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Title:
WTAP facilitates progression of hepatocellular carcinoma via m6A-HuR-dependent epigenetic silencing of ETS1 | Molecular Cancer
Description:
Background N6-methyladenosine (m6A) methylation, a well-known modification with new epigenetic functions, has been reported to participate in the tumorigenesis of hepatocellular carcinoma (HCC), providing novel insights into the molecular pathogenesis of this disease. However, as the key component of m6A methylation, Wilms tumor 1-associated protein (WTAP) has not been well studied in HCC. Here we investigated the biological role and underlying mechanism of WTAP in liver cancer. Methods We determined the expression of WTAP and its correlation with clinicopathological features using tissue microarrays and the Cancer Genome Atlas (TCGA) dataset. And we clarified the effects of WTAP on HCC cells using cell proliferation assay, colony formation, Edu assay and subcutaneous xenograft experiments. We then applied RNA sequencing combined with gene expression omnibus (GEO) data to screen candidate targets of WTAP. Finally, we investigated the regulatory mechanism of WTAP in HCC by m6A dot blot assay, methylated RNA immunoprecipitation (MeRIP) assay, dual luciferase reporter assay, RNA immunoprecipitation (RIP) assay and Chromatin immunoprecipitation (ChIP) assay. Results We demonstrated that WTAP was highly expressed in HCC which indicated the poor prognosis, and that WTAP expression served as an independent predictor of HCC survival. Functionally, WTAP promoted the proliferation capability and tumor growth of HCC cells in vitro and in vivo. Furthermore, ETS proto-oncogene 1 (ETS1) was identified as the downstream effector of WTAP. The m6A modification regulated by WTAP led to post-transcriptional suppression of ETS1, with the implication of Hu-Antigen R (HuR) as an RNA stabilizer. Then ETS1 was found to inhibit the progression of HCC and could rescue the phenotype induced by WTAP deficiency. Moreover, WTAP modulated the G2/M phase of HCC cells through a p21/p27-dependent pattern mediated by ETS1. Conclusion We have identified that WTAP is significantly up-regulated in HCC and promotes liver cancer development. WTAP-guided m6A modification contributes to the progression of HCC via the HuR-ETS1-p21/p27 axis. Our study is the first to report that WTAP-mediated m6A methylation has a crucial role in HCC oncogenesis, and highlights WTAP as a potential therapeutic target of HCC treatment.
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Keywords {🔍}
wtap, ets, hcc, cells, cell, expression, rna, article, additional, file, fig, tumor, google, scholar, figure, cas, protein, assay, knockdown, data, analysis, mrna, cancer, hur, methylation, tissues, modification, huh, level, china, silencing, cycle, chen, proliferation, survival, mettl, wang, liu, role, carcinoma, based, usa, staining, rtqpcr, kit, performed, regulation, overexpression, hcclm, control,
Topics {✒️}
ras/raf/mek/erk pathway [48] hrp-conjugated anti-mouse immunoglobulin n6-methyladenosine-dependent regulation p21/p27-dependent pattern mediated m6a-hur-dependent epigenetic silencing full size image pre-cooled ripa buffer xist rna-mediated silencing ice-cold 20ssc buffer rna n6-methyladenosine methyltransferase rna n6-methyladenosine methyltransferase article download pdf dual-glo luciferase system post-transcriptional gene regulation inducible p21waf1/cip1 transcription hrp-conjugated secondary antibodies wtap-wt1-tbl1 axis metastasis-related markers mmp7 immunoprecipitated rna-protein complex m6a-modified pri-mir126 m6a-hur-dependent pathway ythdf2-dependent posttranscriptional silencing pre-coated diluted matrigel combined multi-organ transplantation rna n6-methyladenosine kaplan-meier analysis revealed hur-ets1-p21/p27 axis distinct cis-acting elements m6a-modified transcripts relied wtap-intermediated m6a methylation p21/p27-dependent pattern m6a-hur mediated pattern patient-derived tumor xenograft wtap-guided m6a modification wtap-m6a-ythdf2-ets2 pathway wtap-related m6a methylation wtap-mediated m6a methylation magnetic beads pre-coated kaplan-meier analysis based mammalian rna demethylase n6-methyladenosine merip fundamental research funds rna-sequencing rt-qpcr wt1-tbl1 axis [45] wild-type reporters embodied wtap-mediated m6a modification privacy choices/manage cookies post-transcriptional suppression ythdf3 facilitates translation tumor size measurement
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mainEntity:
headline:WTAP facilitates progression of hepatocellular carcinoma via m6A-HuR-dependent epigenetic silencing of ETS1
