[The Regulatory Effect of RNA m A Methylation Modification on Gene Expression in t (8;21) AML Cells by MeRIP-qPCR].

Objective: To confirm the direct regulatory effect of -mediated RNA mA modification on the gene in t (8;21) acute myeloid leukemia (AML) cells through MeRIP combined with reverse transcription real-time quantitative PCR (RT-qPCR) technology.

Methods: The lentivirus-mediated shRNA target or gene was used to transfect the t (8;21) AML cell lines: Kasumi-1 and SKNO-1, and cells transfected with randomly shuffled shRNA as the control. Using the Ultrapure RNA Extraction Kit (DNase I) to extract RNA.

HIF1α-mediated transactivation of WTAP promotes AML cell proliferation via mA-dependent stabilization of KDM4B mRNA.

Hypoxia inducible factor 1α (HIF1α) is abnormally overexpressed in t(8;21) acute myeloid leukemia (AML) and functions as an oncogene through transactivating DNA methyltransferase 3 alpha leading to DNA hypermethylation. However, it remains unclear whether HIF1α influences RNA N-methyladenosine (mA) methyltransferases. Here, we show that HIF1α promotes the expression of Wilms tumor 1-associated protein (WTAP), a main component of the mA methyltransferase complex, markedly alters the transcriptome-wide mA distribution and enhances cell proliferation in t(8;21) AML.

[The Relationship between HIF1α and WTAP Expression Level in t(8;21) Acute Myeloid Leukemia].

Objective: To investigate the relationship between hypoxia inducible factor 1 (HIF1α) and Wilms' tumor 1associating protein (WTAP) expression level in t(8;21) acute myeloid leukemia cells.

Methods: The t(8;21) acute myeloid leukemia cell lines, including SKNO-1 and Kasumi-1 were treated by Echinomycin for 24 h, RT-qPCR and Western blot were used to detect the expression levels of WTAP mRNA and the protein. The CoCl was used to induce the hypoxia of the cells for 24 h, the expression levels of HIF1α, WTAP protein were detected by Western blot.

YTHDF2 is a potential target of AML1/ETO-HIF1α loop-mediated cell proliferation in t(8;21) AML.

The t(8;21) fusion product, AML1/ETO, and hypoxia-inducible factor 1α (HIF1α) form a feed-forward transcription loop that cooperatively transactivates the DNA methyltransferase 3a gene promoter that leads to DNA hypermethylation and drives leukemia cell growth. Suppression of the RNA N-methyladenosine (mA)-reader enzyme YTH N-methyladenosine RNA binding protein 2 (YTHDF2) specifically compromises cancer stem cells in acute myeloid leukemia (AML) but promotes hematopoietic stem cell expansion without derailing normal hematopoiesis. However, the relevance of expression between AML1/ETO-HIF1α loop and YTHDF2, and its functional relationship with t(8;21) AML have not been documented.