Dataset Information

M⁶A mRNA methylation regulates CTNNB1 to promote the proliferation of hepatoblastoma.

ABSTRACT:

Background

N⁶-Methyladenosine (m⁶A) modification has been implicated in many biological processes. It is important for the regulation of messenger RNA (mRNA) stability, splicing, and translation. However, its role in cancer has not been studied in detail. Here we investigated the biological role and underlying mechanism of m⁶A modification in hepatoblastoma (HB).

Methods

We used Reverse transcription quantitative real-time PCR (RT-qPCR) and Western blotting to determine the expression of m⁶A related factors. And we clarified the effects of these factors on HB cells using cell proliferation assay, colony formation, apoptotic assay. Then we investigated of methyltransferase-like 13 (METTL3) and its correlation with clinicopathological features and used xenograft experiment to check METTL3 effect in vivo. m⁶A-Seq was used to profiled m⁶A transcriptome-wide in hepatoblastoma tumor tissue and normal tissue. Finally, methylated RNA immunoprecipitation (MeRIP) assay, RNA remaining assay to perform the regulator mechanism of MEETL3 on the target CTNNB1 in HB.

Results

In this research, we discovered that m⁶A modifications are increased in hepatoblastoma, and METTL3 is the main factor involved with aberrant m⁶A modification. We also profiled m⁶A across the whole transcriptome in hepatoblastoma tumor tissues and normal tissues. Our findings suggest that m⁶A is highly expressed in hepatoblastoma tumors. Also, m⁶A is enriched not only around the stop codon, but also around the coding sequence (CDS) region. Gene ontology analysis indicates that m⁶A mRNA methylation contributes significantly to regulate the Wnt/?-catenin pathway. Reduced m⁶A methylation can lead to a decrease in expression and stability of the CTNNB1.

Conclusion

Overall our findings suggest enhanced m⁶A mRNA methylation as an oncogenic mechanism in hepatoblastoma, METTL3 is significantly up-regulated in HB and promotes HB development. And identify CTNNB1 as a regulator of METTL3 guided m⁶A modification in HB.

SUBMITTER: Liu L

PROVIDER: S-EPMC6927193 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature

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Publications

m<sup>6</sup>A mRNA methylation regulates CTNNB1 to promote the proliferation of hepatoblastoma.

Liu Li L Wang Jing J Sun Guifeng G Wu Qiong Q Ma Ji J Zhang Xin X Huang Nan N Bian Zhixuan Z Gu Song S Xu Min M Yin Minzhi M Sun Fenyong F Pan Qiuhui Q

Molecular cancer 20191223 1

<h4>Background</h4>N<sup>6</sup>-Methyladenosine (m<sup>6</sup>A) modification has been implicated in many biological processes. It is important for the regulation of messenger RNA (mRNA) stability, splicing, and translation. However, its role in cancer has not been studied in detail. Here we investigated the biological role and underlying mechanism of m<sup>6</sup>A modification in hepatoblastoma (HB).<h4>Methods</h4>We used Reverse transcription quantitative real-time PCR (RT-qPCR) and Western ...[more]

PMID: 31870368

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Project description:Macroautophagy/autophagy is indispensable for testosterone synthesis in Leydig cells (LCs), and here we report a negative association between m6A modification and autophagy in LCs during testosterone synthesis. A gradual decrease of METTL14 (methyltransferase like 14) and an increase of ALKBH5 (alkB homolog 5, RNA demethylase) were observed in LCs during their differentiation from stem LCs to adult LCs. These events led to reduced mRNA methylation levels of N6-methyladenosine (m6A) and enhanced autophagy in LCs. Similar regulation of METTL14, ALKBH5, and m6A was also observed in LCs upon treatment with human chorionic gonadotropin (HsCG). Mechanistically, m6A modification promoted translation of PPM1A (protein phosphatase 1A, magnesium dependent, alpha isoform), a negative AMP-activated protein kinase (AMPK) regulator, but decreased expression of CAMKK2 (calcium/calmodulin-dependent protein kinase kinase 2, beta), a positive AMPK regulator, by reducing its RNA stability. Thus, m6A modification resulted in reduced AMPK activity and subsequent autophagy inhibition. We further demonstrated that ALKBH5 upregulation by HsCG was dependent on enhanced binding of the transcriptional factor CEBPB (CCAAT/enhancer binding protein [C/EBP], beta) and the TFEB (transcription factor EB) to its gene promoter. Moreover, HsCG treatment decreased METTL14 by reducing its stability. Collectively, this study highlights a vital role of m6A RNA methylation in the modulation of testosterone synthesis in LCs, providing insight into novel therapeutic strategies by exploiting m6A RNA methylation as targets for treating azoospermatism and oligospermatism patients with reduction in serum testosterone.Abbreviations: 3-MA: 3-methyladenine; ACTB: Actin, beta; ALKBH5: alkB homolog 5, RNA demethylase; AMPK: AMP-activated protein kinase; BafA1: bafilomycin A1; CAMKK2: calcium/calmodulin-dependent protein kinase kinase 2, beta; CEBPB: CCAAT/enhancer-binding protein (C/EBP), beta; ChIP: chromatin immunoprecipitation; FTO: fat mass and obesity associated; HsCG: human chorionic gonadotropin; HSD3B: 3β-hydroxysteroid dehydrogenase; LCs: Leydig cells; m6A: N6-methyladenosine; METTL14: methyltransferase like 14; METTL3: methyltransferase like 3; MTOR: mechanistic target of rapamycin kinase; PPM1A: protein phosphatase 1A, magnesium dependent, alpha isoform; PRKAA: 5'-AMP-activated protein kinase catalytic subunit alpha; SQSTM1: sequestosome 1; STK11/LKB1: serine/threonine kinase 11; TFEB: transcription factor EB; ULK1: unc-51-like kinase 1; WTAP: Wilms tumor 1-associating protein; YTHDF: YTH N6-methyladenosine RNA binding protein.

Dataset Information

M⁶A mRNA methylation regulates CTNNB1 to promote the proliferation of hepatoblastoma.

Background

Methods

Results

Conclusion

Publications

m<sup>6</sup>A mRNA methylation regulates CTNNB1 to promote the proliferation of hepatoblastoma.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Dataset Information

M6A mRNA methylation regulates CTNNB1 to promote the proliferation of hepatoblastoma.

Background

Methods

Results

Conclusion

Publications

m<sup>6</sup>A mRNA methylation regulates CTNNB1 to promote the proliferation of hepatoblastoma.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

M⁶A mRNA methylation regulates CTNNB1 to promote the proliferation of hepatoblastoma.