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Application of LukS-PV protein in preparation of medicine for inducing leukemia cell differentiation

A leukemia cell and protein technology, applied in the field of medicine, can solve the problems of retinoic acid syndrome, cytokine differentiation inducers, such as high price and tolerance, and achieve easy genetic modification and chemical modification, inhibit the growth of leukemia cells, reduce the Toxic effects

Inactive Publication Date: 2014-06-25
ANHUI PROVINCIAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Small molecule compounds have a single effect. For example, ATRA is mainly used for the treatment of acute promyelocytic leukemia, and the effect on other types of leukemia is poor, and some patients may experience retinoic acid syndrome, relapse after remission, and drug therapy during ATRA treatment. Tolerance and other issues
Cytokine differentiation inducers are expensive and difficult to use clinically

Method used

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  • Application of LukS-PV protein in preparation of medicine for inducing leukemia cell differentiation
  • Application of LukS-PV protein in preparation of medicine for inducing leukemia cell differentiation
  • Application of LukS-PV protein in preparation of medicine for inducing leukemia cell differentiation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] In this example, HL-60 cells in the logarithmic growth phase were taken, and the cell concentration was adjusted to 2×10 5 / ml, added to a six-well plate, 2×10 per well 5 cells. After HL-60 cells were treated with LukS-PV at concentrations of 0, 0.50, and 1.00 μM for 3, 6, 9, and 12 hours, the cell growth was detected by MTS method.

[0030] Such as figure 1 As shown, the results showed that the growth inhibition curve of the normal control group was horizontal, and no growth inhibition was observed. LukS-PV can significantly inhibit the growth of HL-60 cells in a time-dose-dependent manner, that is, the inhibitory effect is more obvious when the dose is increased and the time is prolonged.

[0031] The human promyelocytic leukemia cell line HL-60 cells in this example were purchased from the Shanghai Cell Institute of the Chinese Academy of Sciences.

Embodiment 2

[0033] In this example, the HL-60 cells in the logarithmic growth phase were used to adjust the cell concentration to 2×10 5 / ml, added to a six-well plate, 2×10 per well 5 cells. After the cells were treated with 0, 0.50 μM, and 1.00 μM LukS-PV for 48 hours, the cell suspension was centrifuged and smeared, and the morphological changes of the cells were observed under a light microscope.

[0034] Such as figure 2 As shown, the effect of LukS-PV on the morphology of HL-60 cells was observed by Wright-Giemsa staining, and the magnification was 400 times. The results showed that after LukS-PV was applied to the cells for 48 hours, the cell morphology changed significantly under the light microscope. It is characterized by reduced cell volume, disappearance of nucleolus, significantly reduced nucleus, increased cytoplasm, condensed and thickened chromatin, reduced ratio of nucleoplasm, and nuclei are biased to one side.

[0035] Other implementations are the same as in Exampl...

Embodiment 3

[0037] In this embodiment, the THP-1 cells in the logarithmic growth phase were used to adjust the cell concentration to 2×10 5 / ml, the THP-1 cells of the human monocytic leukemia cell line in this example were purchased from the Shanghai Cell Institute of the Chinese Academy of Sciences. Other implementations are the same as in Example 2.

[0038] Such as image 3 As shown, the effect of LukS-PV on the morphology of THP-1 cells was observed under a direct microscope, and the magnification was 400 times. After LukS-PV was applied to the cells for 48 hours, the cell morphology was directly observed under a light microscope. Significant changes occurred. It showed that the cells changed from round, suspended to spindle-shaped, adhered, and showed adherent growth.

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Abstract

The invention discloses an application of LukS-PV protein in preparation of a medicine for inducing leukemia cell differentiation. The leukemia cell refers to a human promyelocyte leukemia cell HL-60 or human monocyte leukemia cell THP-1. The LukS-PV protein can be massively expressed by using a genetic engineering technology, can be specifically combined with a target cell but not cause the dissolution of the target cell, is small in molecular weight, is easily modified genetically and chemically; and through gene modification, the induction activity is expected to be improved and the toxic and side effects are expected to be reduced; experiment shows that LukS-PV can induce the HL-60 cell to differentiate to marrow monocyte in vitro and induce the THP-1 cell to differentiate to macrophage; LukS-PV can inhibit leukemia cell growth in vivo and reduce infiltration of the leukemia cell to the liver, the spleen and the peripheral blood; therefore, the LukS-PV protein is expected to be developed into a novel leukemia treatment medicine, thereby opening up a novel application field for bacterial toxin.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to the application of a LukS-PV protein in the preparation of drugs for inducing differentiation of leukemia cells. Background technique [0002] Leukemia is a systemic disease and thus cannot be treated with surgery or radiation like other solid tumors. Chemotherapy is the main method of treating leukemia. However, due to the non-specific cytotoxicity and inevitable drug resistance of chemotherapy drugs, most patients will eventually die from treatment-related coagulation disorders, bacterial infections, or disease recurrence caused by drug resistance. Exploring new targeted chemotherapeutic drugs for leukemia is a hot spot in clinical medical research. [0003] The molecular biological mechanism of leukemia pathogenesis is closely related to abnormal cell differentiation. Inducing differentiation is a new strategy for the treatment of leukemia. The difference between this meth...

Claims

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Application Information

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IPC IPC(8): A61K38/16A61P35/02
Inventor 马筱玲常文娇单武林章成芳
Owner ANHUI PROVINCIAL HOSPITAL
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