Application of reagent for inhibiting or down-regulating expression of GTSE1 gene in preparation of tumor radiotherapy sensitization drug

A technology of gene expression and tumor radiotherapy, applied in the field of biomedicine, can solve the problems of no one reporting and achieve the effect of improving sensitivity, increasing radiation sensitivity, and promoting DNA damage

Pending Publication Date: 2020-07-28
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether tumor cell resistance to chemotherapy is also related to the abnormally high expression of GTSE1 is unknown. Therefore, the application of inhibiting or down-regulating GTSE1 gene in the preparation of tumor radiosensitizing drugs has not been reported at home and abroad.

Method used

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  • Application of reagent for inhibiting or down-regulating expression of GTSE1 gene in preparation of tumor radiotherapy sensitization drug
  • Application of reagent for inhibiting or down-regulating expression of GTSE1 gene in preparation of tumor radiotherapy sensitization drug
  • Application of reagent for inhibiting or down-regulating expression of GTSE1 gene in preparation of tumor radiotherapy sensitization drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1, GTSE1 siRNA transfection

[0032] Transfect GTSE1 siRNA into lung cancer cells A549, H460, and H1299, taking transfection of A549 cells as an example, the cell transfection process is as follows:

[0033] (1) Cell culture: A549 cells were cultured in DMEM medium containing 10% fetal bovine serum; the cells were placed at 37°C, 5% CO 2 Cultured in an incubator, subcultured every 2-3 days, and the cells in the logarithmic growth phase were used for experiments.

[0034] (2) Cell transfection: The day before transfection, the cells were digested with trypsin, counted, inoculated in a 6-well plate at an appropriate density, and placed in CO 2 Cultivate overnight in an incubator, so that the confluence of the cells on the day of transfection is 70%-90%. Avoid antibiotics during plating and transfection.

[0035] Preparation of transfection solution: plasmid: Lipofectamine3000 dosage ratio was 1 μg / well: 1.5 μl / well. Add 500ngDNA and 5μL LP3000 to each well,...

Embodiment 2

[0038] Example 2, GTSE1 siRNA enhances the radiation sensitivity of lung cancer cells

[0039] (1) Cell culture and cell transfection method are the same as embodiment one;

[0040] (2) Cell clone formation method:

[0041] Take lung cancer A549, H460, H1299 cells in the logarithmic growth phase and A549, H460, H1299 cells transfected with GTSE1siRNA for 72 hours, and inoculate different numbers of cells into six-well plates according to different irradiation dose requirements (0,2,4, The number of cells inoculated with 8Gy dose was 200, 400, 800 and 1600, respectively).

[0042] The three groups of cells were irradiated with different doses (0-8Gy) of γ-rays at the same time, and continued to culture after irradiation until obvious cell clones visible to the naked eye appeared in the culture dish, and the culture was terminated. The medium was discarded, washed twice with PBS, fixed with anhydrous methanol for 30 minutes, stained with Gimsa stain for 30 minutes, rinsed with...

Embodiment 3

[0044] Example 3, GTSE1 siRNA promotes apoptosis of lung cancer cells

[0045] (1) Cell culture and cell transfection are the same as in Example 1;

[0046] (2) Cell apoptosis detection by flow cytometry: Take A549, H460, H1299 cells in the logarithmic growth phase and A549, H460, H1299 cells transfected with GTSE1siRNA for 72 hours, respectively, with 1 × 10 5 / mL concentration was inoculated into six-well plates, and each group of cells was given 8Gy 60 Coγ-ray irradiation, and then continue to culture for 24 hours, use EDTA-free trypsin to digest cells, centrifuge at 1500 rpm for 5 minutes, wash with PBS three times, use Annexin V-FITC and PI double staining to detect cell apoptosis by flow cytometry .

[0047] The result is as image 3 As shown, the apoptosis rate of A549, H460, H1299 cells transfected with GTSE1siRNA for 72 hours was significantly higher than that of normal A549, H460, H1299 cell groups.

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Abstract

The invention relates to the technical field of biomedicine engineering and provides application of a reagent for inhibiting or down-regulating expression of a GTSE1 gene in preparation of a tumor radiotherapy sensitization drug, particularly application in preparation of a lung cancer radiotherapy sensitization drug. Experiments prove that GTSE1 knockdown promotes DNA damage of exposure to lung cancer cells, increases the radiosensitivity of three lung censer cells significantly and increases the apoptosis rate of the three lung cancer cells after exposure significantly with the increase of an exposure dose, and the cell survival rate of the three lung cancer cells is obviously lower than that of a normal lung cancer cell group. Therefore, the application provides a new exploration for improving the sensitivity of radiotherapy, contributes to radiotherapy sensitivity of malignant tumor and has certain clinical application prospects.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of a reagent for inhibiting or down-regulating GTSE1 gene expression in the preparation of tumor radiosensitizing drugs, a carrier and a drug containing the same. Background technique [0002] Lung cancer is the malignant tumor with the highest morbidity and mortality rate in the world, accounting for 30-40% in China and other developing countries. In the past 40 years, the incidence of lung cancer in my country has been increasing year by year. In 2015, the number of lung cancer cases in China reached about 781,000, ranking first in the incidence of malignant tumors in my country. Lung cancer has an insidious onset and high degree of malignancy. The five-year survival rate of patients is extremely low. It has become one of the malignant tumors that seriously threaten human health. According to the data of the World Health Organization, lung cancer is current...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K31/7105A61P35/00
CPCA61K48/005A61K48/0008A61K31/7105A61P35/00
Inventor 曲宝林雷霄马娜杜乐辉俞伟张沛解传滨韩亚楠梁延杰
Owner GENERAL HOSPITAL OF PLA
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