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The Effect of HDACs Inhibitor SAHA on p21 Protein Stability in U251 Cells
Gong Aihua1,2*, Xiong Ermeng1, Zhang Yan1, Du Fengyi1, Peng Wanxin1, Shao Genbao1, Jin Jie1, Cheng Jianjun1
1School of Medical Science and Laboratory Science, Jiangsu University, Zhenjiang 212013, China;2State Key Laboratory for Oncogenes and Related Genes, Shanghai 200032, China
Abstract: Previous studies suggest that HDACs inhibitors including SAHA are proming drugs against tumors in clinical trails. However， the mechanism of SAHA against tumor cells still remains to be clarified. In this study， we investigated the effects of SAHA on the expression of p21 mRNA and protein in tumor cells using Realtime PCR and Western blot， and the stability and ubiquitination of p21 protein mediated by GSK-3β in U251MG cells through Western blot and Co-IP assay. Subsequently， cell cycle progress was determinded using FACS in U251MG cells treated with SAHA and transfected with vector， GSK-3βKD or GSK-3βCA. We found that SAHA upregulated both p21 mRNA and protein levels， and decreased ubquitination levels of p21 protein and thus enhanced its stability， and arrested cell cycle progress at G1 phase. Our finding confirmed that SAHA enhanced the stability of p21 protein via inhibition of GSK-3β activity， and thus played roles in blocking cell cycle progress，which might provide the evidence for further clinical research against glioma cells.
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