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Application of cytarabine and proto-oncoprotein c-FOS inhibitor in preparation of product for treating leukemia

A cytarabine and leukemia technology, applied in the field of biomedicine, can solve the problems of high degree of tumor malignancy, recurrence, and drug resistance in children, and achieve the effect of overcoming drug resistance, increasing sensitivity, and reducing drug resistance.

Active Publication Date: 2020-12-01
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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AI Technical Summary

Problems solved by technology

Nevertheless, long-term and high-dose use of cytarabine is likely to cause drug-resistant reactions in children; in addition, children with high-risk subtypes of leukemia are prone to drug-resistant reactions and disease recurrence due to the high degree of tumor malignancy, which seriously affects the Efficacy and disease outcome of chemotherapy regimens for leukemia

Method used

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  • Application of cytarabine and proto-oncoprotein c-FOS inhibitor in preparation of product for treating leukemia
  • Application of cytarabine and proto-oncoprotein c-FOS inhibitor in preparation of product for treating leukemia
  • Application of cytarabine and proto-oncoprotein c-FOS inhibitor in preparation of product for treating leukemia

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1, the preparation of test cell and culture medium

[0045] 1. Collection, collection and processing of clinical samples

[0046] 1. Collection of clinical specimens

[0047] Bone marrow specimens from two newly diagnosed children with leukemia who were hospitalized in Beijing Children's Hospital during November 2018 were collected.

[0048] 2. Diagnosis and classification of newly diagnosed children with leukemia

[0049] Two children with leukemia were diagnosed as common B-ALL (common B lineage ALL, c-B-ALL) by clinical, morphological, cytochemical staining and immunological methods. Diagnosis and classification criteria were in accordance with national unified standards (Suggestions for Diagnosis and Treatment of Pediatric Acute Leukemia. Chinese Journal of Pediatrics. 1999,137(5):305-307). The multiplex nested reverse transcription-PCR method was used to detect 29 genes caused by chromosomal structural aberrations in the bone marrow specimens of the a...

Embodiment 2

[0093] Embodiment 2, the drug resistance detection of test cell and culture medium

[0094] 1. Detection of the apoptosis level of the stably transformed recombinant leukemia cells treated with chemotherapeutic drugs

[0095] Take the stably transformed recombinant leukemia cell Nalm-6 / BACH2 established in step 2 of Example 1 KD -2 with Nalm-6 / BACH2 Con As the test cells, they were cultured in 6-well plates containing 3 mL of RPMI-1640 medium containing 10% FBS, and the cell density was 5×10 5 To each well, add 20 nM of cytarabine (Ara-C), a chemotherapy drug commonly used in clinical practice for acute lymphoblastic leukemia (ALL), and normal saline (NS), harvest the cells after 48 hours, and analyze the cells by flow cytometry Apoptosis detection. The apoptosis detection kit is the 7-AAD / Annexin V apoptosis kit from BD Biosciences, USA. The result is as figure 2 shown.

[0096] The results showed that after treatment with Ara-C, the recombinant leukemia cells Nalm-6 / B...

Embodiment 3

[0103] Example 3, detection of leukemia cell survival rate and drug combination index treated with chemotherapy drugs

[0104] 1. Detection of the survival rate of leukemia cells treated with chemotherapy drugs

[0105] 1. Take the mononuclear cell samples Pt1 and Pt2 of the bone marrow of two cases of children prepared in the first step of Example 1, and the stable transfer recombinant leukemia cell line Nalm-6 / BACH2 established in the second step KD -2 and Nalm-6 / BACH2 Con As the test cells, they were cultured in 1 mL of 24-well plate containing common medium, and the cell density was 5×10 4 One / hole, each kind of test cell is divided into the following treatment groups according to the difference of adding drugs:

[0106] Treatment group 1: 20 nM cytarabine Ara-C.

[0107] Treatment group 2: 20 nM cytarabine Ara-C + 5 μM nordihydroguaiaretic acid (NDGA).

[0108] Treatment group 3: 20 nM cytarabine Ara-C + 5 μM curcumin.

[0109] Treatment group 4: 200 nM cytarabine Ar...

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Abstract

The invention discloses an application of cytarabine and a proto-oncoprotein c-FOS inhibitor in preparation of a product for treating leukemia. Experiments prove that a synergistic effect exists whencytarabine is combined with nordihydroguaiaretic acid or cytarabine is combined with curcumin to treat leukemia cells, and the drug-resistant reaction of leukemia cells in a cytarabine drug-resistantstrain cell and drug-resistant co-culture medium to cytarabine can be effectively reduced. The invention provides a new approach for overcoming drug resistance reaction for leukemia treatment, and theproto-oncoprotein c-FOS is expected to become a potential target for anti-leukemia treatment, and has very wide application prospect in the medical field.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of cytarabine and proto-oncoprotein c-FOS inhibitors in the preparation of products for treating leukemia. Background technique [0002] Activating protein-1 (activating protein-1, AP-1) is a heterodimer composed of c-JUN and c-FOS, which acts as a transcriptional activator in human cells. AP-1 responds to a variety of body stimuli by regulating gene expression, including cytokines, growth factors, stress, bacterial and viral expression, etc. Therefore, AP-1 is involved in the control of many cellular processes such as cell differentiation, proliferation and apoptosis. Numerous studies have found that AP-1 functions as a proto-oncoprotein in many human tumors. Nonetheless, c-JUN and c-FOS in AP-1 exert distinct functions by themselves. Among them, c-JUN is highly expressed in malignant tumors and participates in the occurrence and development of tumors by re...

Claims

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Application Information

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IPC IPC(8): A61K31/7068A61K45/06A61P35/02A61K31/05A61K31/12
CPCA61K31/7068A61K45/06A61P35/02A61K31/05A61K31/12A61K2300/00
Inventor 张寒张瑞东冯敏郑胡镛
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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