| Abstract: | AIM To explore the anti-tumor effect of indomethacin (IN) on human colon adenocarcinoma cells anddetermine the influence of indomethacin on cancer cell proliferation and apoptosis and elucidate the anti-tumor mechanism of Indomethacin.METHODS Human colon adenocarcinoma HCT116 cell line were cultured separately in vitro. Indomethacin(final concentration 100 μm - 800 gm) was administered alone or altogether with 5-Fu (50 μm). Agarose gelelectrophoresis, MTT, and Flow cytometry were used to study cell proliferation and apoptosis in humancolon carcinoma cell RT-PCR, western blot were used to detect the expression level of Bcl-2, bax gene andcdk4 protein expression in HCT116 cell lines after treated with IN for 24 hours.RESULTS Indomethacin can inhibit significantly the proliferation of HCT116 cell, change the morphology,and cause the cells to accumulate in the G0/Gl phase of the cell cycle, and induce apoptosis. The apoptosis oftumor cells was confirmed by DNA ladder formation on gel electrophoresis and sub-Gl peak on flowcytometry. These responses were time-and concentration-dependent. A synergic effect of inhibiting cancercell proliferation was observed when combined with Indomethacin and 5-Fu. RT-PCR results showed that INdown-regulated Bcl-2 mRNA expression, and did not change Bax mRNA expression. Western blot resultsconfirmed that IN inhibited Bcl-2 protein expression. No influence was found in the translation of Baxprotein. In inhibited cdk4 protein expression.CONCLUSION Our study results indicate that IN induce apoptosis of HCT116 cell by down-regulating Bcl-2expression and inhibiting cdk4 protein expression partially. This explains the mechanisms of antitumoractivity of the Indomethacin. |
