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Specific miRNA related to M5 acute myeloid leukemia and its application

An acute myeloid and leukemia technology, applied in specific miRNA and its application fields, can solve the problems of single detection method, affecting the accuracy of diagnosis, delaying the disease, etc., and achieve the effect of broad application prospects and good clinical application value

Active Publication Date: 2018-06-26
BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, M5 type AML (M5 acute monocytic leukemia) is mainly detected through morphological aspects such as blood picture / bone marrow picture and cytochemical staining. Compared with other types of AML, the detection method is simpler, which greatly affects the accuracy of diagnosis. rate, which brings great uncertainty to the clinical treatment and medication guidance of patients with this type of AML, and delays the treatment of patients.

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  • Specific miRNA related to M5 acute myeloid leukemia and its application
  • Specific miRNA related to M5 acute myeloid leukemia and its application
  • Specific miRNA related to M5 acute myeloid leukemia and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Screening of highly expressed miRNAs in M5 acute myeloid leukemia cell lines

[0039] By analyzing and comparing M5 leukemia cell lines (THP-1, THP-1 derived from patients with acute monocytic leukemia) and non-M5 myeloid leukemia cell lines (K562, K562 derived from patients with chronic myeloid leukemia) Through-put miRNA-seq data (NCBI accession number: GSE48059) screened 16 highly expressed miRNAs in M5 AML cell lines.

[0040] The above-mentioned 16 miRNA-specific primer pairs were designed, and the expression levels of each miRNA were verified by ABI 7500 fluorescent quantitative PCR instrument, using the RNA tailed reverse transcription products of M5 type THP-1 cell line and non-M5 type K562 cell line as templates. The following table lists 16 miRNAs and their specific amplification primers.

[0041] Table 1

[0042]

[0043]

[0044] Comparing M5 type acute myeloid leukemia cell line (THP-1) and non-M5 type chronic myeloid leukemia cell line (K...

Embodiment 2

[0048] Example 2 Establishment of RT-PCR detection method for M5 acute myeloid leukemia

[0049] 1. Sample collection: 12 bone marrow samples from M5 AML patients and 1 normal bone marrow sample.

[0050] 2. Sample processing: extract total RNA according to the standard operation of Trizol reagent.

[0051] 3. Sample detection: Take 2 μg of total RNA.

[0052] (1) RNA tailing Mix the ingredients in Table 2 in a 0.2mL PCR tube, then bathe in water at 37°C for 1.5 hours.

[0053] Table 3 RNA tailing reaction system.

[0054] Reagent

volume

RNA (DNA free)

2μg

10×Poly(A)Polymerase Reaction Buffer

1μL

ATP (10mM)

1μL

E.coli Poly(A) Polymerase (5U / μl)

0.2uL

Add decalculated water to make up to

20 μL

[0055] (2) Add reverse transcription primers: add 0.5 μl (1 μg / μl) reverse transcription primers, place in a 65-degree water bath for 10 minutes, then place on ice for at least 5 minutes, and keep on ice until R...

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Abstract

The invention provides specificity miRNA related to M5 type acute myelogenous leukemia and application thereof. The new-generation high-throughput sequencing technology is utilized and combined with a systematic bioinformatics analysis method, screening is carried out to obtain the specificity miRNA related to M5 type acute myelogenous leukemia, and the RNA sequence of the specificity miRNA is shown as SEQ ID NO.1-3. The invention further provides a primer for detecting the specificity miRNA and application thereof to preparation of a diagnostic kit for detecting M5 type acute myelogenous leukemia. The specificity miRNA can be applied to clinical diagnosis of leukemia and guidance. The specificity primer for detecting the miRNA can be used for preparing a diagnostic reagent or the diagnostic kit for detecting M5 type acute myelogenous leukemia, drug use guidance can be provided for treating acute myelogenous leukemia, and the specificity miRNA has high application value and wide application prospects.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a specific miRNA related to M5 acute myeloid leukemia (AML) and its application. Background technique [0002] Leukemia is a heterogeneous disease characterized by the abnormal proliferation of blood precursor cells of myeloid or lymphoid origin. Leukemia generally has a poor prognosis and a high mortality rate, so it has attracted the attention of the academic community. Leukemia is a heterogeneous disease characterized by abnormal proliferation of blood precursor cells of myeloid or lymphoid origin, mainly including acute lymphoid leukemia (ALL), acute myeloid leukemia (AML) and chronic lymphoid leukemia (CLL) and chronic myeloid leukemia (CML) etc. (Lightfoot, 2005). In my country, acute leukemia (AL) is more common than chronic leukemia (CL) (about 5.5:1). Acute myeloid leukemia (AML) is more common in young adults, and the incidence of acute lymphoblastic leuke...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12Q1/6886C12N15/11
CPCC12N15/113C12N2310/141C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 方向东张昭军刘金立陈晓娟刘淑阁熊倩王万恒
Owner BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION