaction can block activation of FZD-mediated non-canonical pathway, further preventing tumor metastasis [49]. These findings, in combination with our observations, suggest that FZDs-LRP5/6 interaction may act as a tumor promoter by activating β-catenin pathway or a tumor suppressor by antagonizing β-catenin-independent pathway, depending on tumor or cell context.
Conclusions
In summary, our study for the first time uncovers that FZD5-ELF3 signaling prevents EMT and is associated with favorable prognosis in gastric cancer. Therefore, FZD5 and ELF3 function as putative tumor suppressors in this type of cancer. Antibodies against FZD5 including ipafricept and vantictumab have been shown effective on certain cancer types [50]. However, our study warns out that targeting FZD5 should be cautious and tumor-specific, since treatment with these antibodies may potentially promote metastasis in some other types of cancers including gastric cancer.
Availability of data and materials
All data generated or analysed during this study are included in this published article.
Abbreviations
- FZD:
-
Frizzled
- EMT:
-
Epithelial-mesenchymal transition
- PKC:
-
Protein kinase C
- GPCRs:
-
G protein-coupled receptors
- CCLE:
-
Cancer Cell Line Encyclopedia
- TCGA:
-
The Cancer Genome Atlas
- ChIP:
-
Chromatin Immunoprecipitation
- CoIP:
-
Co-immunoprecipitation
- LRP5:
-
LDL receptor-related proteins 5
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This work is supported by two grants from Department of Science and Technology of Liaoning Province (2019-MS-363, 2017225028) and a project funded by China Postdoctoral Science Foundation (2020M681014).
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DD, LN and KZ performed the experiments, analyzed the data and wrote the manuscript. ZW, YS, QZ and JG performed the experiments. WW and CZ designed the project, interpreted the data, and drafted the manuscript. All authors read and approved the final manuscript.
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Gastric cancer tissues were obtained from Sheng**g Hospital China Medical University with the informed consent of the patients. Institutional Research Ethics Committee of China Medical University approved the use of these tissues for research purposes.
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Dong, D., Na, L., Zhou, K. et al. FZD5 prevents epithelial-mesenchymal transition in gastric cancer. Cell Commun Signal 19, 21 (2021). https://doi.org/10.1186/s12964-021-00708-z
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DOI: https://doi.org/10.1186/s12964-021-00708-z