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Archive > Volume 23 Issue 4 > 2016,23(4):486-491. DOI:10.3872/j.issn.1007-385X.2016.04.005 Prev Next
Basic Research
Establishment of a gastric cancer cell line with high metastasis and biological characteristics of cancer stem cell of the cell line
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Abstract:
Objective:To establish a gastric cancer cell line with high metastasis and explore biological characteristics of cancer stem cell for the cell line. Methods: The gastric cancer SNU-5 line cells were subcutaneously injected into nude mice. After a xenotransplanted tumor was formed, pulmonary metastasis of the tumor was taken out and repeatedly injected into nude mice by mechanical separation method, as well as the SNU-5-V12 cell line was achieved. Serum free suspension culture and PKH26 staining were used to confirm that there are cancer stem cells in the SNU-5-V12 cells. Expressions of cancer stem cell marker CD44 in the SUN-5 and the SUN-5-V12 cell lines were analyzed with flow cytometry assay. CD44+ cells in the SNU-5 and the SNU-5-V12 cell lines were separated and research in vitro of biological characteristics and tumorigenic experiments in nude mice were performed with the CD44+ cells. Results:The SNU-5-V12 cell line with high metastasis was established. There was a single PKH26 positive cell in spheroids formed by the SNU-5-V12 cells after serum free suspension culture for 11 days. Analysis of flow cytometry shown that ratio of cancer stem cell marker CD44 in SNU-5-V12 cell with high metastasis was significantly higher than that in the SNU-5 cell (\[72.9±1.5\]% vs \[8.96±1.2\]%), and self-renewal capacity of the CD44+ cells in the SNU-5-V12 cells was obviously stronger than that in the SNU-5 cells (forming spheroid rate: \[27.8±1.7\]% vs \[20.4±10\]%, P<0.01), as well as invasion ability of the SNU-5-V12 cells increased 1.6 times (\[329.5±7.5\] vs \[200±20\], P<0.01）, its drug resistant ability also enhanced significantly (IC50: \[0.286 vs 0.196\]μg/ml, P<0.01). At two months of the injection, third one nude mice (2/6) that injected with 2×102 the SNU-5-V12 cells formed xenotransplanted tumor, but only minority nude mice (1/6) that injected with 2×104 the SNU-5 cells formed xenotransplanted tumor. Conclusion:A gastric cancer the SNU-5-V12 cell line with high metastasis was obtained. In vivo and in vitro functions of CD44+ cell in the SNU-5-V12 cell line significantly increased. It could provide a valuable cell model for target therapy of gastric cancer stem cell.
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Project Supported:
Project supported by the National Natural Science Foundation of China（No. 31371445）, and the National High-tech R & D Program of China (863 Program) (No. 2014AA020537)
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