Application of substance for inhibiting PHB gene in glioma treatment

A material and genetic technology, applied in drug combinations, antineoplastic drugs, medical preparations containing active ingredients, etc., can solve the problems of less than 5% survival rate of patients, easy recurrence, incomplete resection of lesions, etc.

Pending Publication Date: 2021-06-15
军事科学院军事医学研究院生物医学分析中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Surgery cannot completely remove the lesion, and radiotherapy and chemotherapy can inhibit tumor growth in a short period of time, but GBM is resistant to radiotherapy and chemotherapy and is prone to recurrence, so the 5-year survival rate of patients is still less than 5%

Method used

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  • Application of substance for inhibiting PHB gene in glioma treatment
  • Application of substance for inhibiting PHB gene in glioma treatment
  • Application of substance for inhibiting PHB gene in glioma treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1. Knockout of PHB gene can reduce the viability and self-renewal ability of glioma stem cells

[0068] 1. Knockout of PHB gene

[0069] 1. Knock out PHB gene using CRISPR / Cas9 gene knockout system

[0070] The PHB gene (full name: Prohibitin; Genebankaccession: NM_001281496, updated on January 3, 2021) (encoding PHB protein, NCBI ID: 5245, updated on January 3, 2021) was constructed using the CRISPR / Cas9 gene knockout system Two sgRNAs with different targets, namely target #1 (its targeting sequence is sequence 1 in the sequence listing), target #2 (its targeting sequence is sequence 2 in the sequence listing), negative control (Negative control, marked is Ctrl, and its target sequence is sequence 4). The PHB gene was then stably knocked out in cells 4121GSC and 3691GSC by lentiviral infection. Specific steps are as follows:

[0071] 1) Construction of Lenti-PHB-sgRNA plasmid

[0072] (1) Design and synthesis of PHB sgRNA primer sequences

[0073] Accordi...

Embodiment 2

[0141] Example 2. Knockout of PHB inhibits GBM tumor growth and prolongs survival of tumor-bearing mice

[0142] Immunodeficiency mice: Balb / c nude (Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.), 4 weeks old, 15-17 grams.

[0143] 1. In vitro pretreatment of cells

[0144] Cells: 4121GSC-ΔPHB-#1, 4121GSC-ΔPHB-#2, 4121GSC-Ctrl from Example 1 (control).

[0145]The immunodeficient mice were randomly divided into three groups, namely control group, knockout #1 group and knockout #2 group, with 6 mice in each group. The cells were diluted with NBM basal medium (Neurobasal Medium, Thermo Fisher, Cat. No. 12348-017) and injected into the right frontal lobe of the brain of immunodeficient mice, and each mouse was injected with 5 × 10 cells. 4 4121GSC-Ctrl, 4121GSC-ΔPHB-#1, 4121GSC-ΔPHB-#2 were injected into the control group, knockout #1 group and knockout #2 group, respectively.

[0146] On the 30th day of intracranial tumor formation (ie, the 30th day of cell inj...

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PUM

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Abstract

The invention discloses application of a substance for inhibiting a PHB gene in glioma treatment. The invention discovers that the knockout or knockdown of the PHB gene can significantly reduce the tumor cell sphere forming ability and cell viability of GSC, and can significantly reduce the clone forming ability of GSC cells, that is, the knockout or knockdown of the PHB gene can significantly inhibit the self-renewal ability of cells, but has no influence on the cell viability of common non-dry tumor cells. The experiment of the invention further proves that the knockout or knockdown of the PHB gene can inhibit the growth of GBM tumor and prolong the lifetime of tumor-bearing mice. The substance has wide application prospects.

Description

technical field [0001] The invention relates to the application of substances inhibiting PHB gene in the treatment of glioma in the field of biomedicine. Background technique [0002] Neuroepithelial tumors are collectively referred to as gliomas, which are the most common intracranial malignant tumors, accounting for about 40%-50% of brain tumors. It is divided into grades I to IV, from low to high. Brain gliomas can be divided into astrocytoma, medulloblastoma, glioblastoma multiforme (GBM), ependymoma, oligodendroglioma, etc. Glioblastoma (GBM) is a grade IV glioma, which is the most malignant tumor. Even after the most aggressive treatment, the average survival period of GBM patients is only 12-15 months. [0003] Brain glioma grows invasively and has no obvious boundary with normal brain tissue. GBM, as the most malignant glioma, infiltrates and metastasizes more rapidly, and is more likely to recur. The prognosis of patients is extremely poor. The clinical standard t...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61P35/00A61P25/00
CPCA61K45/00A61P35/00A61P25/00
Inventor 满江红周涛李爱玲黄浩浩韩秋影夏晴陈亮
Owner 军事科学院军事医学研究院生物医学分析中心
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