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Application of AK2 gene in preparation of leukemia induced differentiation treatment medicine

A technology for inducing differentiation and treating drugs, applied in the field of medicine and biology, it can solve problems such as unclear relationship, and achieve the effect of reducing the nucleocytoplasmic ratio, increasing the expression, and enhancing the reducing ability.

Pending Publication Date: 2022-07-08
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the relationship between the AK2 gene and the differentiation of leukemia cells is still unclear, and further research is needed

Method used

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  • Application of AK2 gene in preparation of leukemia induced differentiation treatment medicine
  • Application of AK2 gene in preparation of leukemia induced differentiation treatment medicine
  • Application of AK2 gene in preparation of leukemia induced differentiation treatment medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Two AK2 siRNAs targeting different sequences (SEQ ID NO: 3; SEQ ID NO: 4) and negative control nc were introduced into HL60 cells by lipofection (purchased from the Chinese Academy of Sciences Cell Bank), 72 After 1 hour, the cells were harvested, and the cells were lysed with cell lysate to extract proteins, and then Western blot was performed with AK2 antibody (purchased from Proteintech). The results showed that all the above AK2 siRNAs (SEQ ID NO: 3; SEQ ID NO: 4) could effectively inhibit the protein expression of AK2. See the results figure 1 .

Embodiment 2

[0022] Two AK2 siRNAs targeting different sequences (such as SEQ ID NO: 3; SEQ ID NO: 4) and negative control nc were introduced into leukemia cells HL60 by electroporation, and the cells were collected on D3, D5 and D7 days after treatment, and detected. Expression of cell surface differentiation-specific antigens CD11b and CD14. The cells were washed with ice-cold PBS for 3 times, blocked with 3% BSA for 30 min at room temperature, and then incubated with CD11b-PE-labeled monoclonal antibody in the dark for 45 min. After washing with PBS, the cells were detected by flow cytometry and analyzed by CellQuest Pro software. . The results showed that AK2 siRNA could significantly promote the expression of leukemia cell surface specific antigens CD11b and CD14. See the results figure 2 .

Embodiment 3

[0024] Two AK2 siRNAs targeting different sequences (such as SEQ ID NO: 3; SEQ ID NO: 4) and the negative control nc were introduced into leukemia cells HL60 by electroporation, and the cells were collected on D7 days after treatment, and the cells were detected for NBT. Restoration ability. The cells were washed three times with ice PBS and then resuspended with 200 ul PBS, and then 200 ul NBT reaction solution (1 ml PBS containing 2 mg NBT and 2 ug TPA) was added, and the cells were reacted at 37° C. for 40 min in the dark. After that, 1 ml of PBS was added to stop, the supernatant was removed after centrifugation, fixed with methanol, and then applied to a 24-well plate, observed under a microscope and photographed. The results showed that AK2 siRNA could significantly promote the NBT reducing ability of leukemia cells. See the results image 3 .

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Abstract

The invention discloses application of an AK2 gene in preparation of a leukemia induced differentiation treatment medicine. According to the invention, the differentiation of leukemia cells can be directly induced by reducing the expression of AK2 by using an siRNA interference technology. Therefore, it is proposed for the first time that the AK2 gene is a key gene for leukemia induced differentiation treatment, siRNAs molecules of the AK2 can be used as a leukemia induced differentiation agent, a new treatment target is provided for induced differentiation treatment of leukemia, and the AK2 gene plays an important role in treatment of leukemia. The invention provides possibilities for preparing novel drugs for induced differentiation of leukemia, improving the curative effect of leukemia patients and improving drug resistance and related prognosis conditions.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and particularly relates to the application of AK2 (adenylate kinase 2) gene in the preparation of leukemia induction and differentiation therapeutic drugs. Background technique [0002] Acute myeloid leukemia (AML) is a malignant clonal disease characterized by abnormal proliferation of primitive and immature myeloid cells in the bone marrow and peripheral blood. The current standard therapy is anthracycline antibiotics (Anthracycline) combined with cytarabine (Cytarabine) for induction therapy, followed by chemotherapy or stem cell transplantation for consolidation therapy. However, even under this intensive treatment, many patients have extremely poor prognosis and eventually develop relapsed / refractory tumors, and the 5-year survival rate of adult AML patients is only 30%. A hallmark feature of acute myeloid leukemia is the failure of differentiation, in which hematopoietic stem...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/713A61K45/00A61P35/00A61P35/02
CPCA61K31/713A61K45/00A61P35/00A61P35/02
Inventor 应美丹何俏军杨波邵雪晶曹戟许爱笑
Owner ZHEJIANG UNIV
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