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Signaling pathway inhibitor for up-regulating MLLT3 gene expression, drug and application of the signaling pathway inhibitor

A signaling pathway and gene expression technology, applied in the field of biomedicine, can solve problems that need to be studied

Pending Publication Date: 2021-04-30
SUSHENG BIOTECH (HAINAN) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the regulation of MLLT3 gene expression in hematopoietic stem cells remains to be studied

Method used

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  • Signaling pathway inhibitor for up-regulating MLLT3 gene expression, drug and application of the signaling pathway inhibitor
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  • Signaling pathway inhibitor for up-regulating MLLT3 gene expression, drug and application of the signaling pathway inhibitor

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Experimental program
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Effect test

Embodiment 1

[0029] CD34 isolated from umbilical cord blood + Hematopoietic stem and progenitor cells integrate the gene fragment cloned with the MLLT3 gene into the genome of the cell through a virus, allowing the MLLT3 gene to be expressed in the cell (OE-MLLT3), and contain SCF (stem cell growth factor), FLT-3 ligand, TPO thrombopoietic factor was cultured in Stemspan series medium for 7 days, and CD34 was detected by flow cytometry + CD45RA - Cell population, found CD34 in OE-MLLT3 experimental group + CD45RA - The proportion of the cell population was much higher than that of the control group, which was nearly 4 times that of the control group, indicating that MLLT3 had a positive effect on the maintenance and proliferation of hematopoietic stem cells in vitro.

Embodiment 2

[0031] CD34 isolated from umbilical cord blood + For hematopoietic stem and progenitor cells, DMSO (negative control group), SR1 (reported to be able to expand HSC), UM171 (reported to be able to amplify HSC) and JRS (containing JNK-IN-8, Rapamycin, SR1) small molecules were cultured for 7 days respectively, and the expression of MLLT3 in each group of cells was detected by qPCR technology.

[0032] Test results such as figure 2 As shown, the expression of MLLT3 in cells treated with UM171 and JRS was significantly increased compared with the control group, and JRS had a better effect than UM171 alone. However, SR1 alone had no significant effect on the expression of MLLT3 gene.

Embodiment 3

[0034] CD34 isolated from umbilical cord blood + Hematopoietic stem and progenitor cells were cultured for 7 days in the control group (Vehicle), MLLT3 overexpression group (OE-MLLT3) and experimental group JRS (medium containing JNK-IN-8, Rapamycin, and SR1 small molecules), and inoculated in the culture medium containing In the semi-solid medium of hemimethylcellulose, the number of GEMM colonies formed was observed under a microscope after 14 days.

[0035] Count results as image 3 As shown, the number of GEMM clones formed in the OE-MLLT3 and JRS experimental groups was significantly better than that in the control group.

[0036] Cells cultured for 7 days were inoculated in hemimethylcellulose semi-solid medium, and GEMM clones were observed under a microscope after 14 days, as shown in Figure 4 shown.

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Abstract

The invention discloses a signaling pathway inhibitor for up-regulating MLLT3 gene expression, a drug and application of the signaling pathway inhibitor. A JNK signaling pathway inhibitor, an optional mTOR signaling pathway inhibitor and an aromatic hydrocarbon receptor inhibitor act on the hematopoietic stem cells, MLLT3 gene expression can be effectively up-regulated, a reliable research means is provided for MLLT3 gene regulation and control, a foundation is laid for research and clinical application of hematopoietic stem cell function regulation and control through MLLT3. Important scientific research and clinical conversion values are realized.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a signal pathway inhibitor, a medicine for up-regulating the expression of MLLT3 gene in hematopoietic stem cells, and the application of the signal pathway inhibitor. Background technique [0002] Hematopoietic stem cells (HSCs for short) are a type of adult stem cells with high self-renewal ability and multi-lineage differentiation potential. According to literature reports, a single hematopoietic stem cell has the ability to rebuild the entire blood system, and can be used in the clinical treatment of various blood diseases such as leukemia, which has important application value. [0003] The MLLT3 transcription factor is an important member of the FOXO subfamily of the FOX family. It is a transcription factor that regulates various physiological functions such as cell oxidative stress response, cell proliferation, cell apoptosis, cell autophagy, metabolism and immune re...

Claims

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Application Information

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IPC IPC(8): C12N5/0789
CPCC12N5/0647C12N2501/999C12N2510/00
Inventor 曾胜陈鑫苹丰玫玫杨宗繁
Owner SUSHENG BIOTECH (HAINAN) CO LTD
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