| Objective: Based on the preliminary identification of side population(SP) isolated from cell lines of gastric cancer, to investigate the feasibility of starting research on gastric cancer stem cell from the side population.Methods: Three human gastric cancer cell lines with different differentiation grades, named MKN-28, SGC-7901, and BGC-823, were stained by fluorochrome Hoechst33342 and PI, controlled with the presence of verapamil, and then the SP cells were analyzed and isolated by fluorescence-activated cell sorting( FACS ). Cell growth curve( CCK-8 ) and colony formation assay were used to identify the biological characteristics of the SP cells isolated from Cell line MKN-28.Results: (1) Side population were detected in all of the three gastric cancer cell lines, with a proportion of 0.8%-2.7%. (2) Proportion of the SP was related to the differentiation grade of the cell line, and more poorly the cell line differentiated, the fewer proportion it contained ( P= 0.023). (3) Cell growth curve indicated that proliferative capacity of the side population isolated from MKN-28 was better than the total cells without sorting and the non-SP cells( P= 0.044). (4) Colony formation assay showed that the colony-forming efficiency of the side population was higher than the total cells without sorting and the non-SP cell(89.33%±6.11% vs 42.67%±6.43% vs 13.33%±1.16%, P= 0.026) (5) Through 2 weeks of culture, SP cells isolated from MKN-28 could finally generate a group of cells with a SP proportion of 1.4% , while the non-SP cells isolated synchronously generated a group of SP cells with a fewer proportion of 0.1 %.Conclusion: Side population could be detected in the cell lines of gastric cancer, and it had a fewer proportion in the more poorly differentiated ones. The SP, with the ability of dissymmetry fission, has better proliferative capacity in vitro and higher colony-forming efficiency than the non-SP. So it can be applied to the gastric cancer stem cell research. |
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