m(6)A Modification of lncRNA NEAT1 Regulates Chronic Myelocytic Leukemia Progression via miR-766-5p/CDKN1A Axis

Abstract

BackgroundChronic myeloid leukemia (CML) is an acquired hematopoietic stem malignant disease originating from the myeloid system. Long non-coding RNAs (lncRNAs) have been widely explored in cancer tumorigenesis. However, their roles in CML remain largely unclear.MethodsThe peripheral blood mononuclear cells (PBMCs) and CML cell lines (K562, KCL22, MEG01, BV173) were collected for in vitro research. Real-time quantitative polymerase chain reaction was used to determine the mRNA expression levels. Cell viability and apoptosis were analyzed by cell counting kit 8 and flow cytometry assays. The targeting relationships were predicted using Starbase and TargetScan and ulteriorly verified by RNA pull-down and luciferase reporter assays. Western blotting assay was performed to assess the protein expressions. N6-methyladenosine (m6A) modification sites were predicted by SRAMP and confirmed by Methylated RNA immunoprecipitation (MeRIP) assay.ResultsLncRNA nuclear-enriched abundant transcript 1 (NEAT1) expression levels were decreased in the CML cell lines and PBMCs of CML patients. Moreover, METTL3-mediated m6A modification induced the aberrant expression of NEAT1 in CML. Overexpression of NEAT1 inhibited cell viability and promoted the apoptosis of CML cells. Additionally, miR-766-5p was upregulated in CML PBMCs and abrogated the effects of NEAT1 on cell viability and apoptosis of the CML cells. Further, CDKN1A was proved to be the target gene of miR-766-5p and was downregulated in the CML PBMCs. Knockdown of CDKN1A reversed the effects of NEAT1.ConclusionThe current research elucidates a novel METTL3/NEAT1/miR-766-5p/CDKN1A axis which plays a critical role in the progression of CML.

The prognosis biomarkers based on m6A-related lncRNAs for myeloid leukemia patients

“…The result was consistent with other studies [12]. For instance, m6A modification of lncRNA NEAT1 inhibited cell viability and promoted the apoptosis of CML cells [22].…”

The Role of RNA Methyltransferase METTL3 in Normal and Malignant Hematopoiesis

“…Fang-Yi Yao et al. reported that METTL3 was downregulated in CML cells, resulting in a decrease in the protein level of nuclear enriched abundant transcript 1 ( NEAT1 ) ( 70 ). Furthermore, METTL3 downregulation in CMLs reduced its ability to modify NETA1 m 6 A, subsequently enhancing CML cell viability and inhibiting CML cell apoptosis.…”

“…Higher expression of METTL3 in AML is critical to maintain the undifferentiation of AML cells, promote the growth of AML cells, and inhibit AML cell apoptosis ( 31 , 32 , 66 ). In CML, TKI resistance always makes it more difficult to cure patients with CML, while METTL3 has been found to affect the apoptosis, proliferation, and viability of CML cells with higher expression ( 60 , 61 , 68 , 70 ). More importantly, METTL3 depletion notably damages the proliferation of primary CML cells and TKI-resistant CML cells, which suggests that METTL3 inhibitors may have novel potential to cure TKI-resistant CML patients.…”

The Role of N6-Methyladenosine (m6A) Methylation Modifications in Hematological Malignancies

“…Therefore, the findings suggest that METTL3 is a novel oncogene in the pathogenesis of CML and a potential therapeutic target for TKIs-resistant CML. Another recent finding [ 131 ] also revealed that via modulating the miR-766-5p/CDKN1A axis, METTL3 could modify long non-coding RNAs (lncRNAs) nuclear-enriched transcript 1 (NEAT1), thus controlling the progressive process of CML disorder and playing an oncogenic role in CML. NEAT1 is currently known to have low expression levels in CML cell lines or PBMCs of CML patients [ 131 ].…”

“…Another recent finding [ 131 ] also revealed that via modulating the miR-766-5p/CDKN1A axis, METTL3 could modify long non-coding RNAs (lncRNAs) nuclear-enriched transcript 1 (NEAT1), thus controlling the progressive process of CML disorder and playing an oncogenic role in CML. NEAT1 is currently known to have low expression levels in CML cell lines or PBMCs of CML patients [ 131 ]. Mechanistically, the dysfunction of m 6 A methyltransferase METTL3 causes the abnormal expression of NEAT1 in CML, and the overexpression of NEAT1 greatly promotes the death of CML cells.…”