Application of miRNA to diagnosis and treatment of acute myelogenous leukemia

An acute myeloid and leukemia technology, applied in the application field of miRNA-1277 in the diagnosis and treatment of acute myeloid leukemia, can solve the problems of inducing cancer and abnormal gene expression, etc.

Active Publication Date: 2015-12-16
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The abnormal expression of miRNA directly leads to the abnormal expression of some genes related to the occurrence of diseases, and induces the occurrence of cancer.

Method used

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  • Application of miRNA to diagnosis and treatment of acute myelogenous leukemia
  • Application of miRNA to diagnosis and treatment of acute myelogenous leukemia
  • Application of miRNA to diagnosis and treatment of acute myelogenous leukemia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Screening of miRNAs associated with acute myeloid leukemia

[0047] 1. Sample acquisition: Collect 10 blood samples from healthy people and blood samples from patients with acute myeloid leukemia. The acquisition of all the above samples was approved by the ethics committee of the organization.

[0048] 2. Extraction of total RNA from samples

[0049] Total RNA was extracted using the BloodRNA extraction kit from U-gene. Specific steps are as follows:

[0050] 1) Add 5 times the volume of 1×XR-I buffer per volume of fresh blood (maximum 1ml), for example: add 5ml of XR-I buffer per 1ml of blood, and vortex to mix;

[0051] 2) Ice-bathed for 15 minutes, quickly mixed twice on a vortex shaker, the solution became clear, indicating that the red blood cells had been lysed. If the hematocrit or ECR of individual samples increases, extend the ice bath time to 20 minutes;

[0052] 3) Centrifuge at 450g for 10min at 4°C to precipitate white blood cells, and compl...

Embodiment 2

[0078] Example 2 QPCR verification of differentially expressed miRNA-1277

[0079] 1. According to the detection results of the miRNA chip, select miRNA-1277 for large sample QPCR verification. According to the sample collection method in Example 1, 80 blood samples from patients with acute myeloid leukemia and 80 blood samples from healthy people were selected.

[0080] 2. The RNA extraction process is the same as in Example 1.

[0081] 3. Reverse transcription:

[0082]1) Prepare the reaction system according to Table 1

[0083] Table 1 reaction system

[0084] RNA template

1μg

10× buffer

2μl

dATP (10mM)

2μl

polyA polymerase

0.5μl

RNase inhibitor

0.5μl

wxya 2 o

Make up to 20μl

[0085] Incubate at 37°C for 1h.

[0086] 2) Add 1 μl of 0.5 μg / μl Oligo(dT) specific RT primer to the reaction tube, and incubate at 70° C. for 5 minutes.

[0087] 3) Immediately incubate on ice for 2 minutes to break ...

Embodiment 3

[0104] Example 3 Expression of miRNA-1277 in acute myeloid leukemia cell lines

[0105] 1. Cell culture

[0106] Acute myeloid leukemia cell lines U937, KG-1, HL-60 and bone marrow mononuclear cells isolated from AML patients and healthy people were routinely cultured in RPMI1640 medium containing 10% fetal bovine serum, placed at 37°C, 5%CO 2 in the incubator.

[0107] 2. QPCR

[0108] 2.1 Extraction of total cellular RNA: Extraction of total cellular RNA was performed using the RNA extraction kit from QIAGEN, following the instructions in the instructions.

[0109] 2.2 QPCR: the steps are the same as in Example 2.

[0110] 3. Results

[0111] Such as figure 2 As shown, compared with the bone marrow mononuclear cells of healthy people, the expression of miRNA-1277 in the bone marrow mononuclear cells of acute myeloid leukemia cell lines U937, KG-1, HL-60 and AML patients was significantly decreased (P<0.05).

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Abstract

The invention discloses application of miRNA to diagnosis and treatment of acute myelogenous leukemia. The miRNA is miRNA-1277. Whether a patient has the acute myelogenous leukemia or not can be diagnosed through detecting the expression level of the miRNA-1277. Experiments prove that the expression level of the miRNA-1277 in the patient with the acute myelogenous leukemia is obviously lower than that of healthy people. On the basis, the miRNA-1277 can also be used for preparing medicine for treating the acute myelogenous leukemia. The sensitivity and the specificity of the diagnosis of the acute myelogenous leukemia are greatly improved; meanwhile, a new target point is provided for gene treatment of the acute myelogenous leukemia.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of miRNA in the diagnosis and treatment of acute myeloid leukemia, in particular to the application of miRNA-1277 in the diagnosis and treatment of acute myeloid leukemia. Background technique [0002] Acute leukemia is a clonal malignant disease originating from hematopoietic stem cells. The abnormal proliferation of leukemia cells in the bone marrow leads to the suppression of normal hematopoiesis. The clinical manifestations are anemia, bleeding, infection, etc. Acute myeloid leukemia (AML) is the most common adult acute leukemia. In recent years, the incidence of AML has gradually increased. Although the complete remission rate of patients with existing treatment methods can reach 50% to 70%, the mortality rate of the disease is still high. Higher, the implementation of individualized stratified treatment for different patients has become a concern. Comprehensive rese...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68A61K48/00A61K31/7105A61P35/00
CPCC12N15/113C12N2310/141C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 边洋杨承刚
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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