Vitro model capable of Dynamic modeling human cancer of liver displace

A dynamic simulation, liver cancer metastasis technology, applied in animal cells, vertebrate cells, tumor/cancer cells, etc., can solve the problem that liver cancer metastasis model is not considered.

Inactive Publication Date: 2008-05-21
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this factor has not been taken into account in the current liver cancer metastasis model.
After searching, no papers or patents have been reported on the construction of an in vitro model that dynamically simulates human liver cancer metastasis and the effective use of an in vitro model that dynamically simulates human liver cancer metastasis to dynamically simulate and systematically study the biological activity of metastatic liver cancer cells

Method used

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  • Vitro model capable of Dynamic modeling human cancer of liver displace
  • Vitro model capable of Dynamic modeling human cancer of liver displace
  • Vitro model capable of Dynamic modeling human cancer of liver displace

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Construction of an in vitro model for dynamically simulating liver cancer metastasis process

[0042] 1. Model establishment of anti-cell attachment

[0043] Adherent growth is a prerequisite for the survival of solid tissue cells. Poly-HEMA (purchased from Sigma Company), as an anionic polymer, can be spread on the bottom of the container to prevent the cells from adhering to the wall, so as to construct a cell model in which cells grow in suspension after shedding.

[0044] Dissolve poly-HEMA with 95% ethanol to prepare a solution with a final poly-HEMA concentration of 120mg / ml (i.e. dissolve 2.4g poly-HEMA in 20ml 95% ethanol), shake and mix at 65°C for 8 hours to obtain poly-HEMA HEMA stock solution; then dilute the poly-HEMA stock solution with 95% ethanol at a volume ratio of 1:10 to make a poly-HEMA working solution with a final concentration of 12 mg / ml, store it at 4°C, and set aside; take poly-HEMA Spread the HEMA working solution evenly on the bo...

Embodiment 2

[0056] Example 2 Dynamic Simulation of Reaching Ectopic Reattachment Growth During Cell Transfer

[0057] Metastatic attachment-resistant hepatoma cells mimicking survival in the vasculature for 24 h were harvested and repopulated into poly-HEMA-free cell culture plates at 37 °C in 5% CO 2 Continue culturing in the incubator for 24h-48h, and record the morphological data with a Nikon8700 camera every 12 hours. At the same time, the cells that are about to adhere to the wall again after the suspension are collected at 12 hours and 24 hours of culture, and processed into a single cell suspension, washed with PBS, fixed overnight with 70% ethanol, propidium bromide (PI, Sigma product) Staining, flow cytometry to detect the distribution of cells in each cell cycle.

[0058]Collect cells suspended in 96-well plates for 24 hours, transfer them into 96-well plates without poly-HEMA, add 10ul CCK8 to each group of 3 duplicate wells every 12 hours, mix well, and place at 37°C , CO 2...

Embodiment 3

[0060] Example 3 Detection of apoptosis rate of liver cancer cells in metastatic state

[0061] A poly-HEMA-coated 6-well cell culture plate was prepared, and the 6-well cell culture plate was divided into a suspension culture group and an adherent culture group. Collect the hepatic carcinoma cells suspended and attached to the wall for 24 and 36 hours respectively, and centrifuge at 1000rpm for 5min; every 10 6 Add 1ml of lysis / sample buffer (0.2mM) to each cell, place on ice for 30min; collect the lysate and precipitate of liver cancer cells suspended and adherent for 24 and 36 hours, and centrifuge at 3000rpm for 10min; store at -20°C for more than 18 hours; Take out the sample precipitate at -20°C, dissolve the precipitate with 10 times the volume of lysis / sample buffer; take the sample buffer and add it to the microplate, 100ul / well, seal it with glue strips, and store it at room temperature for 3 hours; discard the supernatant, add 1× wash Wash with the buffer three tim...

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Abstract

The invention discloses a model in vitro which can simulate the metastasis of human liver cancer dynamically. The construction steps comprise the construction of an anti-cell adherence model by using human liver cancer cell line BEL7402 with high metastasis; the construction of metastasis model in vitro of liver cancer cell and the identification for the constructed in vitro metastasis model of liver cancer. The model of the invention can be used for the control research between the in situ cancer and the tumor in metastasis; the invention can further be used as a monitoring system for the biological behavior detection of the tumor cell in metastasis and the judgment of tumor prognosis. The model has important value for the research of clinical tumor metastasis mechanism and the screening of metastasis control drugs. The model can also be used as the screening technology platform for the preparation of anti-tumor transgenic vaccine and the construction of anti-tumor metastasis drugs.

Description

technical field [0001] The invention relates to a human liver cancer metastasis model, in particular to an in vitro model for dynamically simulating human liver cancer metastasis. Background technique [0002] As a malignant tumor with extremely high morbidity and mortality in China, liver cancer has become a long-term problem for clinicians and scientific researchers because of its rapid disease progression, high degree of malignancy, high metastasis rate, and high recurrence rate. Although a small number of patients have achieved long-term survival due to early diagnosis and treatment, the metastasis and recurrence of liver cancer have always been a major obstacle affecting the treatment effect of liver cancer. Clarifying the mechanism of liver cancer metastasis and its early diagnosis and prevention will be the key to further improve the treatment effect of liver cancer. [0003] Liver cancer is a chronic disease that seriously endangers human health. The treatment metho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12N5/09
Inventor 孙汶生韩丽辉张之勇曹莉莉梁晓红曲忠花杜娟马恬侯楠郭春刘玉刚
Owner SHANDONG UNIV
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