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Application of ZDHHC21 genes in preparation of leukemia induced differentiation therapy drugs

A technology for inducing differentiation and treating drugs, which is applied in the field of medicine and biology, can solve the problems of ZDHHC21 gene and leukemia cell differentiation that have not been reported in the literature, and achieve the effect of improving drug resistance and improving curative effect

Active Publication Date: 2019-05-31
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These studies all suggest that ZDHHC protein plays an important regulatory role in the initiation and development of tumors, but the relationship between ZDHHC21 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 ZDHHC21 genes in preparation of leukemia induced differentiation therapy drugs
  • Application of ZDHHC21 genes in preparation of leukemia induced differentiation therapy drugs
  • Application of ZDHHC21 genes in preparation of leukemia induced differentiation therapy drugs

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Experimental program
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Effect test

Embodiment 1

[0023] Three ZDHHC21 siRNAs (SEQ ID NO:3-SEQ ID NO:5) targeting different sequences and the negative control nc were introduced into HL60 cells (purchased from the Chinese Academy of Sciences Cell Bank) by lipofection, 72 The cells were harvested after 1 hour, and the cells were lysed with Trizol lysate to extract mRNA, and then the mRNA level was detected by fluorescent quantitative RT-PCR. It was found that all the ZDHHC21 siRNAs above could effectively inhibit the mRNA expression of ZDHHC21. See results figure 1 .

Embodiment 2

[0025] Two ZDHHC21 siRNA targeting different sequences (such as SEQ ID NO:4; SEQ ID NO:5) and negative control nc were introduced into leukemia HL60 and NB4 cells (Dr. Lingtao Wu, University of Southern California) by electroporation. After D3, D5 and D7, the blood cells were counted with a hemocytometer. The results showed that ZDHHC21siRNA could significantly inhibit the proliferation of leukemia HL60 and NB4 cells. See results figure 2 .

Embodiment 3

[0027] Two ZDHHC21 siRNA targeting different sequences (such as SEQ ID NO:4; SEQ ID NO:5) and negative control nc were introduced into leukemia HL60 and NB4 cells by electroporation, and the cells were collected on D7 days after treatment to detect the cell surface Expression of the differentiation-specific antigen CD11b. Cells were rinsed with ice PBS for 3 times, blocked with 3% BSA for 30 min at room temperature, then incubated with CD11b-PE-labeled monoclonal antibody in the dark for 45 min, washed with PBS, detected by flow cytometry, and analyzed by CellQuestPro software. The results showed that ZDHHC21siRNA can significantly promote the expression of specific antigen CD11b on the surface of leukemia cells. See results image 3 .

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Abstract

The invention provides application of ZDHHC21 genes in preparation of leukemia induced differentiation therapy drugs, and application of polypeptides targeting the DHHC21 genes, small molecule inhibitors and interfering siRNAs in preparation of leukemia induced differentiation agents. Research shows that as the ZDHHC21 amount is reduced by the RNA interfering technology, the proliferation of leukemia HL60 and NB4 cells is significantly inhibited, the expression of cell surface differentiation-specific antigens CD11b is significantly increased, the reduction ability of NBT of the cells is enhanced, and the ratio of nucleus to cytoplasm in the cells is reduced, the shapes of cell nuclei are changed into U shapes or half-moon shapes. Down-regulation of ZDHHC21 can effectively induce the differentiation of the human leukemia cells, a new direction is provided for the development of treatment drugs for leukemia, a new therapeutic target is provided for induction differentiation therapy of leukemia, and a new research field is opened up for the treatment of leukemia, the improvement of the treatment effect and the improvement of drug resistance.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and specifically relates to the application of a ZDHHC21 gene in the preparation of medicines for inducing differentiation of leukemia. Background technique [0002] Leukemias are clonal diseases of myeloid progenitor cells characterized by clinical and biological heterogeneity. After decades of research, although researchers have made great progress in the understanding of the pathology and genomics of acute myeloid leukemia, the treatment of leukemia is still the focus of today's tumor therapy because of its relapse and refractory. One of the toughest challenges. Although scientists have proposed induction therapy for the important characteristics of leukemia differentiation disorder, and all-trans retinoic acid has also achieved great success in APL subtype leukemia, but the current clinical induction therapy is only limited to APL subtype, and There are very few drugs to choose...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K31/713A61K45/00A61K38/17A61K31/7088A61P35/02
Inventor 应美丹邵雪晶何俏军杨波曹戟朱虹
Owner ZHEJIANG UNIV
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