Novel method for inhibiting colon cancer cell activity by combining inhibitor with small interfering RNA

A colon cancer cell and inhibitor technology, applied in the field of biomedicine, can solve the problem of aggravating DNA oxidative damage in cancer cells, achieve strong anti-colon cancer cell activity, weak inhibitory effect, and increase the level of active oxygen

Pending Publication Date: 2021-01-01
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Interfering the expression of SLC12A8 gene with small interfering RN

Method used

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  • Novel method for inhibiting colon cancer cell activity by combining inhibitor with small interfering RNA
  • Novel method for inhibiting colon cancer cell activity by combining inhibitor with small interfering RNA
  • Novel method for inhibiting colon cancer cell activity by combining inhibitor with small interfering RNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The required experimental materials and reagents of embodiment 1, embodiment 2, embodiment 3, embodiment 4 are as follows:

[0023] Experimental cell lines: human colon cancer cells HT29 and HCT116, human colorectal adenocarcinoma cells CACO2, human normal colon epithelial cells NCM460.

[0024] Chemicals and reagents: Oxaliplatin, Annexin V-FITC / PI Apoptosis Detection Kit, Cell Cycle and Apoptosis Detection Kit, genOFF st-h-SLC12A8_001.

[0025] Biologics: Olaparib, FK866

[0026] Example 1

[0027] MTT assay was used to detect the effects of different drug combinations on cell viability.

[0028] Adjust cell concentration to 2x10 with complete medium 4 / ml, seeded in 96-well plate, 200ul per well, cultivated overnight, and treated the cells with different drug combinations the next day. The drug combinations are: FK866, olaparib, FK866+olaparib, oxaliplatin, siSLC12A8, siSLC12A8+FK866, siSLC12A8+olaparib, siSLC12A8+FK866+olaparib, siSLC12A8+oxaliplatin. At the sa...

Embodiment 2

[0032] Cell scratch experiment (WoundHealing), to detect the effect of different drug combinations on cell migration.

[0033] First use a marker pen on the back of the 6-well plate, draw a horizontal line evenly with a ruler, about every 1cm, across the hole. Pass at least 5 wires through each hole. Adjust cell concentration to 5x10 with complete medium 5 / ml, inoculated in 6-well plates, 2ml per well, and cultivated overnight. The next day, use the tip of the gun to compare with a ruler, and try to hang down to the horizontal scratches on the back. The tip of the gun should be vertical and not tilted. Wash cells 3 times with PBS to remove scratched cells. Add different drug combinations, the drug combination is oxaliplatin and siSLC12A8+FK866+olaparib. At the same time, an equal volume of complete medium was used as the control group. Put into 37 degree 5% CO2 incubator, culture. Take samples at 0, 12, and 24 hours and take pictures.

[0034] Effects of different drug...

Embodiment 3

[0036] The effects of different drug combinations on cell apoptosis were detected by flow cytometry.

[0037] Annexin V-FITC / PI Cell Apoptosis Detection Kit respectively detects the effects of oxaliplatin and siSLC12A8+FK866+olaparib on cell apoptosis.

[0038] Effects of different drug combinations on apoptosis such as image 3 As shown, both drugs can promote cell apoptosis, and siSLC12A8+FK866+olaparib has a stronger ability to promote cell apoptosis.

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Abstract

The invention relates to a novel method for inhibiting colon cancer cell activity by combining an NAMPT inhibitor and a PARP inhibitor with small interfering RNA. The novel method has the advantages that an anti-tumor molecular targeting drug and an RNA interference technology are creatively combined, and tumor cells are specifically dead by utilizing the molecular biological difference between the tumor cells and normal cells. According to the novel method, drug combination design is carried out according to the characteristics of high expression of NAMPT in colon cancer cells and high sensitivity to oxidative damage, and compared with traditional chemotherapeutic drugs, the obtained combined drug combination method has higher anti-colon cancer cell activity and weaker inhibition effect on normal colon epithelial cell activity, and has higher biological safety in the aspect of cell level.

Description

technical field [0001] The invention relates to the field of biomedicine, and is a new method for inhibiting the activity of colon cancer cells by combining NAMPT inhibitors and PARP inhibitors with small interfering RNAs. Background technique [0002] Colon cancer is the third most common cancer in humans and one of the cancers with the highest mortality rate. There are approximately 1.2 million new cases of colon cancer worldwide each year. With the development of my country's economy, great changes have taken place in people's diet structure and living habits. The incidence of colon cancer in my country is increasing year by year, ranking third among all malignant tumors, with a mortality rate of 0.01%. [0003] Oxaliplatin (Oxaliplatin) is one of the first-choice drugs for the treatment of colon cancer. It acts on DNA by producing alkylated conjugates to form intra-strand and inter-strand crosslinks, thereby inhibiting DNA synthesis and replication and exerting anti-tumo...

Claims

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Application Information

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IPC IPC(8): A61K31/713A61P35/00A61K31/4545A61K31/502
CPCA61K31/4545A61K31/502A61K31/713A61P35/00A61K2300/00
Inventor 马文建周飒刘心怡蔡玉巧宋玉鹏聂志岩
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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