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Application of cdk2 gene in preparation of medicine for inducing differentiation therapy of leukemia

A technique for inducing differentiation and treating drugs, which is applied in the field of medicine and biology, can solve problems such as unclear relationship between CDK2 genes, and achieve the effect of improving drug resistance and improving curative effect

Active Publication Date: 2016-04-20
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the relationship between the CDK2 gene and the differentiation of leukemia cells is still unclear and needs further study

Method used

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  • Application of cdk2 gene in preparation of medicine for inducing differentiation therapy of leukemia
  • Application of cdk2 gene in preparation of medicine for inducing differentiation therapy of leukemia
  • Application of cdk2 gene in preparation of medicine for inducing differentiation therapy of leukemia

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 13 CDK2 siRNAs (SEQIDNO:2-SEQIDNO:14) targeting different sequences and the negative control nc were introduced into COS7 cells (purchased from the Cell Bank of the Chinese Academy of Sciences) by lipofection, and the cells were harvested 72 hours later , and the cells were lysed with cell lysate to extract proteins, and then Western blot was performed with CDK2 antibody (purchased from Santa Cruz). It was found that all the above CDK2 siRNAs can effectively inhibit the protein expression of CDK2. See results figure 1 .

Embodiment 2

[0036] Three CDK2 siRNA targeting different sequences with different silencing efficiencies (such as SEQIDNO:2; SEQIDNO:4; SEQIDNO:9, etc.) and negative control nc were introduced into leukemia NB4 cells (Dr. Lingtao Wu, University of Southern California, USA) by electroporation, After the 1st, 2nd, 3rd, and 5th days of treatment, blood cells were counted using a hemocytometer. The results show that CDK2siRNA can significantly inhibit the proliferation of leukemia NB4 cells, see figure 2 .

Embodiment 3

[0038] With different concentrations (2.5′10 -6 -1'10 -5M) Leukemic cell lines (NB4, U937) were treated with CDK2 inhibitors (SU-9516, CVT-313). After 72 hours, 5 mg / ml MTT solution was added and incubated for another 4 hours. Then the liquid was discarded and DMSO was added to dissolve After the formazan was completely dissolved, the absorbance at 570 nm was measured with a microplate reader. It was found that CDK2 inhibitors (SU-9516, CVT-313) acted on leukemia cells (NB4, U937) (U937 cells were purchased from the Cell Bank of the Chinese Academy of Sciences) and could significantly inhibit cell proliferation in a concentration-dependent manner. see image 3 .

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Abstract

The invention aims to provide a CDK2 gene, and particularly relates to the application of CDK2siRNA and a CDK2 inhibitor to the preparation of leukemia induced differentiation therapeutic drug. The drug takes a CDK2 gene as a target spot. The CDK2 gene provided by the invention can restrain the protein expression level or activity of CDK2, and further obviously induces the differentiation of leukemia cells of people, in particular, cell proliferation is restrained, the expression of specific antigen CD11b differentiated on the cell surface is remarkably increased, and cell amount of a segmented nucleus is increased; based on the application, a new therapeutic target and new effective drugs are provided for the induced differentiation leukemia treatment.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and in particular relates to the application of CDK2 gene in the preparation of medicines for inducing differentiation of leukemia. Background technique [0002] Leukemia is a kind of clonal malignant disease with abnormal hematopoietic stem cells. Its important sign is severe differentiation disorder, that is, leukemia cells lose the ability to further differentiate and mature and stagnate at different stages of cell development. Therefore, as early as the 1970s, the scientist Sachs proposed the induction of differentiation therapy. At present, the use of all-trans retinoic acid (all-transretioindacid, ATRA) in the treatment of acute promyelocytic leukemia is the most successful example of differentiation therapy strategy in clinical practice. However, there are still major limitations in the development of drugs for induction of differentiation therapy, including a small number of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K38/00A61K45/00A61K31/4178A61P35/02
Inventor 何俏军杨波应美丹邵雪晶曹戟罗沛华
Owner ZHEJIANG UNIV
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