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Effect of artemisinin B in A549 cell migration and invasion inhibition

A technology of cell migration and artemisinin, which is applied in the field of medicine and can solve problems such as toxicity

Active Publication Date: 2017-01-04
于荣敏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research results of Huang et al. on dihydroartemisinin showed that dihydroartemisinin has selective cytotoxic effect on rat C6 cells by increasing reactive oxygen species and inhibiting the activation of hypoxia-induced related factors.

Method used

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  • Effect of artemisinin B in A549 cell migration and invasion inhibition
  • Effect of artemisinin B in A549 cell migration and invasion inhibition
  • Effect of artemisinin B in A549 cell migration and invasion inhibition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Effect of artemisinin B on colony formation of A549 cells

[0020] Take the monolayer cultured cells in the logarithmic growth phase, and prepare a cell suspension of 100 cells / mL. Cells were seeded in 6-well plates, 2 mL / well, 37°C, 5% CO 2 Incubate overnight in an incubator. Dosing culture: Discard the supernatant in the well, add artemisinin B liquid (0.625, 1.25, 2.5 and 5 μM) 2 mL / well, set 3 replicate wells for each concentration. The experiment set up a drug-dosing group and a negative control group, at 37°C, 5% CO 2 Continue culturing for 14 days under the same conditions, observe the cloning situation, and change the medium once during the period. After culturing for 14 days, the old culture medium in the culture plate was discarded, washed twice with PBS, and the cells were fixed with methanol for 30 min. Crystal violet staining was carried out for 30 min, and the excess staining solution was rinsed by soaking in tap water. Place the 6-well ...

Embodiment 2

[0022] Example 2: Effect of artemisinin B on the adhesion of A549 cells

[0023] Take monolayer cultured cells in the logarithmic growth phase, and prepare 10 5 cells / mL of cell suspension. Artemisinin B solutions were prepared with cell suspension, and the final concentrations were 40, 20, 10, and 5 μM, respectively. B cell suspensions containing different concentrations of artemisinin were inoculated in 6-well plates, 2 ml / well, with 3 replicate wells for each concentration. The experiment consisted of a dosing group and a negative control group. 37°C, 5% CO 2 After culturing for 2.5 h in an incubator, gently wash twice with PBS to remove unattached cells. Fix with methanol for 1 h, stain with crystal violet for 1 h, soak in tap water to rinse excess staining solution, take pictures and count.

[0024] The results showed that when the concentration of artemisinin B was 20 μM and 40 μM, the number of adhered cells was significantly less than that of the control group,...

Embodiment 3

[0025] Example 3 : Effect of artemisinin B on migration of A549 cells

[0026] ① Scratch healing experiment: draw a line on the bottom of the 6-well plate with a ruler and a marker pen, and divide the wells into 3 parts. Prepare 5×10 5 Cells / mL cell suspension, seeded in 6-well plate, 2 mL / well, confluent overnight. Scratch: Use a 10 μL pipette tip to make a scratch, the scratch passes through the three lines at the bottom, wash the cells twice with PBS, remove the scratched cells, and take pictures. Dosing culture: prepare 4 different concentrations of artemisinin B solutions (ie 40, 20, 10, 5 μM), 2 mL / well, and 3 replicate wells for each concentration, and set the dosing group and negative control group in the experiment. 37°C, 5% CO 2 Observed and photographed after 24 h in the incubator.

[0027] ② Transwell migration experiment: prepare 8×10 4 cells / 200 μl of cell suspension, the cell suspension was used to prepare different concentrations of artemisinin B soluti...

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Abstract

The present invention relates to artemisinin B, which has a molecular weight of 248 g / mol. The present invention discloses applications of the artemisinin B in preparation of drugs for tumor cell invasion and metastasis inhibition. According to the present invention, the key result comprises that the artemisinin B has the lung cancer A549 cell migration and invasion inhibition effect, wherein the effect specifically comprises that the artemisinin B provides the significant inhibition effect for the cloning proliferation of A549 cells; the artemisinin B inhibits the adhesive ability of A549 cells in the concentration-dependent manner; the results of the scratch test, the Transwell migration test and the Transwell invasion test show that the artemisinin B can effectively weaken the migration and invasion ability of A549 cells in the concentration-dependent manner; the microfilament fluorescence staining experiment results show that the artemisinin B inhibits the pseudopodia formation of A549 cells in the concentration-dependent manner; and the artemisinin B can significantly affect the invasion and metastasis-related proteins, and the experiment results show that the artemisinin B directly inhibits the STAT3 phosphorylation and the beta-catenin so as to down-regulate the expressions of MMP2 and MMP9, such that the artemisinin B has the application prospects in human lung cancer cell invasion and metastasis inhibition.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to the application of artemisinin B in inhibiting migration and invasion of A549 cells. Background technique [0002] Lung cancer seriously endangers human health and is one of the malignant tumors with the worst prognosis. Among them, non-small cell lung cancer (NSCLC) accounts for more than 80% of lung cancers. 70% of the deaths of lung cancer patients are caused by cancer metastasis. The mortality rate of lung cancer in my country is 27.93 / 100,000 (male 39.79 / 100,000, female 16.62 / 100,000), showing an increasing trend year by year. Lung cancer is one of the main malignant tumors affecting the health of Chinese residents. [0003] Lung cancer metastasis is the malignant sign and characteristic of lung cancer. It is the main cause of treatment failure and death of lung cancer patients. It is also the pathological basis of tumor recurrence, disease progression and eventual death o...

Claims

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Application Information

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IPC IPC(8): A61K31/365A61P35/04
Inventor 于荣敏刘婷宋丽艳
Owner 于荣敏
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