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Application of nucleoside cytokinin in human acute myeloid leukemia M4 cells

A cytokinin and acute leukemia technology, applied in the application field of nucleoside cytokinin in human acute myeloid leukemia M4 cells, can solve the problems of no research on promoting differentiation, unstable curative effect, and easy relapse of patients , to achieve significant in vitro proliferation inhibitory activity

Inactive Publication Date: 2018-09-21
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the effects of natural CK and their analogues on animal cells have been reported many times, there is no systematic study on the differentiation-promoting effects of some nucleoside forms of cytokinins
[0003] Acute myeloid leukemia (AML) is a clonal hematological malignancy that develops into myeloid cells and is characterized by uncontrolled proliferation and blockage of normal hematopoietic cell differentiation. AML can be divided into subtypes such as M0-M7 , all-trans retinoic acid (ATRA) can cure a unique AML subtype---acute promyelocytic leukemia (APL) by inducing differentiation, however, ATRA-mediated differentiation therapy is not applicable to other types of AML and ATRA Induction therapy still has problems such as unstable curative effect and easy relapse in some patients

Method used

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  • Application of nucleoside cytokinin in human acute myeloid leukemia M4 cells
  • Application of nucleoside cytokinin in human acute myeloid leukemia M4 cells
  • Application of nucleoside cytokinin in human acute myeloid leukemia M4 cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Effects of oTR, KR, BAR and iPR on the expression of CD11b in HL-60 cells: The resuscitated human M4 leukemia cell THP-1 was cultured in a medium containing 10% FBS inactivated fetal bovine serum, 1% penicillin / 1% streptomycin (double antibody) sterile DMEM medium, placed in a suspension cell bottle, at 37 ° C, 5% CO 2 and cultured in an incubator under saturated humidity, and the cells grow to about 70%-80% for one passage. Continue culturing to obtain active human leukemia cells, and collect the cells. 5×10 that will be in logarithmic growth phase 4 Each cell was seeded in a 24-well plate containing 1ml of DMEM culture medium in each well. According to the experimental requirements, corresponding concentrations of oTR, KR, BAR, and iPR were added to each group, and they were incubated in an incubator for 36 hours. Wash three times with FBS and PBS. Cells were incubated with blocking antibody for 15 min at room temperature, then washed after incubation with anti-hum...

Embodiment 2

[0023] Morphological changes of HL-60 cells induced by oTR, KR, BAR and iPR: Actively growing THP-1 cells were treated with 10 μM oTR, KR, BAR and iPR for 36 h, and the cells were collected and washed three times with PBS containing 0.2% FBS. Slides were fixed with methanol and stained with Wright-Giemsa staining solution for 20 minutes, rinsed with distilled water, air-dried and visualized using a microscope. The result is as figure 2 as shown, figure 2 Visible, untreated THP-1 cells as highly malignant cells, round, large and round nuclei and sparse cytoplasm. Treatment with oTR, KR, BAR, iPR decreased the cytoplasmic ratio of nuclei and altered the horseshoe morphology of nuclei. The role of oTR, KR, BAR, iPR in inducing differentiation was further confirmed by morphological analysis using Wright Giemsa staining.

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Abstract

The invention provides an application of nucleoside cytokinin in human acute myeloid leukemia M4 type cells, and belongs to the technical fields of biochemistry and molecular biology. Data in the invention is established on the basis of oTR, KR, BAR and iPR having very significant in-vitro propagation inhibition activity to sixty tumor cell lines in nine human tumor systems and having strongest drug susceptibility to leukemia cell strains.

Description

technical field [0001] The present invention relates to nucleoside cytokinin ortho-Topolin Riboside (oTR), Kinetinriboside (KR), N 6 -Anti-tumor effects of Benzyladenosine (BAR) and N6-Isopentenyladenosine (iPR) on human M4 acute myeloid leukemia cell lines, belonging to the technical field of biochemistry and molecular biology. Background technique [0002] At present, more and more studies have found that cytokinins not only regulate the growth and development of plants, but also play an important role in the proliferation and growth of animal cells. Studies have found that some nucleoside forms of cytokinin ortho-TopolinRiboside (oTR), Kinetin riboside (KR), N 6 -Benzyladenosine (BAR), N6-Isopentenyladenosine (iPR) have strong toxicity and pro-apoptotic effects on cancer cells in vitro. Treatment of cancer cells with these cytokinins leads to cell cycle arrest and / or apoptosis to varying degrees, which mainly depends on the sensitivity of different cell lines to differe...

Claims

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Application Information

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IPC IPC(8): A61K31/7076A61P35/02C12N5/09C12Q1/02
Inventor 王黎刘胜先崔昌浩程娇潘慧
Owner DALIAN UNIV OF TECH
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