| Objectives Glycogen synthesis kinases 3(GSK3)β can regulate expansion of breast cancer stem cells, its function is partly dependent on the expression of EZH2 protein. But the molecular mechanisms still remain unclear. Our investigation tried to explain the specific mechanism how GSK3β regulate the expression of EZH2 protein.Methods 1. Examine the correlation between GSK3β activation and EZH2 to BCSC function through introducing control, EZH2, constitutive active GSK3β(GSK3β CA) or kinase dead GSK3β(GSK3β KD) plasmids into breast cancer cells by lentivirus infection. Then calculate the difference ratio of mammosphere counts between the groups. 2. Under the difference conditions of GSK3β activity, using Western blot to detect the expression of EZH2 protein. 3. Using the Co-Immunoprecipition to examine if GSK3β can bind to EZH2. 4. Detect the ubiquitin level of EZH2 Under the difference conditions of GSK3β activity.Results 1. GSK3β activation can affect the expansion of breast cancer cell, its function ispartly dependent on the expression of EZH2. 2. GSK3β can control the expression of EZH2. 3. GSK3β can directly bind with EZH2, increases the EZH2’ ubiquitin level, and decreases the stabilization of EZH2 protein.Conclusion GSK3β can directly bind with EZH2, increases the EZH2’ ubiquitin level, and decreases its stabilization. Its function of regulating the breast cancer cells expansion is partly dependent on the expression of EZH2 protein. |
