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Application of yod1 gene in preparation of drugs for inducing differentiation therapy of leukemia

A technology for inducing differentiation and treating drugs, which is applied in the field of medicine and biology, can solve the problems of YOD1 gene and leukemia cell differentiation that have not been reported in the literature, and achieve the effect of enhancing NBT reducing ability and reducing the ratio of nucleus to cytoplasm

Active Publication Date: 2021-06-01
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These studies all suggest that YOD1 protein regulates the ubiquitination level of related proteins to participate in important life activities of cells, but the relationship between YOD1 gene and leukemia cell differentiation has not been reported in the literature, and further research is needed

Method used

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  • Application of yod1 gene in preparation of drugs for inducing differentiation therapy of leukemia
  • Application of yod1 gene in preparation of drugs for inducing differentiation therapy of leukemia
  • Application of yod1 gene in preparation of drugs for inducing differentiation therapy of leukemia

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Experimental program
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Effect test

Embodiment 1

[0026] Seven YOD1 siRNAs targeting different sequences (SEQ ID NO:3-SEQ ID NO:9) and the negative control NC were introduced into NB4 cells by lipofection (Dr. Lingtao Wu, University of Southern California, USA). After 72 hours, the cells were harvested, and the cells were lysed with the cell lysate to extract proteins, and then Western blot was performed with YOD1 antibody (purchased from Proteintech). As a result, it was found that all the above YOD1 siRNAs (SEQ ID NO:3-SEQ ID NO:9) could effectively inhibit the protein expression of YOD1. See results figure 1 .

Embodiment 2

[0028] Two YOD1 siRNAs targeting different sequences with different silencing efficiencies (SEQ ID NO:3,SEQ ID NO:4) and the negative control NC were introduced into leukemia NB4, U937 and HL60 cells by lipofection (purchased from the Cell Bank of the Chinese Academy of Sciences). After treatment D1, D3 and D5, counts were performed using a hemocytometer. The results showed that YOD1 siRNA (SEQ ID NO:3, SEQ ID NO:4) could significantly inhibit the proliferation of leukemia NB4, U937 and HL60 cells. See results figure 2 .

Embodiment 3

[0030] YOD1 siRNA (SEQ ID NO:3, SEQ ID NO:4) targeting different sequences with different silencing efficiencies and the negative control NC were introduced into NB4 cells by lipofection. The cells were collected after treatment on D3 and D5, and the expression of the differentiation-specific antigen CD11b on the cell surface was detected. Cells were rinsed with ice PBS for 3 times and then incubated with 1% BSA and CD11b-PE-labeled monoclonal antibody in the dark for 60 min. After washing with PBS, the cells were detected by flow cytometry and analyzed by CellQuest Pro software. As a result, it was found that YOD1 siRNA (SEQ ID NO:3, SEQ ID NO:4) can significantly promote the expression of the specific antigen CD11b on the surface of leukemia cells. See results image 3 .

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Abstract

The present invention provides an application of YOD1 gene in the preparation of a leukemia induction and differentiation therapeutic drug. The YOD1 gene includes a polypeptide targeting the YOD1 gene, a small molecule inhibitor targeting the YOD1 protein, and interfering siRNAs targeting the YOD1 gene. The present invention utilizes siRNA interference technology to induce differentiation of leukemia NB4 cells by reducing the expression of YOD1. Down-regulation of YOD1 can effectively induce the differentiation of human leukemia cells, which provides a new direction for the development of current therapeutic drugs for leukemia. It opens up a new research field, and provides the possibility to prepare new drugs for inducing differentiation of leukemia, improve the curative effect of leukemia patients, and improve drug resistance and related prognosis. The invention provides a new therapeutic target for inducing differentiation and treating leukemia.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and specifically relates to the application of a YOD1 gene in the preparation of medicines for inducing differentiation of leukemia. Background technique [0002] Leukemia is a class of malignant clonal diseases of hematopoietic stem cells characterized by clinical and biological heterogeneity and severe differentiation disorders. After decades of research, researchers have made some progress in understanding the pathology and genomics of leukemia, and aiming at the important characteristics of leukemia differentiation disorder, scientists have proposed induction differentiation therapy for the treatment of leukemia. Currently, differentiation therapy using all-trans retinoic acid has achieved great success in the APL subtype of leukemia. However, the application of clinical induction differentiation therapy is currently limited to APL subtypes, and there are very few drugs to choos...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61K38/16A61K31/7088A61P35/02
CPCA61K31/7088A61K38/16A61K45/00A61P35/02
Inventor 应美丹何俏军杨波邵雪晶陈颖倩曹戟朱虹
Owner ZHEJIANG UNIV
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