Abstract: Background and purpose: Aspirin (one of the nonsteroidal anti-inflammatory drugs) is effective in the prevention and treatment of colon cancer. And recently, it has been indicated that regulating colon cancer stem cells might be the underlying mechanism. When colon cancer stem cells become mature, colon cancer will not progress, and even subside. Therefore, this study aimed to explore the effects of aspirin on differentiation in human colon cancer cell line HT-29. Methods: The inhibitory effects of different concentrations of aspirin on HT-29 cells were detected by living cell counting kit CCK-8, and the IC₅₀ was calculated. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) was used to detect the mRNA expressions of intestinal differentiation markers [mucin 2 (MUC2), trefoil factor 3 (TFF3) lysozyme and sucrase-isomaltase] after aspirin IC₅₀ intervention for 24, 48 and 72 h. Immunocytochemistry was used to detect the MUC2 protein expression. And Western blot was used to detect the protein expressions of lysozyme and sucrase-isomaltase. Results: Aspirin inhibited the proliferation of HT-29 cells significantly in a time- and dose-dependent manner. RTFQ-PCR and Western blot showed that the mRNA expressions of goblet cell markers (MUC2 and TFF3) were increased (P<0.05), and the mRNA and protein expressions of Paneth cell marker (lysozyme) and absorptive cell marker (sucrase-isomaltase) were decreased (P<0.05), compared with the control group, after aspirin intervention for 48 and 72 h. Immunocytochemistry showed that the MUC2 protein expression was increased (P<0.05) compared with the control group after aspirin intervention for 48 h. Conclusion: Nonsteroidal anti-inflammatory drug aspirin can affect the expressions of intestinal differentiation markers in human colon cancer HT-29 cells, and may lead to their differentiation into the phenotype of goblet cell.

Key words: Aspirin, Colon cancer, Mucin 2, Trefoil factor 3, Sucrase-isomaltase, Lysozyme