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Application of reagent for inhibiting or down-regulating expression of AKAP12 gene in preparation of tumor radiotherapy sensitizing drugs

A tumor radiotherapy and gene expression technology, applied in the direction of anti-tumor drugs, drug combinations, pharmaceutical formulations, etc., can solve the problems of no one reporting

Active Publication Date: 2018-08-31
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, the application of reagents that inhibit or down-regulate the expression of AKAP12 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 AKAP12 gene in preparation of tumor radiotherapy sensitizing drugs
  • Application of reagent for inhibiting or down-regulating expression of AKAP12 gene in preparation of tumor radiotherapy sensitizing drugs
  • Application of reagent for inhibiting or down-regulating expression of AKAP12 gene in preparation of tumor radiotherapy sensitizing drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] (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, passaged every 2-3 days, and cells in logarithmic growth phase were used for experiments.

[0032] (2) Cell transfection: One day before transfection, cells were digested with trypsin, counted, seeded in a 6-well plate at a suitable density, and placed in CO. 2 Incubate overnight in an incubator, so that the cell confluence on the day of transfection is 70% to 90%. Avoid antibiotics during plating and transfection.

[0033] Preparation of transfection solution: According to the dosage ratio of plasmid: Lipofectamine3000, 1 μg / well: 1.5 μl / well. Add 500ng / well DNA and 5μl / well P3000 to 250μl Opti-MEM medium and mix, and let stand for 5 minutes at room temperature for later use; 3.75μl / well Lipofectamine3000 is added to 250μl / well Opti-MEM medium, mix well, and let stand at room temperature for 5 minutes for later ...

Embodiment 2

[0036] (1) cell culture and cell transfection are the same as in Example 1;

[0037] (2) Cell clone formation method: take logarithmic growth phase A549 cells and A549 cells transfected with AKAP12siRNA and NC sequence for 72h, and inoculate different numbers of cells into six-well plates according to different irradiation dose requirements (0, 2, 4, The number of cells seeded with 8Gy dose was 200, 400, 800 and 1600, respectively). The three groups of cells were simultaneously irradiated with different doses (0-8Gy) of γ-rays, and the culture was terminated when the cell clones that were visible to the naked eye appeared in the culture dish. The culture medium was discarded, washed twice with PBS, fixed with anhydrous methanol for 30 minutes, stained with Gimsa stain for 30 minutes, washed with running water, air-dried, and counted to calculate the cell viability. The result is as figure 2 As shown, with the increase of irradiation dose, the cell death rate of A549 cells t...

Embodiment 3

[0039] (1) cell culture and cell transfection are the same as in Example 1;

[0040] (2) Detection of apoptosis by flow cytometry: A549 cells in logarithmic growth phase and A549 cells transfected with AKAP12siRNA and NC sequence for 72h (1×10 5 / mL) were seeded into six-well plates, and three groups of cells were given 8Gy 60 Coγ-ray irradiation, and then continued to culture for 24 hours, the cells were digested with trypsin without EDTA, centrifuged at 1500 rpm for 5 minutes, washed three times with PBS, and double-stained with Annexin V-FITC and PI to detect cell apoptosis by flow cytometry . The result is as image 3 As shown, the apoptosis rate of A549 cells transfected with AKAP12siRNA was significantly higher than that of normal A549 cell group and A549 group transfected with NC sequence.

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Abstract

The invention relates to the field of gene medicine, and in particular, relates to an application of a reagent for inhibiting or down-regulating the expression of an AKAP12 gene in preparation of tumor radiotherapy sensitizing drugs. The invention provides the AKAP12 gene and the new application of the reagent for inhibiting or down-regulating the expression of the AKAP12 gene, and the radiosensitivity of tumor cells can be effectively improved.

Description

technical field [0001] The invention relates to the field of gene medicine, in particular to the application of a reagent for inhibiting or down-regulating the expression of AKAP12 gene in the preparation of a sensitizing drug for tumor radiotherapy. Background technique [0002] Lung cancer is one of the most common tumors in my country. According to the results of the survey on the cause of death of Chinese residents, the mortality rate of lung cancer increased by nearly 1.5 times in the 20 years from the mid-1970s to the early 1990s, making it the fastest growing malignant tumor. Lung cancer generally refers to cancer of the lung parenchyma, usually excluding other mesodermal tumors of pleural origin, or other malignancies such as carcinoids, malignant lymphomas, or tumors that metastasize from other sources. Lung cancer accounts for 90-95% of lung parenchymal malignancies. In the past 50 years, the incidence and mortality of lung cancer in many countries in the world ha...

Claims

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Application Information

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IPC IPC(8): A61K31/7088A61K31/713A61P35/00
CPCA61K31/7088A61K31/713A61P35/00
Inventor 陈媛媛蔡建明高福杨彦勇雷霄郭佳铭李百龙崔建国刘哲许洋刘蕾曲红金
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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