Human breast cancer cell line with high lung metastasis

A high-metastasis technology of human breast cancer cells, applied in the field of microbial animal cell lines, can solve the problems of short incubation period and low rate of spontaneous lung metastasis, and achieve the effects of wide application, short experimental period, and clear genetic background

Inactive Publication Date: 2010-06-23
FUDAN UNIV SHANGHAI CANCER CENT
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Problems solved by technology

However, in recent years, there has been considerable controversy about the cell origin of MDA-MB-435. Some studies have pointed out that this cell line may come from melanoma rather than breast cancer (Nat Genet, 2000); MDA-MB-231 is also a commonly used human breast cancer cell line. Cancer cell line has the advantages of high tumor formation rate (100%) and short incubation period (15 days), but its spontaneous lung metastasis rate is not high, generally not more than 10%

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  • Human breast cancer cell line with high lung metastasis
  • Human breast cancer cell line with high lung metastasis

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Embodiment 1

[0014] Example 1 Screening and establishment of lung metastatic cells

[0015] The cells involved in the present invention are all cultured in Leibovitz's L-15 cell culture medium containing 10% fetal calf serum (FCS). The culture temperature was 37°C, and the gas environment was 5% CO 2 / 95% air, humidity is saturated humidity. When subcultured, the cells were digested with 0.25% trypsin, and subcultured at a ratio of 1:3.

[0016] 1. The first round of screening of lung metastatic cells

[0017] ① Digest MDA-MB-231 cells in the logarithmic growth phase with 0.25% trypsin to make a single cell suspension;

[0018] ②Under sterile conditions, tumor cells were inoculated into the fat pad (mfp) of the left second papilla of 6-week-old BALB / c female nude mice (BALB / c nude mice) with a 1ml syringe or directly injected into the tail vein (i.v. ), the capacity is adjusted to 1×10 7 Cells / 0.1ml / one (mfp) or 2×10 6 Cells / 0.1ml / piece (i.v.);

[0019] ③After 4 weeks of routine fee...

Embodiment 2

[0031] Example 2 Mice Transplanted Tumor and Lung Metastasis Experiment

[0032] 1. BALB / c nude mice

[0033] ①Digest the MDA-MB-231 and MDA-MB-231 / 08-001H cells in the logarithmic growth phase with 0.25% trypsin, and make single-cell suspensions respectively;

[0034] ②Under sterile conditions, use a 1ml syringe to inoculate tumor cells into the fat pad of the left second papilla of nude mice, with an inoculation volume of 1×10 7 Cells / 0.1ml / piece;

[0035] ③Observe the health status of the nude mice and the tumorigenicity of the tumor cells the next day;

[0036] ④After 4 weeks, 6 nude mice were randomly selected from each group, and their lungs were fixed with Bouin's solution for 24 hours, and then soaked in absolute alcohol until the color of the lung tissue recovered and the metastatic lesions were white nodules, and were examined under a dissecting microscope. The number of metastatic nodules in each lung was observed and recorded; some transplanted tumors were excis...

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Abstract

The invention belongs to the field of microbial and animal cell lines, and particularly relates to a human breast cancer cell line MDA-MB-231 / 08-001 with spontaneous high lung metastasis potentiality and an establishment method thereof. The cells of the cell line are humanized cells that have 92 expression genes different from those of parent cells, higher in-vitro growth capacity, multiplication capacity, anti-invasion capacity, tumor forming property and lung metastasis than the parent cells; and in 4 weeks after the cells are inoculated, the in-situ tumor volume of a scid mouse is 4,224.67+ / -1,038.95mm<3>, the weight is 2.09+ / -0.51g, the spontaneous lung metastasis is 100 percent, and the number in a metastasis lesion is 41 in each lung. The cell line and an established animal model can simulate the whole process from the growth of the tumor cells at implantation position to the formation of the metastasis lesion at a target organ, which is quite consistent with the clinical practical condition, and can be conveniently used for the research of breast cancer lung metastasis mechanisms and related medicaments.

Description

technical field [0001] The invention belongs to the field of microbial animal cell lines, and in particular relates to a human breast cancer cell line with spontaneous lung high metastatic potential and a method for establishing the same. Background technique [0002] According to reports, breast cancer has become the primary malignant tumor that seriously endangers women's health. According to the 2008 cancer statistics report, if American women live to the age of 85, 1 / 8 of them (about 12.7%) will suffer from breast cancer in their lifetime. Breast cancer ranks first among all female tumors, accounting for 26%, far surpassing the second-ranked lung cancer and the third-ranked colorectal cancer, leaving other cancers behind. In recent years, with the development of the economy and the change of people's life style, the incidence of breast cancer in our country has also shown an obvious upward trend, ranking first among all kinds of malignant tumors among women in big citie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08
Inventor 邵志敏欧周罗侯意枫罗建民
Owner FUDAN UNIV SHANGHAI CANCER CENT
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