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Application of uabain for enhancing cellular sensitivity of non-small cell lung cancer (NSCLC)

A non-small cell lung cancer, sensitive technology, applied in medical preparations containing active ingredients, organic active ingredients, peptide/protein ingredients, etc., can solve the problem of insufficient tumor cell selectivity, loss of the best time for surgery, normal tissue and cell damage

Inactive Publication Date: 2010-10-27
NANJING UNIV
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  • Description
  • Claims
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Problems solved by technology

And another 80% of patients with advanced lung cancer have lost the best opportunity for radical surgery
3: So far, the treatment of lung cancer, especially for patients with advanced lung cancer, lacks very effective therapeutic interventions
In fact, although some chemotherapeutic drugs can significantly reverse the sensitivity of non-small cell lung cancer cell line A549 to TRAIL, these chemotherapeutic drugs themselves are not selective enough for tumor cells, so they often cause damage to normal tissues and cells

Method used

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  • Application of uabain for enhancing cellular sensitivity of non-small cell lung cancer (NSCLC)
  • Application of uabain for enhancing cellular sensitivity of non-small cell lung cancer (NSCLC)
  • Application of uabain for enhancing cellular sensitivity of non-small cell lung cancer (NSCLC)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Annexin V / PI Flow Cytometry Detection of A549 Cell Apoptosis Caused by the Combination of TRAIL and Ouabain Non-small cell lung cancer cells A549 were cultured in DMEM+10% fetal bovine serum culture medium, the culture condition was 37°C, 5%CO 2 moist sterile environment. When the cells grow to 80%-90% confluent, they are washed twice with PBS and digested with 0.25% trypsin. According to the cell volume 6×10 4 Each well was seeded in a 24-well cell culture plate. After the cells adhere to the wall and grow completely, add the drugs respectively TRAIL 10ng / ml, TRAIL 10ng / ml+ouabain 100nM, TRAIL 20ng / ml, TRAIL 20ng / ml+ouabain 100nM, TRAIL 50ng / ml, TRAIL 50ng / ml+ouabain For 100nM, TRAIL 100ng / ml, TRAIL100ng / ml+ouabain 100nM, TRAIL 200ng / ml, TRAIL 200ng / ml+ouabain 100nM, TRAIL400ng / ml, TRAIL 400ng / ml+ouabain 100nM, 3 wells in each group ( figure 1 ), or TRAIL100ng / ml+ouabain 10nM, ouabain 10nM, TRAIL 100ng / ml+ouabain 20nM, ouabain 20nM, TRAIL100ng / ml+ouabain ...

Embodiment 2

[0023] Example 2 Fluorescence Microscopy Detection of A549 Cell Nuclear Fragmentation Caused by the Combination of TRAIL and Ouabain Non-small cell lung cancer cells A549 were cultured in DMEM+10% fetal bovine serum culture medium, and the culture conditions were 37°C, 5% CO 2 moist sterile environment. When the cells grow to 80%-90% confluent, they are washed twice with PBS and digested with 0.25% trypsin. According to the amount of cells 1×10 5 Each well was seeded in a 6-well cell culture plate. Pre-place the sterilized cover glass surface in the culture plate, and when the cells grow to 60% adherent, add drug treatment, respectively control group, TRAIL 100ng / ml, TRAIL 100ng / ml+ouabain 50nM, TRAIL 100ng / ml , TRAIL 100ng / ml+ ouabain 50nM, each group repeated 3 wells ( image 3 ). After 12 hours of treatment, the glass slides were taken out, washed twice with cold PBS, and a few drops of DAPI (1 μg / ml) were added, observed on a fluorescent microscope, and filmed.

[002...

Embodiment 3

[0025] Example 3 Effects of TRAIL and ouabain on the activity of caspase3 hydrolase Non-small cell lung cancer cells A549 were cultured in DMEM+10% fetal bovine serum culture medium, the culture conditions were 37°C, 5% CO 2 moist sterile environment. When the cells grow to 80%-90% confluent, they are washed twice with PBS and digested with 0.25% trypsin. According to the cell volume 6×10 4 Each well was seeded in a 24-well cell culture plate. Add drug treatment, respectively control group, TRAIL100ng / ml, TRAIL 100ng / ml+ouabain 50nM, TRAIL 100ng / ml, TRAIL 100ng / ml+ouabain 50nM, each group repeated 3 wells ( Figure 4 ). After 12 hours of treatment, the cells were digested with trypsin, 1-2 μl of FITC-labeled DEVD.fmk was added, incubated on ice for 30 minutes, washed twice with PBS, and the fluorescence intensity of FITC was analyzed by flow cytometry.

[0026] Results: When the cells are in the state of apoptosis, it will cause the activation of caspase3, so that many pro...

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Abstract

The invention relates to a new application of sodium-potassium ATP enzyme inhibitor uabain for reversing tolerance of cells of non-small cell lung cancer (NSCLC) to targeted anti-tumor drugs. Particularly, the uabain can be used as a sensitizer for treating the NSCLC and is combined with TRAIL to be used for treating the NSCLC.

Description

technical field [0001] The invention relates to a new application of sodium potassium ATPase inhibitor ouabain in reversing the resistance of non-small cell lung cancer cells to targeted antitumor drugs. Background technique [0002] Malignant tumors are major diseases that endanger human health. In recent years, Jiangsu Province has carried out a retrospective survey of the cause of death of the whole population, mainly malignant tumors. The results show that the first cause of death for both men and women is malignant tumors, and its mortality rate accounts for 27.41% of all causes of death. According to the data, in 2003, the top 4 malignant tumor deaths in urban areas of Jiangsu Province were lung cancer, gastric cancer, liver cancer, and esophageal cancer; while in rural areas, they were liver cancer, lung cancer, esophageal cancer, and gastric cancer. Lung cancer and gastric cancer mortality rates in urban areas are significantly higher than those in rural areas. In ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7048A61K38/17A61P35/00
Inventor 殷武华子春冯速
Owner NANJING UNIV
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