Use of rotenone as non-small cell lung cancer cell sensitizer

A non-small cell lung cancer, non-small cell technology, applied in the application field of rotenone as a non-small cell lung cancer cell sensitizer, can solve the problem of TRAIL resistance and other problems

Inactive Publication Date: 2010-12-08
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] In order to overcome the resistance of non-small cell lung cancer cells to TRAIL, the present invention provides a new...

Method used

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  • Use of rotenone as non-small cell lung cancer cell sensitizer
  • Use of rotenone as non-small cell lung cancer cell sensitizer
  • Use of rotenone as non-small cell lung cancer cell sensitizer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 The effect of rotenone on the death of non-small cell lung cancer cells (A549) induced by TRAIL. 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 have grown completely attached to the wall, add the following drugs: TRAIL 50ng / ml, TRAIL 100ng / ml, rotenone 10nM, TRAIL 50ng / ml+rotenone 10nM, TRAIL 100ng / ml+rotenone 10nM, rotenone 100nM, TRAIL 50ng / ml+rotenone 100nM, TRAIL100ng / ml+rotenone 100nM, rotenone 1000nM, TRAIL 50ng / ml+rotenone 1000nM, TRAIL100ng / ml+rotenone 1000nM, rotenone 10μM, TRAIL 50ng / ml+rotenone 10μM, TRAIL100ng / ml+rotenone 10μM, 3 wells in each group ( figure 1 , 2), the cells were collected after drug treatment for 12 hours, washed twice with pre-cooled PBS, added EGFP-labeled Annexin V (2μL), incubated on ice for 20min, quickly added PI (1μg / mL), an...

Embodiment 2

[0020] Example 2 Effects of TRAIL and rotenone on the activity of caspase 3 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 2moist 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, TRAIL 100ng / ml, TRAIL 100ng / ml+rotenone 1μM, rotenone 1μM, repeat 3 wells for each group ( image 3 ). 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.

[0021] Results: The activation of caspase 3 is an important indicator of cell apoptosis. Whether it is mediated by death receptors or mediated by mi...

Embodiment 3

[0022] Example 3 Toxicity test HEK293 and A549 cells were respectively 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 amount of cells 5×10 3 Each well was seeded in a 96-well cell culture plate. Add the drug TRAIL 100ng / ml+rotenone 1μM to treat for 12h, repeat 3 wells in each group ( Figure 4 ), after 12 hours of treatment, the cells were collected, and the apoptosis of A549 and HEK293 cells was detected according to Example 1.

[0023] Results: To investigate whether the combined synergistic effect of TRAIL and rotenone has toxic side effects on normal cells. This study compared the cytotoxicity of TRAIL100ng / ml+rotenone (1μM) on normal cell line (HEK293) and tumor cell line (A549). see results Figure 4 , in the case of obviously inducing apoptosis of A549 cells, TRAIL+r...

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Abstract

The invention relates to novel use of a mitochondrial respiratory chain inhibitor rotenone in reversing the tolerance of non-small cell lung cancer cells to targeted antineoplastic agents. The rotenone serves as a non-small cell lung cancer cell sensitizer and can reverse the tolerance of the non-small cell lung cancer cells to the targeted antineoplastic agents. The invention shows the combination of rotenone compounds and TRAIL has no obvious toxic or side effect on normal cells and is a novel medicinal composition used for treating the non-small cell lung cancer.

Description

technical field [0001] The invention relates to the new application of rotenone, a mitochondrial respiratory chain inhibitor, 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 ru...

Claims

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Application Information

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IPC IPC(8): A61K31/352A61P35/00
Inventor 殷武华子春施怡琳
Owner NANJING UNIV
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