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A targeted inhibitor of dbf4p1 gene and its use

An inhibitor and targeting technology, applied in genetic engineering, medical preparations containing active ingredients, recombinant DNA technology, etc., can solve the problem of unclear regulation of pseudogene DBF4P1, the expression level and mechanism of action of pseudogene DBF4P1, etc. problems, achieving low off-target efficiency, solving drug resistance problems, and high silencing efficiency

Active Publication Date: 2022-02-22
SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are no relevant research reports on the expression level and mechanism of the pseudogene DBF4P1 in malignant tumors
Meanwhile, the regulatory role of the pseudogene DBF4P1 in glioblastoma remains unclear

Method used

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  • A targeted inhibitor of dbf4p1 gene and its use
  • A targeted inhibitor of dbf4p1 gene and its use
  • A targeted inhibitor of dbf4p1 gene and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0032] Example 1 Design of shRNA and preparation of corresponding interference vector and application of DBF4P1 gene targeting inhibitor.

[0033] 1. Cell culture.

[0034] Normal human brain astrocytes (NHA) and human glioblastoma cell lines A172, U87, U251 and U373 used in the study were purchased from the Life Science Cell Resource Center of Shanghai Research Institute. Culture NHA, A172, U87, U251, and U373 cells in 100mm cell culture dishes using DMEM high-glucose medium supplemented with 10% fetal bovine serum, add 6ml of the above culture medium to each culture dish, and wash with PBS every 2 days , and replace the culture medium at the same time, generally the cells can grow to a monolayer in about 2-3 days.

[0035] 2. Real-time fluorescent quantitative PCR technique was used to detect the expression level of DBF4P1.

[0036] 1. Trizol method to extract total RNA in cells.

[0037]Discard the cell culture medium, wash with pre-cooled PBS for 3 times, add 1ml Trizol...

Embodiment 2

[0051] Example 2 Application of DBF4P1 gene-targeted inhibitor significantly inhibits biological behaviors such as proliferation, migration and invasion of glioblastoma.

[0052] 1. CCK-8 cell viability assay was used to detect the effect of DBF4P1 gene targeting inhibitor on the proliferation ability of glioblastoma cells.

[0053] The CCK8 kit was used to detect the cell proliferation ability, and the cells were digested with trypsin to make a single cell suspension. For cell counting, cells in different groups were inoculated and cultured in 96-well plates at a density of 2000 cells per well (three replicate wells for each group). The 96-well plate after seeding the cells was placed in a constant temperature cell incubator for 24 hours (37°C, 5% CO 2 ), after adding 10ul of CCK-8 reagent, use a microplate reader to detect the light absorption value of each inoculated well at 450nm OD value at different time points of 30min, 60min, 90min, and 120min. After using the DBF4P1...

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Abstract

The invention belongs to the technical field of biomedicine, and in particular relates to a targeted inhibitor of DBF4P1 gene and its application. A targeted inhibitor of the DBF4P1 gene has the effect of inhibiting the expression of the DBF4P1 gene, and the target sequence of the inhibitor is: 5'‑GATATAACGGTGATAGAGCAG‑3' (SEQ ID No.1). Application of the DBF4P1 gene targeting inhibitor in the preparation of drugs for targeted treatment of human brain glioblastoma. The present invention develops a targeted inhibitor of the DBF4P1 gene through RNA interference technology, and the targeted inhibitor can specifically bind the DBF4P1 gene to play a role in inhibiting the expression of the DBF4P1 gene, thereby inhibiting the proliferation and migration of glioblastoma cells and invasion, thereby playing a targeted therapeutic role in inhibiting the growth of glioblastoma. The application of DBF4P1 gene targeting inhibitor can play an important role in the field of glioblastoma gene therapy, and provide new molecular targeted therapy drugs for clinical glioblastoma gene therapy.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a targeted inhibitor of DBF4P1 gene and its application. Background technique [0002] Glioblastoma is the most common and most malignant primary tumor of the central nervous system, with rapid proliferation and strong invasiveness. Surgical treatment combined with radiotherapy and chemotherapy, as well as anti-angiogenic therapy and other comprehensive treatment methods have become an important means for the treatment of glioblastoma at this stage, but the treatment effect is still not satisfactory, the recurrence rate of patients is high, the mortality rate is high, and the median survival rate is The period is only about 15 months. Recent studies have found that pseudogenes with similar functions to non-coding RNAs are abnormally expressed and play an important regulatory role in various malignant tumors. Among them, the research on the mechanism of pseudogen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113A61K31/713A61P35/00
CPCC12N15/1135A61P35/00C12N2310/14
Inventor 刘啸白刘云会薛一雪杨春清喻博李振商秀丽王振华王迪阮雪蕾李志清郑健奚卓腾浩赵德丰
Owner SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
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