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Application of m6A demethylase FTO in inhibition of NPM1 mutant leukemia cell proliferation

A technology of leukemia cells and demethylases, applied in biochemical equipment and methods, microbiological measurement/testing, instruments, etc., to achieve the effect of enhancing the ability of cells to proliferate in vitro

Pending Publication Date: 2022-04-12
重庆医科大学国际体外诊断研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the role of m6A modification in the course of NPM1 mutant leukemia has not been reported

Method used

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  • Application of m6A demethylase FTO in inhibition of NPM1 mutant leukemia cell proliferation
  • Application of m6A demethylase FTO in inhibition of NPM1 mutant leukemia cell proliferation
  • Application of m6A demethylase FTO in inhibition of NPM1 mutant leukemia cell proliferation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. Detection of FTO expression, demethylase activity and identification of its clinical prognostic value in NPM1 mutant leukemia

[0029] 1.1 Experimental method

[0030] 1.1.1 Download and analysis of AML related data from TCGA and GEO databases

[0031] Gene expression data and clinical information of AML samples were obtained from the Cancer Genome Atlas (TCGA, http: / / www.cancergenome.nih.gov ) (n=151) and the Gene Expression Omnibus (GEO, http: / / www.ncbi.nlm.nih.gov / gds ) in the GSE14468 (n=187) and GSE15434 (n=251) datasets. RNA-seq data of AML samples were obtained from the TCGA database, and gene expression microarray data of AML samples were obtained from the GSE14468 and GSE15434 datasets, and then a t-test was used to compare the expression of FTO gene levels between NPM1-mutant AML and non-NPM1-mutant AML. difference.

[0032] 1.1.2 Culture of leukemia cells

[0033] OCI-AML3 and OCI-AML2 cells were purchased from the German Collection of Micro...

Embodiment 2

[0078] Example 2. Effect of interference with FTO expression on proliferation of NPM1 mutant leukemia cells

[0079] 2.1 Experimental method

[0080] 2.1.1 CCK-8 assay to detect cell proliferation

[0081] Collect cells in each group at 1.5 × 10 per well 4 The number of cells was seeded in 96-well plates, and 5 duplicate wells were set in each group. At the same time, a blank control was set up, and the culture was continued in the incubator; 10 μL of CCK-8 reagent was added to each well at the 0th, 24h, 48h and 72h of culture, and the mixture was mixed. Continue to culture for 3h after homogenization; detect the absorbance (OD) value at 450nm wavelength, draw the cell proliferation curve with the OD450 value as the ordinate and the culture time as the abscissa.

[0082] 2.1.2 EdU assay to detect cell proliferation

[0083] Collect cells from each group at 1.0 × 10 per well 6 The number of cells was seeded in 6-well plates and incubated with 10 μM EdU working solution at 3...

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Abstract

The invention relates to application of m6A demethylase FTO in inhibition of NPM1 mutant leukemia cell proliferation, and belongs to the technical field of biomedical research. The technical problem to be solved by the invention is to provide a new molecular target for treating NPM1 mutant leukemia. The expression level of FTO in NPM1 mutant leukemia is higher than that of non-NPM1 mutant leukemia, and high expression of FTO is related to poor prognosis of a patient; the shRNA lentiviral vector or drug meclofenac sodium is utilized to target FTO, and the growth capacity of leukemia cells can be effectively inhibited. The m6A demethylase FTO is expected to become a novel molecular treatment target and a clinical prognosis index of the NPM1 mutant leukemia.

Description

technical field [0001] The invention belongs to the technical field of biomedical research, in particular to the application of m6A demethylase FTO in inhibiting the proliferation of NPM1 mutant leukemia cells. Background technique [0002] Acute myeloid leukemia (AML) with nucleophosmin 1 (NPM1) gene mutation is the most common type of normal karyotype AML. Because of its unique clinical features, it has been regarded as a special subtype of AML. Type was included in the 2016 WHO leukemia typing guidelines. However, because the specific pathogenic mechanism has not been elucidated, the current clinical treatment has not made substantial progress. Therefore, in-depth study of the pathogenic mechanism of NPM1-mutated AML and the search for specific target molecules to achieve individualized treatment are the focus of current research. [0003] In recent years, with the development of high-throughput sequencing technology, RNA modification has become the focus of many studie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/686C12Q1/02G01N33/574
Inventor 张伶肖巧玲雷力彭美茜敬一佩任俊黄军鹏陶永红
Owner 重庆医科大学国际体外诊断研究院
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