| Objective:In this study, we investigate the expression and significance of Weel and its phosphated forms, and analyze the effect of Weel inhibitor PD 407824 on human colorectal cancer cell proliferation. We aim not only to investigate the the molecular mechanism of tunlor proliferation, but also to search new effective target for gene therapy of colorectal cancer.Methods:1. At first, western blotting analysis was used to detect the expressions of Weel different colorectal cancer cells, including lovo,HCT116,SW620,SW480。Then we chose the one who expresses more to continue the following research.2. After the starvation to make the cells synchronization, we used western blotting analysis to detect the expressions of the Wee1,p-WeelSer53 and p-WeelSer642 in the colorectal cancer cell.3. The cells were cultured in medium containing different concentrations of Weel inhibitor PD 407824. Cell proliferation was determined using Cell Counting Kit-8(CCK-8) solution, and the shapes of the cells were observed with the microscopy.Results:1. HCT116 cell expressed more Weel in the four different cells (lovo,HCT116,SW620,SW480) 2. At the different time, the expressions of the Weel and its phosphorylated forms were rages. They were rising for 6h,12h,24h after releasing from starvation, and the time with highest expression for Wee1,p-WeelSer53 and p-WeelSer642 is 6h,12h and 12h, respectively.3. Treated with different concentrations of Weel inhibitor PD 407824, the colorectal cell proliferation was inhibited.4. Using microscopy, we observed that the number of HCT116 cells was decreased and the cells turned small and round with different concentrations different concentrations of Weel inhibitor PD 407824.Conclusion:1. The activity and degradation of Weel may closely relate to the sites and levels of phosphorylation.2. Weel inhibitor can inhibited the cells proliferation which have more expression of Weel.3. Weel inhibitor can considered as a new treatment of colorectal tumors in the future, hopefully. |
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