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Application of As*O* combined with TSA in preparing medicament for inducing leukocythemia HL60 cell to convert from portion differentiation to terminal differentiation

A technology of TSA0.1 and TSA0.2, applied in the field of medicine and biology, can solve the problems of slow progress of induced differentiation therapy and difficulty in obtaining re-ease.

Inactive Publication Date: 2008-04-02
INST OF BASIC MEDICINE OF SAMS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although ATRA or As 2 o 3 The representative acute promyelocytic leukemia induced differentiation therapy has achieved great clinical success, while other types of leukemia induced differentiation therapy has made relatively slow progress
Most APL patients with successful induction of differentiation are difficult to achieve remission due to the formation of drug resistance

Method used

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  • Application of As*O* combined with TSA in preparing medicament for inducing leukocythemia HL60 cell to convert from portion differentiation to terminal differentiation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] (1). Inhibitory effect of TSA on the growth of HL-60 cells: set four different TSA concentration groups and TSA in 96-well plate Negative control group, three wells for each group, each well added 5 × 10 5 0.1ml / ml HL-60 cells. 37°C, 5% CO 2 After culturing for 0h, 24h, 48h, 72h, and 96h under the same conditions, add 10μl of 5mg / ml MTT solution to each well, continue to culture for 4h, centrifuge at 2000rpm for 10 minutes, discard the supernatant, add DMSO150μl / well, shake fully to dissolve the cells, add DMSO 150 μ l was used as a zero-adjusting hole, and a microplate reader was used to measure the absorbance A value at 570 nm and 630 nm wavelength, and calculate the cell proliferation inhibition rate (%)=(control group A value-experimental group A value) / control group A value×100%.

[0042] (2).As 2 o 3 Inhibitory effect on HL-60 cell growth: Take HL60 cells in the logarithmic growth phase, centrifuge and discard the supernatant, add fresh complete culture medium...

Embodiment 2

[0046] (1). Inhibitory effect of TSA on the growth of primary APL cells: set four different TSA concentration groups and TSA in 96-well plate Negative control group, three wells for each group, each well added 5 × 10 5 / ml primary APL cells (supplier: Shanghai Cell Bank, Chinese Academy of Sciences) 0.1ml. 37°C, 5% CO 2 After culturing for 0h, 24h, 48h, 72h, and 96h under the same conditions, add 10μl of 5mg / ml MTT solution to each well, continue to culture for 4h, centrifuge at 2000rpm for 10 minutes, discard the supernatant, add DMSO150μl / well, shake fully to dissolve the cells, add DMSO 150 μ l was used as a zero-adjusting hole, and a microplate reader was used to measure the absorbance A value at 570 nm and 630 nm wavelength, and calculate the cell proliferation inhibition rate (%)=(control group A value-experimental group A value) / control group A value×100%.

[0047] (2).As 2 o 3 Inhibitory effect on the growth of primary APL cells: take primary APL cells, centrifuge an...

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Abstract

The invention discloses the application of a As2O3 combined TSA in preparing drugs to induce the change of leukemia HL60 cells from partial differentiation to terminal differentiation. The concentration of the As2O3 combined TSA in preparing drugs to induce the change of leukemia HL60 cells from partial differentiation to terminal differentiation is: 0.25 to 0.5 Mu M of As2O3; 0.1 to 0.2 Mu M of TSA. In the aspect of drug combination to induce the change of leukemia HL60 cells from partial differentiation to terminal differentiation, the drug laid basis for clinical drug combination therapy to induce leukaemia differentiation.

Description

technical field [0001] The invention belongs to the field of medical biotechnology. especially involving an As 2 o 3 Application of combined TSA in preparation of medicine for inducing leukemia HL60 cells to transform from partial differentiation to terminal differentiation. Background technique [0002] The following titles involved in the present invention apply to the entire specification and claims: [0003] BSA: bovine serum albumin (Bovine Serum Albumin), Sigma company product; [0004] As 2 o 3 : arsenic trioxide (arsenic trioxide), product of Sigma Company; [0005] TSA: Trichostatin A (Trichostatin A), product of Sigma Company; [0006] MTT: Tetramethylazolium bromide [3-(4,5-dimthy-2-2thiazoly) 2,5-dipheny-2H-tetrazoliunbromide], product of Sigma; [0007] DMSO: Dimethyl Sulfoxide, product of Sigma; [0008] NBT: Nitroblue tetrazolium chloride, product of Sigma Company; [0009] PE-CD11b antibody: PE-labeled mouse anti-human CD11b monoclonal antibody was ...

Claims

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Application Information

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IPC IPC(8): A61K33/36A61K31/7048A61P35/02
Inventor 姜国胜温晓燕温培娥唐天华杨伟华任霞任海全李智慧
Owner INST OF BASIC MEDICINE OF SAMS
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