Applications of apigenin for preparation of medicines inhibiting liver cancer epithelial-mesenchymal transition

A technology for epithelial cells and liver cancer cells, which is applied in the field of medicine and can solve the problems of unexplained biological mechanism and no discovery of the inhibitory effect of apigenin on the epithelial cell-mesenchymal transition of liver cancer cells.

Inactive Publication Date: 2017-02-08
天承南运(天津)科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

That is to say, at the cognitive level of the prior art, it has not been found that apigenin has an inhibitory effect on the epith

Method used

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  • Applications of apigenin for preparation of medicines inhibiting liver cancer epithelial-mesenchymal transition
  • Applications of apigenin for preparation of medicines inhibiting liver cancer epithelial-mesenchymal transition
  • Applications of apigenin for preparation of medicines inhibiting liver cancer epithelial-mesenchymal transition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1 (apigenin detects the inhibitory effect on the proliferation of liver cancer cell lines Bel-7402 and PLC-PRF-5)

[0057] Experiment method: MTT colorimetric method

[0058] The specific experimental steps are as follows: different liver cancer cells were inoculated on 96-well culture plates, and the cell density was 5×10 3 pcs / well, then placed at 37°C, CO 2 (5%) overnight in the incubator. The next day, the culture medium was discarded, and cell culture medium containing 0, 25, 50, 75, 100, 125, 150, 175, and 200 μM apigenin were added respectively. Placed at 37°C, CO 2 (5%) After culturing in an incubator for 48 hours, add 20 μL of MTT (concentration: 5 mg / mL) to each well and continue culturing for 4 hours. Then the culture solution was sucked out, and 150 μ L DMSO was added to each hole as a solvent to dissolve formazan. After dissolving, the absorbance at 490 nm was measured with a microplate reader, and the cell survival rate was calculated accord...

Embodiment 2

[0061] Example 2 (in vitro cell level detection of apigenin on the migration ability of liver cancer cell lines Bel-7402 and PLC-PRF-5)

[0062] Experimental method: cell scratch method

[0063] The specific experimental steps are as follows: inoculate different liver cancer cells in 24-well plates and culture monolayer cells to 100%, use the tip of a sterile pipette tip to make a 100 μm scratch blank area, discard the liquid, wash twice with 1×PBS, and add the concentration 0, 10, 20μM apigenin cell culture solution, placed at 37°C, CO 2 (5%) Cultured in an incubator for 48 hours, placed on an optical microscope at a fixed position to take pictures at 0 hours, 12 hours, 24 hours and 48 hours respectively, and recorded the changes in the distance between cells at both ends of the scratch.

[0064] Relative starting position ratio (Ratio to 0h) = S xh / S 0h ; Use Excel software to draw the cell migration distance-time curve, and each index is represented by mean ± standard d...

Embodiment 3

[0067] Example 3 (in vitro cell level detection of apigenin on the impact of liver cancer cells Bel-7402 and PLC-PRF-5 invasion ability)

[0068] Experimental method: Transwell method

[0069] The specific experimental steps are as follows: on ice, mix the medium and matrigel 1:2, add 50 μL of matrigel dilution to the Transwell chamber, and after the matrigel solidifies, inoculate different liver cancer cells in the Matrigel-covered Transwell chamber, the cell density 2×10 4 cells / well, 200 μL of cell suspension in each well, and the concentrations of apigenin in the cell suspension were 0, 10, and 20 μM, respectively. The cells were then placed at 37°C, CO 2 (5%) Culture in the incubator for 24 hours, take out the cells after 24 hours, fix with cold methanol for 15 minutes, remove the matrigel and cells in the chamber with a cotton swab, then stain the cells on the bottom of the chamber with crystal violet for 20 minutes, wash the cells with PBS after staining 3 times to r...

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Abstract

The invention researches biological properties of apigenin based on experiment means, and innovatively found that the apigenin has a definite inhibiting effect for liver cancer epithelial-mesenchymal transition (EMT) and can invert EMT development processes. Molecular marking experiments show that the expression level of epithelial marker molecules in apigenin-interfered liver cancer cells increases, the expression amount of mesenchymal marker molecules decreases, and degrees of changes and apigenin are in a dose-dependent manner. Based on the abovementioned beneficial findings, the apigenin can be utilized to treat a plurality of diseases adopting the EMT as an effect. The inventor further found that a relieving effect of the apigenin on the EMT effect can be adopted as a treating path for liver cancer. Experiments prove that as the liver cancer cell EMT effect is effectively inverted, metastasis of liver cancer cells is significantly inhibited and liver cancer cell proliferation is limited, thus successfully proving the cancer treating function of the apigenin.

Description

technical field [0001] The invention relates to the technical field of medicine, and further relates to the new medical application of the substance, in particular to the application of apigenin in the preparation of liver cancer epithelial cell-mesenchymal transition inhibitory medicine. Background technique [0002] Epithelial-mesenchymal transition (EMT) refers to the biological process in which epithelial cells transform into mesenchymal phenotype cells through specific procedures. Plays an important role in embryonic development, chronic inflammation, tissue remodeling, cancer metastasis and a variety of fibrotic diseases, and its main features are decreased expression of cell adhesion molecules (such as E-cadherin), cytoskeletal transformation of cytokeratins Vimentin-based cytoskeleton and morphology have the characteristics of mesenchymal cells. Through EMT, epithelial cells lose their epithelial phenotypes such as cell polarity and connection with the basement memb...

Claims

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Application Information

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IPC IPC(8): A61K31/352A61P35/00A61P35/04A61P1/16
CPCA61K31/352
Inventor 杨诚孙涛周红刚
Owner 天承南运(天津)科技有限公司
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