Study on the proliferation mechanism of colorectal cancer cells in three-dimensio and two-dimensio culture environment
Objective To investigate the change and mechanism of proliferation ability of colorectal cancer cells in three-dimensio(3D)and two-dimensio(2D)culture environment.Methods SW480 colorectal cancer cells were purchased from Shanghai Cell Bank.SW480 colorectal cancer cells were cultured by chitosan-collagen hydrogel 3D and cell culture dish 2D respectively.The cell morphology was observed by optical microscope.Cell proliferation was detected with cell counting kit-8(CCK-8)cell counting kit.The expressions of OTUB2 and PCNA in 3D and 2D culture were detected by Western blot.The expression of OTUB2,PCNA gene and pro-tein and cell proliferation rate of SW480 cells before and after transfection were compared.Results SW480 colon cancer cells could stick to the wall after 24 hours in 2D environment,and the attached cells slowly spread out and showed short rod shape after 7 days.SW480 colon cancer cells grew in 3D spheres after 3D culture of chitosan-collagen hydrogel for 24 hours,and the number of cells in-creased and the cell spheres became larger after 7 days of culture.The proliferation rate of SW480 cells in 3D culture environment was lower than that in 2D culture environment,and the difference was statistically significant(P<0.05).Western blot analysis showed that compared with 2D culture,the expression of OTUB2 and PCNA protein in colorectal cancer cell lines cultured in 3D culture environ-ment was lower than that in 2D culture environment,and the differences were statistically significant(P<0.05).The expression of OTUB2,PCNA gene and protein and cell proliferation rate of SW480 cells after transfection were higher than those before transfection,with statistical significance(P<0.05).Conclusion The proliferation rate of colorectal cancer cells slowed down under the condition of chitosan-collagen hydrogel 3D culture,and the mechanism is related to the regulation of OTUB2 and PCNA expression.
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NSTL
万方数据
维普
