| Objective:To investigate the effects of aspirin on CD133,CD44 and Lgr5 which were supposed to be the markers of tumor stem cells, and to provide some mechanisms of aspirin on preventing colonic cancer.Methods:The expressions of CD44, Lgr5 in colon cancer cell line HT-29, SW-480 were observed by laser confocal immunofluorescence double staining. Colorectal cancer cell line HT-29 and SW480 were treated with aspirin at different concentrations (0 mmol/L, 2.5 mmol/L,5.0 mmol/L, 10.Ommol/L respectively, namely: control group, the low concentration group, the middle concentration group and the high concentration group) in vitro. The expressions of CD133 were observed by flow cytometry (FCM). The expressions of CD44 protein and Lgr5 protein in colorectal cancer cell line HT-29 and SW480 treated with aspirin at different concentrations (0mmol/L,2.5mmol/L,5.0mmol/ L,10.0 mmol/L) were detected by Western blot.Result: In the colon cancer cell lines HT-29 and SW480 had exist the Lgr5+ and CD44+cells. Confocal immunofluorescence shows, the expression of CD44 was localized in the plasma membrane, the colour is red. Two cell lines expressing CD44 fluorescence value of the difference was statistically significant (P<0.05). There was a small amount of the expression of Lgr5 was localized in the membrane expression, most of them were expressed in the cytoplasmic, the colour is green. Two cell lines expressing Lgr5 fluorescence was no significant difference (P>0.05). CD44 and Lgr5 in HT-29 and SW480 colon cancer cell lines have a small amount of co-expression part of the yellow. Flow cytometry showed that the proportion of CD133+cells in the HT-29 cell lines were 88.3650±0.997% and in the SW480 cell lines 65.2233±1.523%. After treated with the increasing concentrations of aspirin, the expressions of CD 133 in colon cancer cell line HT-29 and SW480 showed a decreased trend. There were significant difference between the control group and different concentrations groups (P<0.05, respectively). Western blot results showed that with the increase of concentrations of aspirin, CD44 protein expression decreased, strip color from dark to light. There were significant difference between the control group and different concentrations of groups (P<0.05). With the increase of the concentrations of aspirin the expression of Lgr5 protein also showed a decreasing trend. There was significant difference between the control group and different concentrations of groups (P<0.05).Conclusion:1,In the colon cancer cell line HT-29 and SW480 do exist and Lgr5 + and CD44 + cells.2, Aspirin can inhibit the expression of CD133 in the colon cancer cell line HT-29 and SW480, and its inhibitory effects were increased with the increases of the aspirin concentration.3, Aspirin can inhibit the expression of CD44 protein and Lgr5 protein in the colon cancer cell line SW480 andHT-29 and showed a concentration dependent pattern. |
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