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Application of three microRNAs in diagnosis and treatment of human acute myeloid leukemia

A technology for acute myeloid and leukemia, applied in gene therapy, microbial determination/examination, biochemical equipment and methods, etc., can solve problems such as no research report yet

Inactive Publication Date: 2010-11-17
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0013] Although there have been research reports on miRNAs and AML in the world, the specific miRNAs screened out do not involve miR-29a, miR-142-3, and miR-451, so there is no research report on the relationship between these three miRNAs and AML.

Method used

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  • Application of three microRNAs in diagnosis and treatment of human acute myeloid leukemia
  • Application of three microRNAs in diagnosis and treatment of human acute myeloid leukemia
  • Application of three microRNAs in diagnosis and treatment of human acute myeloid leukemia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: Experimental object

[0038] All AML patients were selected from the visiting patients by the Department of Hematology, Affiliated Hospital of Inner Mongolia Medical College. The diagnosis of AML was based on conventional methods, and the morphological diagnosis and typing of each case were in line with the requirements of the "Standards for the Diagnosis and Curative Effect of Hematological Diseases". The age and gender-matched normal controls were selected from volunteer blood donors. In addition to normal hemogram indicators to exclude blood diseases, other important diseases were also excluded. All followed the principle of informed consent. The research plan was approved by the ethics review committees of both parties (Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and the Affiliated Hospital of Inner Mongolia Medical University).

Embodiment 2

[0039] Example 2: Detection of Abnormally Expressed miRNAs in AML Patients Using miRNA Chip Analysis

[0040] In order to compare the expression differences of miRNAs in AML patients and normal controls, we first used the miRNA microarray analysis method. The samples used for microarray analysis were 2 cases of newly diagnosed AML (M5 type) cases and 2 cases of normal controls matched in gender and age. Isolate the mononuclear cells of its peripheral blood (the separation method is described in detail in the next paragraph), and entrust Shanghai Kangcheng Bioengineering Co., Ltd. to perform the miRNA chip experiment operation (using Exiqon, LNA TM microRNA chip) and provide complete experimental and analysis reports.

[0041] Peripheral blood mononuclear cells (MNC) were isolated by density gradient centrifugation. The collected fresh peripheral blood was diluted with 1×phosphate-buffered saline (PBS) containing 2mmol / L EDTA according to the volume of 1:4, and the pipette wa...

Embodiment 3

[0042] Example 3: Northern hybridization verification of chip results

[0043] In order to eliminate the false positive results in chip analysis, we verified some of the results by Northern hybridization. Total RNA is first extracted from a mononuclear cell sample. at approximately every 1 x 10 7 1 ml Trizol (purchased from Invitrogen) was added to each mononuclear cell, and the cells were immediately repeatedly pipetted and left at room temperature for 5 min to lyse. Add 0.2 ml of chloroform to the lysis pipette, vibrate the centrifuge tube on a vortex shaker for 15 s, let it stand at room temperature for 2-3 min, and then centrifuge at 12,000×g for 15 min at 4°C. Transfer the upper aqueous phase to a new centrifuge tube, add 0.5ml of isopropanol to mix, and place at room temperature for 10 minutes. Centrifuge at 12,000×g for 10 min at 4°C and discard the supernatant. Rinse the precipitate once with 75% ethanol, centrifuge at 7500×g for 5 min, discard the supernatant, and...

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Abstract

The invention relates to an application of three miRNAs, i.e. miR-451, miR-29a and miR-142-3p in diagnosis and treatment of acute myeloid leukemia (AML).

Description

technical field [0001] The present invention relates to the function and application of three microRNAs, more specifically, the present invention relates to miR-451, miR-29a and miR-142 whose expression is significantly reduced in peripheral blood mononuclear cells of human acute myeloid leukemia (AML) patients Function of -3p and its application in diagnosis, prognosis and treatment of AML. Background technique [0002] miRNA is a class of non-coding small molecule RNA newly discovered in recent years, and its length is usually 19-25 nucleotides (nt). In the nucleus, the genes encoding miRNAs are first transcribed into primary transcripts under the action of RNA polymerase II, and then undergo a series of cleavage and transport to the cytoplasm to produce mature miRNA molecules. These miRNA molecules inhibit translation or degrade target gene mRNA molecules through complementary pairing (Voinnet O, Cell. 136:669-87, 2009). Since the first miRNA molecule was discovered in ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68A61K48/00A61P35/02
Inventor 张俊武王芳杨桂花王小爽肖镇高大黄彬涛王静
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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