Application of miR-638 in resisting acute myelogenous leukemia

A technology of 1.mir-638 and 2.mir-638, applied in the field of molecular biology, can solve problems such as cell cycle abnormality, disease recurrence and mortality

Inactive Publication Date: 2014-12-17
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

(2) Abnormal expression of miRNA leads to abnormal cell cycle
The methylation of miRNA leads to the down-regulation of miR-124 expression in ALL patients, which in turn can dow

Method used

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  • Application of miR-638 in resisting acute myelogenous leukemia
  • Application of miR-638 in resisting acute myelogenous leukemia
  • Application of miR-638 in resisting acute myelogenous leukemia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 Investigation of the expression of miR-638 in blood cells from different sources

[0068] In order to explore whether miR-638 is involved in the normal hematopoietic process and the development of leukemia, this example isolated and detected the background expression level of miR-638 in the myeloid, lymphoid and hematopoietic pluripotent stem cell populations in the peripheral blood of healthy people ; and the expression levels in peripheral blood mononuclear cells of healthy controls, various leukemia cell lines, and peripheral blood blasts of AML patients.

[0069] Peripheral blood and umbilical cord blood of AML patients were obtained from Zhongnan Hospital of Wuhan University, Tongji Hospital Affiliated to Huazhong University of Science and Technology, and Jiangsu Provincial Hospital of Traditional Chinese Medicine. Each sample obtained the informed consent of the patient and the approval of the corresponding medical ethics committee. Leukemic cell lines i...

Embodiment 2

[0076] Example 2 Analysis of expression changes of miR-638 in the process of induction and differentiation of leukemia cells and peripheral blood blast cells of AML patients

[0077] 1. Analysis of expression changes of miR-638 during induction and differentiation of leukemia cells

[0078] In order to further verify the correlation between miR638 and the occurrence and development of leukemia, this example uses the in vitro differentiation model of leukemia cells to analyze the relationship between the expression of miR-638 and the differentiation of leukemia cells. HL-60 and NB4 cells can differentiate into granulocytes under the treatment of all-trans retinoic acid (ATRA); THP-1 and HL-60 cells can differentiate into mononuclear cells under the induction of phorbol ester (PMA). Cell Differentiation. The morphological changes of leukemia cells during differentiation were observed by Wright's Giemsa staining. image 3 It shows the cell morphology changes when THP-1 and HL-6...

Embodiment 3

[0083] Example 3 The expression changes of miR-638 in leukemia cells significantly affect the cell differentiation process

[0084] In order to study the effect of miR-638 on the differentiation induction of myeloid leukemia cell lines in vitro, this example artificially regulates the cells (THP-1 and HL-60) the expression level of miR-638. miRNA mimics (mimics) and inhibitors (inhibitors) and their respective controls were purchased from Ribio Biotechnology Co., Ltd., Guangzhou, Guangdong, China. The mimic sequence was completely consistent with the mature miR-638 sequence, and the inhibitor sequence was identical to the mature miR-638 sequence. The sequence of miR-638 is perfectly complementary. By using IfectRNA Plus transfection reagent (product of PharmInova Company) and BLOCK-iT fluorescent molecule (FITC) labeled oligonucleotide (product of Invitrogen Company), the transfection efficiency of these oligonucleotides has reached above 90%.

[0085] By semi-quantitative (...

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Abstract

The invention discloses application of miR-638 in resisting myelogenous leukemia, and belongs to the field of molecular biology. Experiments indicate that in the processes of normal hematopoietic cell differentiation and development as well as acute myelogenous leukemia occurrence and development, the expression level of miR-638 has remarkable changes; in leukemic cell lines and primary acute myelogenous leukemia cells, the expression level of the miR-638 is up-regulated or down-regulated, and the cell differentiation process can be remarkably promoted or inhibited. Researching results indicate that the aim of treating acute myelogenous leukemia can be achieved by up-regulating the expression of the miR-638. Medicines for resisting acute myelogenous leukemia can be prepared from the miR-638, or the analog or expression vector of the miR-638 based on the functions of the miR-638. Important foundation is provided to development of novel acute myelogenous leukemia resisting medicines or diagnostic reagents.

Description

technical field [0001] The invention belongs to the field of molecular biology and relates to the application of a primate-specific non-coding microRNA—hsa-miR-638 (hereinafter referred to as miR-638) in anti-acute myeloid leukemia. Background technique [0002] MicroRNAs (microRNAs, miRNAs) are a class of single-stranded small RNA molecules with a length of about 19-25 nucleotides (nt), which mainly regulate post-transcriptional gene expression by inhibiting protein translation or disrupting the stability of target gene transcripts. In 1993, the first miRNA—lin-4 was discovered in C. elegans by two research groups respectively, and it was officially named microRNA in 2001. A large number of subsequent studies have shown that the biological function of miRNAs involves almost all aspects of animal and plant life activities, including embryonic development and tumorigenesis. Embryonic development is the only way for animals and plants to go from fertilized eggs to adults; tum...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K45/00C12Q1/68A61P35/02
Inventor 李文鑫郭明雄黄赞林义
Owner WUHAN UNIV
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