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Suppression of EMT mediated liver cancer metastasis by Arteannuin B and action mechanism research thereof

A liver cancer metastasis and mechanism of action technology, applied in the field of medicine, can solve problems such as low survival rate

Pending Publication Date: 2018-10-30
于荣敏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although eradicating tumors is the quickest way to treat liver cancer, it still has certain limitations. For example, most patients are accompanied by liver cirrhosis and / or low liver function. Not all patients with liver cancer can receive surgical treatment due to actual conditions such as the size of the tumor; In addition, even for patients who underwent surgery, the five-year survival rate after surgery was still very low

Method used

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  • Suppression of EMT mediated liver cancer metastasis by Arteannuin B and action mechanism research thereof
  • Suppression of EMT mediated liver cancer metastasis by Arteannuin B and action mechanism research thereof
  • Suppression of EMT mediated liver cancer metastasis by Arteannuin B and action mechanism research thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: TGF-β1 induces epithelial-mesenchymal transition in HepG2 cells

[0035] ①TGF-β1 induces the transformation of HepG2 cells from epithelial to mesenchymal morphology:

[0036] 1) HepG2 cells were placed in a cell culture incubator (37°C, 5% CO) with high-glucose DMEM medium (containing 100 U / mL penicillin, 0.1 mg / mL streptomycin and 10% fetal bovine serum). 2 ) static culture. Each well of the six-well plate was plated with HepG2 cells accounting for about 20% of the bottom area of ​​the well. After overnight adherence to the wall, fresh serum-free medium was replaced 6 hours before the experiment to obtain synchronized growth of the cells. Then, each well of the six-well plate was sequentially set as NC, 0.625ng / mL TGF-β1, 1.25ng / mL TGF-β1, 2.5ng / mL TGF-β1, 5ng / mL TGF-β1. Place the plate in a cell culture incubator (37°C, 5% CO 2 )to cultivate.

[0037] 2) The morphological changes of the cells were observed and photographed every 24 hours under an inver...

Embodiment 2

[0057] Example 2: Arteannuin B inhibits the proliferation of HepG2 cells

[0058] MTT experiment:

[0059] 1) Preparation of cell suspension: Discard the old medium, wash it once with PBS, add 1 mL of 0.25% trypsin to digest for 3 minutes, observe under the microscope that most of the cells become round, discard the trypsin, add 2 times the amount of trypsin to complete The digestion of the medium was terminated, and the cells in the bottle were gently blown down and transferred to a 15mL centrifuge tube, and centrifuged at 1000rpm for 5min.

[0060] 2) Discard the supernatant, add fresh medium, keep 1 / 3 of the cell suspension as seed, and keep the rest for later use.

[0061] 3) Cell counting and plating: After the cell suspension was blown and mixed, pipette 10 μL into a hemocytometer for counting under a microscope, and adjust the concentration of the cell suspension to 3×10 4 cells / mL. Add 100 μL of concentration-adjusted cell suspension to each well of a 96-well plate,...

Embodiment 3

[0068] Example 3: Inhibition of EMT-mediated migration of HepG2 cells by Arteannuin B

[0069] 1. Morphological observation of the transformation of HepG2 cells from epithelial to mesenchymal cells induced by Arteannuin B blocking TGF-β1:

[0070] 1) HepG2 cells were placed in a cell culture incubator (37°C, 5% CO) with high-glucose DMEM medium (containing 100 U / mL penicillin, 60 U / mL streptomycin and 10% fetal bovine serum). 2 ) static culture. Each well of the six-well plate was plated with HepG2 cells accounting for about 50% of the bottom area of ​​the well. After overnight attachment to the wall, fresh serum-free medium was replaced 6 h before the experiment.

[0071] 2) After the cells grow synchronously, add 2 mL of DMEM medium with a final concentration of 5 ng / mL TGF-β1 to the six-well cells of the six-well plate (group T), 2 mL of Arteannuin B containing TGF-β1 5 ng / mL at 10 μM 2 mL of DMEM medium, 2 mL of Arteannuin B 20 μM DMEM medium containing TGF-β1 5 ng / mL, ...

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Abstract

An Arteannuin B according to the invention is an artemisinin derivative which is separated from a traditional Chinese medicine sweet wormwood. The invention discloses application of the Arteannuin B in preparing a liver cancer cell metastasis preventing candidate medicament. An importance of the invention is discovery of an EMT mediated liver cancer metastasis suppressing function by the Arteannuin B. The Arteannuin B is characterized in that the Arteannuin B has a remarkable propagation suppression function on human liver cancer HepG2 cells, and can retard epithelial-mesenchymal transition (EMT) of the HepG2 cells through induction of the TGF-beta1 on the condition of a non-toxic dosage 40 [mu]m; wound healing test and Transwell migration test results prove a fact that the Arteannuin B remarkably suppress TGF-beta1-induced HepG2 cell metastasis function, and has high concentration dependence; the action mechanism research proves a fact that the Arteannuin B may retard EMT of the TGF-beta1-induced HepG2 cells through suppressing expression of p-Smad2, p-Smad3 and p-STAT3 and Twist1 of the Smad path. In-vitro tests prove a fact that the Arteannuin B has a good treatment function ona mice lung metastasis model. Therefore, the Arteannuin B can be researched and developed as a candidate medicament for suppressing human liver cancer cell metastasis.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to Arteannuin B inhibiting EMT-mediated liver cancer metastasis and its mechanism of action. Background technique [0002] According to the GLOBOCAN survey, there were 782,451 cases of liver cancer in 2012, accounting for 5.6% of the total number of cancers in the world during the same period, and 12.9% of the total number of cancers in China that year. The world standardized incidence rate (ASR-W) is 10.1 / 100,000. About 83% of liver cancers occur in underdeveloped regions, 50% of which occur in China. my country's cancer statistics from 2000 to 2011 show that the incidence of liver cancer ranks fifth or sixth among all cancers, and men are higher than women. In addition, liver cancer ranks second in male cancer mortality, second only to lung cancer, and ranks second or third in females. [0003] A variety of pathogenic factors can lead to liver cancer. Chronic hepatitis B or C v...

Claims

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Application Information

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IPC IPC(8): A61K31/365A61P35/04
CPCA61K31/365
Inventor 于荣敏成玘宋丽艳朱建华
Owner 于荣敏
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