Application of cytokinin in causing of human leukemia cell apoptosis through nucleoside transporter

A nucleoside transporter, leukemia cell technology, applied in medical preparations containing active ingredients, organic active ingredients, drug combinations, etc.

Inactive Publication Date: 2018-02-02
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The mechanism of cytokinin oTR transmembrane transport into cells has not been reported yet

Method used

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  • Application of cytokinin in causing of human leukemia cell apoptosis through nucleoside transporter
  • Application of cytokinin in causing of human leukemia cell apoptosis through nucleoside transporter
  • Application of cytokinin in causing of human leukemia cell apoptosis through nucleoside transporter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Ara-C and oTR inhibit the in vitro proliferation of human acute myeloid leukemia cells THP-1: culture the resuscitated human leukemia cells THP-1 in 10% FBS inactivated fetal bovine serum, 1% penicillin / 1% streptomycin (Double antibody) sterile DMEM culture medium, placed in a cell culture flask, at 37 ℃, 5% CO 2 And cultured in an incubator with a relative humidity of 90%, the cells grow to about 70%-80% for one passage. When THP-1 cells grow well, centrifuge at 1000 rpm / min for 5 minutes, collect cells in logarithmic growth phase, and dilute with DMEM medium to a cell density of 5×10 4 / ml, 100μl per well was inoculated into a 96-well plate. According to the experimental requirements, the treatment group was added with 0.1% DMSO, 0.3μM, 1.5μM, 15μM, 30μM concentration of Ara-C or oTR, 5 replicate wells each, and placed Incubate in an incubator for 3 to 5 days. Use trypan blue staining to detect the ratio of live cells to dead cells, and calculate the percentage of cell...

Embodiment 2

[0026] Transporter antagonist reverses the in vitro proliferation inhibition of human acute myeloid leukemia cells THP-1 by oTR: Cultivate THP-1 cells in good growth condition, centrifuge at 1000 rpm / min for 5 min, collect cells in logarithmic growth phase, and dilute with DMEM medium To a cell density of 5×10 4 / ml, 100μl per well was inoculated into 96-well plate, and A 1 , A 2A , A 2B , A 3 After incubating with adenosine receptor antagonist (DPCPX, SCH58261, MRS1754, MRS1191) and 5μM Dipyridamole (nucleoside transporter antagonist) for 2h, add 1.5μM oTR and place at 37℃, CO 2 Cultivate in an incubator for 4 days. Use CCK-8 kit for detection, add 10μl CCK-8 solution to each well of cell suspension, place at 37℃, CO 2 Incubate in an incubator for 2 hours, measure the absorbance of each group of cells at 450 nm, and record the data. Such as image 3 As shown, the combined effect of the four adenosine receptor antagonists (DPCPX, SCH58261, MRS1754, MRS1191) and 1.5μMoTR in Figu...

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Abstract

The invention provides an application of cytokinin in causing of human leukemia cell apoptosis through a nucleoside transporter, and belongs to the technical fields of biochemistry and molecular biology. Data in the invention is established on the basis of oTR having very significant in-vitro propagation inhibition activity to sixty tumor cell lines in nine human tumor systems and having strongestdrug susceptibility to leukemia cell strains. The data adopts the basic chemical structure of cytokinin oTR, the oTR is a nucleoside analog, the surface of a cell membrane has various molecule channels, and the researches of the oTR passing through the molecular channels and entering the cells provide a basis for the study of the antitumor effect.

Description

Technical field [0001] The present invention relates to the anti-tumor effect of cytokinin ortho-Topolin Riboside (oTR) on human leukemia cell lines. Transporter antagonists can reverse the anti-tumor effect of cytokinin oTR, belonging to the technical fields of biochemistry and molecular biology. Background technique [0002] Leukemia is a type of malignant clonal disease of bone marrow, also known as "blood cancer", including chronic myeloid leukemia, chronic lymphocytic leukemia, acute lymphoblastic leukemia and acute myeloid leukemia Leukemia (Acutemyeloid leukemia) is currently incurable. The prognosis of a patient largely depends on the type and age of the leukemia. Among them, acute myeloid leukemia is the type of acute leukemia with the highest incidence in adults, characterized by rapid onset and rapid development of the disease. Patients' complete remission rate and long-term disease-free survival rate are both low. [0003] At present, clinically, cytarabine (Ara-C) i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7076A61P35/02
Inventor 崔昌浩张瀚文王黎程娇林凡琳邓营营潘慧刘胜先
Owner DALIAN UNIV OF TECH
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