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A pair of human stomach cancer cell lines having high metastasis towards livers and lungs and an establishing method thereof

A gastric cancer cell line and high metastasis technology, applied in the field of human gastric cancer cell line and its establishment, can solve the problems of low metastasis rate and long modeling period, and achieve the effects of high degree of malignancy, short experimental period and high organ metastasis rate.

Inactive Publication Date: 2013-10-02
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But even the manual transfer model has a low transfer rate and a long modeling cycle

Method used

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  • A pair of human stomach cancer cell lines having high metastasis towards livers and lungs and an establishing method thereof
  • A pair of human stomach cancer cell lines having high metastasis towards livers and lungs and an establishing method thereof
  • A pair of human stomach cancer cell lines having high metastasis towards livers and lungs and an establishing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Screening and establishment of gastric cancer cell lines with high metastatic potential

[0027] Use gastric cancer cell line for migration experiment (see 1.1 for specific steps), obtain cells that do not penetrate the membrane and cells that penetrate the membrane, and then collect the cells that penetrate the membrane, and then perform the migration experiment. The cells that do not penetrate the membrane and the cells that penetrate the membrane will be separated. Membrane cells, we still only collect the cells that penetrate the membrane, expand the culture, and then perform the migration experiment. Do this 10 times, and repeatedly use the migration experiment to screen the cells that penetrate the membrane each time to obtain a subgroup of cells with strong migration ability. Then use cell subsets with strong migration ability to perform invasion experiments (see 1.2 for specific steps), collect cells that invade and penetrate the membrane each time, and pe...

Embodiment 2

[0032] Example 2. Identification of biological characteristics of gastric cancer cell lines with high metastatic potential

[0033] 2.1 Determination of cell growth curve (MTT experiment)

[0034] A. Inoculate cells: Digest the cells with digestion solution and prepare a single cell suspension with RPMI-1640 medium containing 10% FBS. 3 ~10 4 Cells were seeded in a 96-well culture plate with a volume of 200 μl per well. There were 3 duplicate wells for each cell and 7 96-well plates were seeded; a blank control well was set parallel to the experimental wells without cells and only medium. In the final color comparison, adjust the zero with a blank hole.

[0035] B Cultivate cells: put the culture plate into CO 2 Incubator, at 37℃, 5% CO 2 And under saturated humidity conditions, cultivate for 24 hours;

[0036] C is colored: after 24 hours of culture, add 20μl of MTT solution (5mg / ml) to each well, continue incubating at 37°C for 4 hours, stop the culture, carefully aspirate the cultu...

Embodiment 3

[0056] Example 3 Identification of biological stability of gastric cancer cell lines with high metastatic potential

[0057] Use RPMI-1640 medium containing 10% FBS at 37℃, 5% CO 2 Under saturated humidity conditions, the gastric cancer cell lines with high metastatic potential MKN28-M and SGC7901-M were successively subcultured and cultured for 30 generations, and then tested for their in vitro migration and invasion ability according to the methods of 1.1 and 1.2, and tested their nakedness according to the method of 2.6. Transfer ability in mice.

[0058] The results show that:

[0059] 1. Highly metastatic cell lines isolated from liver and lung metastases

[0060] After the nude mice were injected with tumor cells subcutaneously, they were sacrificed when they were obviously exhausted and were about to die. The liver and lungs were separated. Multiple small gray-white miliary nodules were seen in the liver and lungs. HE staining of tissue sections indicated tumor metastases. Cu...

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Abstract

The invention belongs to the field of microorganism animal cell lines, and especially relates to a pair of human stomach cancer cell lines having high metastasis towards livers and lungs and an establishing method thereof. The human stomach cancer cell lines are characterized in that the human stomach cancer cell lines are MKN28-M and SGC7901-M respectively; the MKN28-M is assigned the accession number CGMCC No.6901; and the SGC7901-M is assigned the accession number CGMCC No.6902. The cell lines are humanized and have no obvious difference from parent cells in in-vitro growth and multiplication capacities, but have obviously enhanced migration, invasion and transfer capabilities over the parent cells. Results of nude mouse tail vein injection visceral metastasis experiments show that the average rate of metastasis towards livers and lungs is 70%. The cell lines have characteristics of high metastasis capacity and high malignancy degree. An animal model established based on the cell lines provides a good platform for further mechanism study of human stomach cancer metastasis. The animal model is an effective and feasible experiment model used for studying mechanisms of human stomach cancer metastasis and evaluating anti-cancer drug curative effects at present.

Description

Technical field [0001] The invention belongs to the field of microbial animal cell lines, and specifically relates to a pair of human gastric cancer cell lines with high liver and lung metastasis tendency and a method for establishing them. Background technique [0002] The incidence and mortality of gastric cancer rank second among malignant tumors, and metastasis is one of the main causes of death in patients with gastric cancer. Gastric cancer metastasis is still a difficult problem faced by people. Whether it is the study of the molecular mechanism of gastric cancer metastasis or the screening of drugs against gastric cancer metastasis, a stable and metastatic cell model is required. [0003] At present, most scholars believe that due to the genetic instability in the development of tumors and the influence of tumor microenvironment on tumor cells, tumor cells gradually appear different phenotypic changes, which is called tumor heterogeneity. That is to say, within the same t...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12R1/91
Inventor 樊代明帖君潘阳林胡凤蓉石晓磊
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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