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Research
FTO demethylates m6A modifications in HOXB13 mRNA and promotes endometrial cancer metastasis by activating the WNT signaling pathway
Jinghe Lang
Chinese Academy of Medical Sciences and Peking Union Medical College
Corresponding Author
ORCiD: https://orcid.org/0000-0002-5087-3788
Wenjun Cheng
Nanjing Medical University First Affiliated Hospital
Corresponding Author
Lan Zhu
Chinese Academy of Medical Sciences & Peking Union Medical College Hospital & peking union medical college
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background Abnormal FTO expression causes faulty m6A modifications in mRNA and affects tumor progression. However, the expression and function of FTO in endometrial cancer (EC) and the genes regulated by FTO remain unclear.
Methods qPCR detected the FTO expression level in EC tissues, and IHC staining determined the FTO protein expression in tissue arrays. FTO was stably overexpressed or knocked down in EC cell lines by lentiviruses. The biological roles of FTO in cancer cell metastasis and invasion were evaluated by wound-healing, Transwell assays and mouse intra-abdominal implantation models. mRNA-seq, RIP-seq, and MeRIP-seq were combined to comprehensively map the genes regulated by FTO. RNA pull-down, dual-luciferase reporter and RNA stability experiments confirmed that YTHDF2 regulates the metabolism of HOXB13 mRNA via m6A. Western blot analysis confirmed that HOXB13 regulates EC cell metastasis and invasion through the WNT signaling pathway.
Results FTO is more highly expressed in metastatic EC and can promote tumor metastasis and invasion in vivo and in vitro. By removing the m6A modification in the 3'UTR region of HOXB13, FTO abolishes m6A recognition by YTHDF2 and promotes the stability of HOXB13 mRNA and protein expression. High HOXB13 expression activates the WNT signaling pathway, promotes c-myc, snail, MMP2, MMP7, and MMP9 expression, and inhibits E-cadherin expression, leading to tumor metastasis and invasion.
Conclusions FTO promotes HOXB13 expression through m6A, activates the WNT signaling pathway, and promotes EC invasion and metastasis. FTO is a new potential target for the treatment of tumor metastasis.
Keywords
Endometrial cancer, FTO, HOXB13, m6A, Cancer metastasis
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- Additionalfile2TableS2.docx
- Additionalfile5TableS5mRNAseqdata.xlsx
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- Additionalfile6TableS6RIPseqdata.xlsx
- Additionalfile2TableS2.docx
- Additionalfile1TableS1.docx
- Additionalfile8FigureS1.tif
- Additionalfile7TableS7MeRIPseqdata.xlsx
- Additionalfile3TableS3.docx
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- Additionalfile6TableS6RIPseqdata.xlsx
- Additionalfile4TableS4.docx
- Additionalfile8FigureS1.tif
- Additionalfile7TableS7MeRIPseqdata.xlsx
- Additionalfile5TableS5mRNAseqdata.xlsx
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Research
FTO demethylates m6A modifications in HOXB13 mRNA and promotes endometrial cancer metastasis by activating the WNT signaling pathway
Jinghe Lang
Chinese Academy of Medical Sciences and Peking Union Medical College
Corresponding Author
ORCiD: https://orcid.org/0000-0002-5087-3788
Wenjun Cheng
Nanjing Medical University First Affiliated Hospital
Corresponding Author
Lan Zhu
Chinese Academy of Medical Sciences & Peking Union Medical College Hospital & peking union medical college
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 26 May, 2020
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background Abnormal FTO expression causes faulty m6A modifications in mRNA and affects tumor progression. However, the expression and function of FTO in endometrial cancer (EC) and the genes regulated by FTO remain unclear.
Methods qPCR detected the FTO expression level in EC tissues, and IHC staining determined the FTO protein expression in tissue arrays. FTO was stably overexpressed or knocked down in EC cell lines by lentiviruses. The biological roles of FTO in cancer cell metastasis and invasion were evaluated by wound-healing, Transwell assays and mouse intra-abdominal implantation models. mRNA-seq, RIP-seq, and MeRIP-seq were combined to comprehensively map the genes regulated by FTO. RNA pull-down, dual-luciferase reporter and RNA stability experiments confirmed that YTHDF2 regulates the metabolism of HOXB13 mRNA via m6A. Western blot analysis confirmed that HOXB13 regulates EC cell metastasis and invasion through the WNT signaling pathway.
Results FTO is more highly expressed in metastatic EC and can promote tumor metastasis and invasion in vivo and in vitro. By removing the m6A modification in the 3'UTR region of HOXB13, FTO abolishes m6A recognition by YTHDF2 and promotes the stability of HOXB13 mRNA and protein expression. High HOXB13 expression activates the WNT signaling pathway, promotes c-myc, snail, MMP2, MMP7, and MMP9 expression, and inhibits E-cadherin expression, leading to tumor metastasis and invasion.
Conclusions FTO promotes HOXB13 expression through m6A, activates the WNT signaling pathway, and promotes EC invasion and metastasis. FTO is a new potential target for the treatment of tumor metastasis.
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