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Application of miR-6812-5p in preparing drugs for treating leukemia

A technology of drugs and recombinant viruses, applied in application, gene therapy, drug combination, etc., can solve the problem of lack of effective drugs for the treatment of metastatic leukemia

Inactive Publication Date: 2017-01-04
BEIJING BELIFE BIO MEDICAL TECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Clinically, surgery or radiotherapy is effective for localized leukemia, and there is still a lack of effective drugs for the treatment of metastatic leukemia

Method used

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  • Application of miR-6812-5p in preparing drugs for treating leukemia
  • Application of miR-6812-5p in preparing drugs for treating leukemia
  • Application of miR-6812-5p in preparing drugs for treating leukemia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1. Obtainment of Metastatic Leukemic Cell Lines Stable and Highly Expressing miR-6812-5p

[0031] 1. Acquisition of leukemia cell lines

[0032] For the colorectal cancer cell line Daudi (ATCC), the culture medium is 1640 medium (Invitrogen, 31800-022) containing 10% fetal bovine serum (Hyclone) and 0.2% penicillin and streptomycin (Invitrogen).

[0033] A. Cell recovery

[0034] (1) Take out the cryopreservation tube of Daudi (ATCC), immediately put it in a water bath at 37°C-40°C, and shake it quickly until it melts completely.

[0035] (2) Transfer the cell suspension into 10 mL of cold PBS, put it into a 15 mL centrifuge tube, centrifuge the cells with a horizontal centrifuge (1000 rpm, 8 minutes) and discard the supernatant.

[0036] (3) Suspend the cells with an appropriate amount of fresh medium, transfer the cell suspension into a culture bottle, and culture in a carbon dioxide incubator at 37°C.

[0037] B. Cell culture

[0038] Put the leukemia cel...

Embodiment 2

[0062] Example 2. Functional study of stable and high expression of miR-6812-5p in Daudi-high cell line

[0063] 1. Cell growth curve

[0064] (1) 24 hours after transfection, the Daudi-high obtained in Example 1 in a good growth state was taken and digested by a general passage method to prepare a cell suspension. Counting, accurately planting cells in a 96-well plate, the total number of cells per well is required to be consistent (2000 / well), the amount of culture medium added should also be consistent, and a blank control (that is, the same volume of medium) is set, each well The final volume is 100 μL. Set at 37°C, 5% CO 2 Incubate for 24 hours in the incubator.

[0065] (2) Add 10 ul of CCK-8 solution (purchased from Japan Dojin Chemical Research Institute, article number: CK04) to each well. The amount of CCK-8 solution can be increased or decreased according to specific conditions.

[0066] (3) Normal culture conditions (37°C, 5% CO 2 incubator) for 4 hours (the ...

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Abstract

The invention provides application of any one of substances in (1) to (4) in preparing a product for promoting migration, spreading and / or invasion of cancer cells: (1) miR-6812-5p; (2) a recombinant carrier containing an encoded gene of miR-6812-5p; (3) a recombinant virus containing the encoded gene of miR-6812-5p; and (4) a recombinant virus carrier containing the encoded gene of miR-6812-5p. An experiment proves that the overexpression of the miR-6812-5p cannot influence the growth of Daudi cells, but can remarkably enhance the invasion and migration capacity of leukemia cells and can play an important role in the transfer process of the leukemia; and therefore, the miR-6812-5p is associated with the transfer of the leukemia.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of microRNA in the preparation of medicines for treating leukemia. Background technique [0002] microRNA (miR) is a single-stranded small molecule RNA with a size of about 21-23 bases. It guides the silencing complex (RISC) to degrade mRNA or inhibit the translation of mRNA by base pairing with target gene mRNA, usually after transcription Horizontal regulation of protein expression (recently found that microRNA can also regulate gene expression at the transcriptional level). With the deepening of microRNA research, more and more evidences show that microRNA molecules play the role of proto-oncogene or tumor suppressor gene in all aspects of tumorigenesis and development. [0003] Leukemia is a malignant clonal disease of hematopoietic stem cells. Clonal leukemia cells proliferate and accumulate in bone marrow and other hematopoietic tissues due to mechanisms such ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/867C12N5/10A61K48/00A61K31/7105A61P35/02C12Q1/02
CPCC12N15/1135A61K31/7105C12N5/0693C12N2310/141G01N33/5011
Inventor 崔健姜薇
Owner BEIJING BELIFE BIO MEDICAL TECH LTD