description:N6-methyladenosine (m6A) methylation, a well-known modification with new epigenetic functions, has been reported to participate in the tumorigenesis of hepatocellular carcinoma (HCC), providing novel insights into the molecular pathogenesis of this disease. However, as the key component of m6A methylation, Wilms tumor 1-associated protein (WTAP) has not been well studied in HCC. Here we investigated the biological role and underlying mechanism of WTAP in liver cancer. We determined the expression of WTAP and its correlation with clinicopathological features using tissue microarrays and the Cancer Genome Atlas (TCGA) dataset. And we clarified the effects of WTAP on HCC cells using cell proliferation assay, colony formation, Edu assay and subcutaneous xenograft experiments. We then applied RNA sequencing combined with gene expression omnibus (GEO) data to screen candidate targets of WTAP. Finally, we investigated the regulatory mechanism of WTAP in HCC by m6A dot blot assay, methylated RNA immunoprecipitation (MeRIP) assay, dual luciferase reporter assay, RNA immunoprecipitation (RIP) assay and Chromatin immunoprecipitation (ChIP) assay. We demonstrated that WTAP was highly expressed in HCC which indicated the poor prognosis, and that WTAP expression served as an independent predictor of HCC survival. Functionally, WTAP promoted the proliferation capability and tumor growth of HCC cells in vitro and in vivo. Furthermore, ETS proto-oncogene 1 (ETS1) was identified as the downstream effector of WTAP. The m6A modification regulated by WTAP led to post-transcriptional suppression of ETS1, with the implication of Hu-Antigen R (HuR) as an RNA stabilizer. Then ETS1 was found to inhibit the progression of HCC and could rescue the phenotype induced by WTAP deficiency. Moreover, WTAP modulated the G2/M phase of HCC cells through a p21/p27-dependent pattern mediated by ETS1. We have identified that WTAP is significantly up-regulated in HCC and promotes liver cancer development. WTAP-guided m6A modification contributes to the progression of HCC via the HuR-ETS1-p21/p27 axis. Our study is the first to report that WTAP-mediated m6A methylation has a crucial role in HCC oncogenesis, and highlights WTAP as a potential therapeutic target of HCC treatment.
datePublished:2019-08-22T00:00:00Z
dateModified:2019-08-22T00:00:00Z
pageStart:1
pageEnd:19
license:http://creativecommons.org/publicdomain/zero/1.0/
sameAs:https://doi.org/10.1186/s12943-019-1053-8
keywords:
N6-methyladenosine (m6A)
Wilms tumor 1-associated protein (WTAP)
Hepatocellular carcinoma (HCC)
ETS1
Cancer Research
Oncology
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headline:WTAP facilitates progression of hepatocellular carcinoma via m6A-HuR-dependent epigenetic silencing of ETS1
description:N6-methyladenosine (m6A) methylation, a well-known modification with new epigenetic functions, has been reported to participate in the tumorigenesis of hepatocellular carcinoma (HCC), providing novel insights into the molecular pathogenesis of this disease. However, as the key component of m6A methylation, Wilms tumor 1-associated protein (WTAP) has not been well studied in HCC. Here we investigated the biological role and underlying mechanism of WTAP in liver cancer. We determined the expression of WTAP and its correlation with clinicopathological features using tissue microarrays and the Cancer Genome Atlas (TCGA) dataset. And we clarified the effects of WTAP on HCC cells using cell proliferation assay, colony formation, Edu assay and subcutaneous xenograft experiments. We then applied RNA sequencing combined with gene expression omnibus (GEO) data to screen candidate targets of WTAP. Finally, we investigated the regulatory mechanism of WTAP in HCC by m6A dot blot assay, methylated RNA immunoprecipitation (MeRIP) assay, dual luciferase reporter assay, RNA immunoprecipitation (RIP) assay and Chromatin immunoprecipitation (ChIP) assay. We demonstrated that WTAP was highly expressed in HCC which indicated the poor prognosis, and that WTAP expression served as an independent predictor of HCC survival. Functionally, WTAP promoted the proliferation capability and tumor growth of HCC cells in vitro and in vivo. Furthermore, ETS proto-oncogene 1 (ETS1) was identified as the downstream effector of WTAP. The m6A modification regulated by WTAP led to post-transcriptional suppression of ETS1, with the implication of Hu-Antigen R (HuR) as an RNA stabilizer. Then ETS1 was found to inhibit the progression of HCC and could rescue the phenotype induced by WTAP deficiency. Moreover, WTAP modulated the G2/M phase of HCC cells through a p21/p27-dependent pattern mediated by ETS1. We have identified that WTAP is significantly up-regulated in HCC and promotes liver cancer development. WTAP-guided m6A modification contributes to the progression of HCC via the HuR-ETS1-p21/p27 axis. Our study is the first to report that WTAP-mediated m6A methylation has a crucial role in HCC oncogenesis, and highlights WTAP as a potential therapeutic target of HCC treatment.
datePublished:2019-08-22T00:00:00Z
dateModified:2019-08-22T00:00:00Z
pageStart:1
pageEnd:19
license:http://creativecommons.org/publicdomain/zero/1.0/
sameAs:https://doi.org/10.1186/s12943-019-1053-8
keywords:
N6-methyladenosine (m6A)
Wilms tumor 1-associated protein (WTAP)
Hepatocellular carcinoma (HCC)
ETS1
Cancer Research
Oncology
image:
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author:
name:Yunhao Chen
affiliation:
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