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Application of METTL3 in AML chemotherapy drug resistance

A drug-resistant and versatile technology, applied in the fields of molecular biology and medicine, to achieve the effects of strong specificity, simple detection, and improved diagnostic accuracy

Pending Publication Date: 2022-02-11
SHANDONG UNIV QILU HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current AML chemotherapy resistance still needs further research

Method used

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  • Application of METTL3 in AML chemotherapy drug resistance
  • Application of METTL3 in AML chemotherapy drug resistance
  • Application of METTL3 in AML chemotherapy drug resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Detection of Mettl3 genes and association analysis of clinical chemical drug treatment and prognostic evaluation of patients with acute myeloid leukemia

[0063] 1.1 research object

[0064] 37 patients with acute myeloid leukemia were selected, September to September 2019, in September 2019, the treatment of Qilu Hospital, Shandong University, clinical data obtained from the medical record. Most subjects are Han residents in Shandong Province. 3 ml of the test personnel was taken, and a single nuclear cell was separated. All subjects were requested by the Ethics Committee of the Qilu Hospital of Shandong University.

[0065] 1.2 RNA extraction

[0066] 1 1 mltrizol was added to the cells after PBS wash, and the gun head was blown to mix until the sample was completely dissolved, and the sample volume should not exceed 10% of Trizol volume.

[0067] 2 0.2 ml of chloroform is added to each 1mltrizol lysate, and the upper and lower reverse mix is ​​mixed 15s, and th...

Embodiment 2

[0085] Example 2 Acute myeloid leukemia patient chemical therapeutic reaction and prognostic evaluation test kit

[0086] Since Mettl3 is significantly correlated with the clinical chemotherapy response and prognosis of patients with acute myeloid leukemia, the METTL3 gene-specific primer can be used to amplify detection by the patient's cDNA as a template.

[0087] Preparation of a kit (100), which contains the material composition as shown in Table 2:

[0088]Table 2 kit composition

[0089]

[0090] The subject bone marrow was extracted, and a single nuclear cell expressing CD34 was isolated, and RNA was extracted in conventional methods. The PCR reaction was carried out by chemical treatment reaction and prognostic evaluation test kit using acute myeloid leukemia patients, and the reaction product was sequenced, and the sequencing results were tested and analyzed using BLAST software while quantitative detection. The results of a separate construction verification set (3 cas...

Embodiment 3

[0091] Example 3 Effect of Mettl3 on the drug sensitive model of humanized AML mice

[0092] In order to clarify METTL3 as the feasibility of AML chemotherapy drug resistance to the treatment of targets, this example uses lentiviral transfection techniques to express METTL3 and METTL3 methyltransferase functional mutants Mettl3-CD in AML cell line, using THP-1 The constructs stabilizing the AML cell line and injected into NSG mice by tail veins, and the AML model was constructed, and chemotherapy drug IDA treatment was administered in the 42nd day after injection. As a result, it was found that there was a significant drug resistance when Mettl3 overexpressed AML mice was treated, while Mettl3 methyltransferase functional mutation, its ability to induce AML chemotherapy resistance ( image 3 . It is suggested that Mettl3 can be used as a treatment target of AML chemotherapy resistance.

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Abstract

The invention particularly relates to application of METTL3 in AML chemotherapy drug resistance, and belongs to the technical field of molecular biology and medicine. The clinical treatment effect of AML is poor, the prognosis condition is poor, and relapse and refractory cases cannot be relieved through conventional therapy. The invention provides a molecular marker for detecting the chemotherapy sensitivity of acute myelogenous leukemia by analyzing the expression difference of METTL3 in bone marrow of a patient relatively sensitive to AML chemotherapy and a patient resistant to AML chemotherapy. Through AML chemotherapy sensitive cell strains, chemotherapy drug-resistant cell strains and animal models, the AML chemotherapy drug-resistant treatment effect of the METTL3 inhibitor is proved. The research of the invention also proves that METTL3 can mediate AML chemotherapy resistance by regulating AML cell homing.

Description

Technical field [0001] The present invention belongs to the field of molecular biology and medical technology, and the application involving Mettl3 in chemotherapy resistance of acute myeloid leukemia (AML), which specifically relates to a molecular marker, kit, kit, and detection method for identifying AML chemotherapy sensitivity. And the application of Mettl3 inhibitors in the preparation of anti-AML chemotherapy drug resistance drugs. Background technique [0002] The information of this background is intended to increase the understanding of the overall background of the present invention without being considered to admit or admit or imply that the information is intended to be a prior art known to those of ordinary skill in the art. [0003] Acute myeloid leukemia, AML is the most common type of acute leukemia, chemotherapy is still the main means of clinical treatment. Currently, only 20-30% of AML patients can reach long-term disease-free survival, dfs, while AML is resis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11A61K45/00A61P35/00
CPCC12Q1/6886C12Q1/6851A61K45/00A61P35/00C12Q2600/158C12Q2600/118C12Q2600/136C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 纪春岩叶静静孙涛李明颖
Owner SHANDONG UNIV QILU HOSPITAL
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