Human gastric cancer cell line with 5-fluorouracil resistance and establishing method and application thereof
A gastric cancer cell line and a method for establishing a technology, applied in the biological field, can solve the problems of a large difference in the biological characteristics of gastric cancer and low biodiversity of human gastric cancer cell lines, and achieve high scientific research value and development significance, high homogeneity, and success. The effect of high tumor rate
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Embodiment 1
[0048] The preparation of embodiment 1GAXC066 cell
[0049] Fresh clinical gastric cancer resection specimens were obtained from Nantong Cancer Hospital (patient sex: male, age: 72 years old, ethnicity: Han, native place: Nantong City, Jiangsu Province.), and the clinical diagnosis was poorly differentiated gastric adenocarcinoma.
[0050] Rinse with fresh HBSS buffer (containing 500 U / ml penicillin G, 500 μg / ml streptomycin sulfate and 1.25 μg / ml amphotericin B), cut into small pieces of 20-50 mg, subcutaneously puncture and inoculate in immunodeficient animals SCID mice (SCID mice were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.). 70-90 days after puncture inoculation, the tumor-bearing animals were sacrificed, and the tumor tissues were taken out for primary culture and subculture of cancer cells.
[0051] The primary culture steps are as follows: cut the tumor tissue into small pieces, place them in a culture bottle, and incubate at 37°C ...
Embodiment 2
[0054] Biological characteristics and application of embodiment 2GAXC066 cells
[0055] In the present invention, RPMI1640 is used for culturing and the GAXC066 cells are purified by a differential attachment method, so that they can grow long-term in vitro and pass stably. When the cells are passed to more than 20 generations, the cell properties are gradually stabilized, and the relevant biology, genetics and tissue source identification are carried out until the 50th generation has the same stable properties. Through experimental observation and verification, GAXC066 main cells grown in vitro are in the shape of large willow leaves and have no contact inhibition properties. Genetic studies have confirmed that the cells are aneuploid, with severe chromosome number and structural aberrations, consistent with the genetic characteristics of malignant tumors. The cells can form tumors in nude mice and have tumorigenicity. The corresponding relationship between the cells and th...
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