Application of induced differentiation reagent in M2 cells of human acute myeloid leukemia

An acute leukemia, cell technology, applied in animal cells, tumor/cancer cells, vertebrate cells, etc., can solve the problems of inappropriate ATRA induction therapy, instability, easy recurrence of patients, etc., and achieve significant in vitro proliferation inhibitory activity. Effect

Inactive Publication Date: 2018-11-23
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, ATRA-mediated differentiation therapy is not suitable for other types of AML, and ATRA induction therapy still has problems such as unstable curative effect and easy recurrence in some patients

Method used

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  • Application of induced differentiation reagent in M2 cells of human acute myeloid leukemia
  • Application of induced differentiation reagent in M2 cells of human acute myeloid leukemia
  • Application of induced differentiation reagent in M2 cells of human acute myeloid leukemia

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Effects of oTR, KR, BAR and iPR on the expression of CD11b in HL-60 cells: The resuscitated human M2 leukemia cells HL-60 were 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-h...

Embodiment 2

[0023] Morphological changes of HL-60 cells induced by oTR, KR, BAR and iPR: HL-60 cells in active growth state 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 HL-60 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 the application of an induced differentiation reagent in M2 type cells of human acute myeloid leukemia, and belongs to the technical field of biochemistry and molecular biology.The data disclosed by the invention are based on nucleoside forms of oTR, KR, BAR and iPR of cytokinin, and obvious in vitro proliferative inhibition activity on 9 major tumor lines and human 60 tumor cell lines, in particular to highest drug sensitivity to leukemia cell lines.

Description

technical field [0001] The present invention relates to cytokinin ortho-Topolin Riboside (oTR), Kinetin riboside (KR), N 6 - Benzyladenosine (BAR) and N6-Isopentenyladenosine (iPR), as a class of dedifferentiation-promoting reagents, have anti-tumor effects on human M2 type acute myeloid leukemia cell lines, and belong 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 o...

Claims

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Application Information

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IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2501/599
Inventor 王黎程娇崔昌浩刘胜先陈晨
Owner DALIAN UNIV OF TECH
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