m6A mRNA methylation regulates CTNNB1 to promote the proliferation of hepatoblastoma

Abstract

BackgroundN6-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).MethodsWe 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.ResultsIn 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/β-catenin pathway. Reduced m6A methylation can lead to a decrease in expression and stability of the CTNNB1.ConclusionOverall 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.

YTHDC1 gene polymorphisms and hepatoblastoma susceptibility in Chinese children: A seven‐center case–control study

“…36 Growing evidence shows that m 6 A contributes to the progression of cancer. Liu et al 37 demonstrated that enhanced m 6 A mRNA methylation plays an oncogenic mechanism role in hepatoblastoma. Yang et al 38 revealed that YTHDF2 was overexpressed in hepatocellular carcinoma cells and miR-145 modulates m 6 A levels by targeting the 3 0 -UTR of YTHDF2 mRNA in hepatocellular carcinoma cells.…”

m6A RNA methylation impacts fate choices during skin morphogenesis

“…Recent studies on other cell types and cancers have also reported connections between WNT signaling and m6A levels (Bai Y et al, 2019;Miao W et al, 2019;Han et al, 2020;Liu et al, 2019;Miao et al, 2019;Tang et al, 2020;Zhang et al, 2019). In some of these cases, m6A loss resulted in an elevation of WNT-signaling, while in others, it was diminished, and a variety of mechanisms have been proposed to explain these seemingly context-dependent effects.…”

The prognostic value of METTL3 in cancer patients: a meta-analysis

“…METTL3 was considered to be a methyltransferase, part of the methyltransferase complex, and was responsible for the m6A modi cation [27,53].METTL3 was involved in many signal pathways such as PI3K / Akt [31,[54][55][56][57][58], MAPK [22] ,Wnt /beta-Catenin [47,59,60] and p38 / ERK [61]pathways,which were all associated with tumor deterioration. In addition, METTL3 could in uence the development of tumor by regulating some transcription factors or important oncogenes.Studies had found that METTL3 could positively regulate the expression of oncogene EZH2 [24,25,62].It promoted the expression of MYC as well as increased stability of protein by regulating the m6A methylation of MYC mRNA to lead to carcinogenesis in PCA and gastric cancer, promote the occurrence of OSCC tumor and affect the growth and invasion of BCA cells [40,[63][64][65][66].When METTL3 was silenced, it inhibited the activity of the Wnt pathway by reducing the m6A methylation level of LEF1 mRNA and reducing protein expression [59,67].…”