Brucein D Inhibits Proliferation And Induces Apoptosis In Colon Adenocarcinoma Cell Line HT29

Objective To investigate that brucein D (BD) possesses potent anti-colon cancer activity by inhibiting proliferation and inducing apoptosis in colon adenocarcinoma cell.Methods After human colon cancer cell line PANC-1 was treated with BD (0,5,10,20, 30,80)?mol?mol/L for 24?48?72 h, the cell proliferation was detected by cell counting kit-8(CCK-8)assay. The apoptosis of colon cancer cells were detected by using Annexin V-FITC/PI. The protein expressions of JNK and p-JNK were detected by using Western blot. HT29 cells were injected subcutaneously into nude mice to establish colon xenograft tumors. Tumor bearing animals were divided into the following treatment groups:control group,0? 0.375mg/Kg·d group?0.75 mg/Kg·d group,1.5 mg/Kg·d group. Each group consisted of ten nude mice. After the mice were treated for 2 weeks, the volumes of xenograft tumors were monitored. Mice were sacrificed by cervical dislocation after the next 2 weeks, Western blotting was used to detect the protein expression of JNK and p-JNK in colon tumor.Results 1. After human colon cancer cell line PANC-1 was treated with BD (0?5?10? 20?30?80?mol/L) for 24 h, the cell survival rates, detected by CCK-8, were 100%?93.83%? 82.50%?71.16%?63.39%?38.56%.For 24 h, the cell survival rates were 100%?82.87%? 68.62%?52.25%?39.33%?28.55%, For 48 h, the cell survival rates were 100%?72.60%? 55.37%?40.68%?34.68%?20.01%.2. After human colon cancer cell line PANC-1 was treated with BD (0?5?10?20?30?80?mol /L) for 24 h, detected by Flow Cytometry, HT29 cells apoptosis rates were 0?10.32%? 19.36%?30.61%?43.47%?66.56%. For 48 h, cells apoptosis rates were 0?19.23%?31.26%? 47.63%?58.91%?70.08%. For 72 h, cells apoptosis rates were 0?27.64%?45.22%?58.28%? 64.56%?79.37%.3. AS the concentration of BD got higher or the time got longer, the protein expression of JNK was up-regulated and that of p-JNK down-regulated significantly.4. In vivo, with the administration prolonged, the mean volume of xenograft tumor growed slowly.5. The protein expression of JNK was up-regulated and that of p-JNK down-regulated significantly in colon xenograft tumors cells.Conclusions In vivo, BD significantly inhibits the proliferation of colon cells in concentration-and time-dependent manners. The higher the concentration of BD, the much stronger the inhibition is. With the administration prolonged, BD's inhibition on the colon cells becomes more obvious. AS the concentration of BD gets higher or the time gets longer, the protein expression of JNK is up-regulated and that of p-JNK down-regulated significantly. In vivo, with the administration prolonged, the mean volume of xenograft tumor grows slowly. The protein expression of JNK is up-regulated and that of p-JNK down-regulated significantly in colon xenograft tumors cells.