【摘要】 目的初步研究Wnt通路及蛋白激酶B(PKB)在非甾体类抗炎药sulindac诱导人肝癌SMMC-7721细胞凋亡过程的作用。方法利用流式细胞仪检测肝癌细胞7721的凋亡,利用Western blot检测caspase-9、PARP(poly ADP-ribose polymerase)等凋亡相关分子的剪切,β连环蛋白(β-catenin)及糖原合酶激酶3β(GSK3β)与蛋白激酶B(PKB)磷酸化水平的变化,利用RT-PCR检测β-catenin及c-myc的mRNA水平。结果sulindac能够诱导SMMC-7721细胞凋亡,显著降低β-catenin的蛋白水平,抑制GSK3β9位丝氨酸的磷酸化,同时促使caspase-9、PARP等凋亡相关分子发生剪切;β-catenin的mRNA水平未见变化;PKB的磷酸化水平未见降低,反而略有增高。结论sulindac能够通过抑制Wnt通路,促进β-catenin的蛋白降解,降低其蛋白水平,诱导SMMC-7721细胞凋亡,且该作用不依赖PKB活性的抑制。更多还原

【Abstract】 Purpose To study the role of Wnt pathway and protein kinase B(PKB) in sulindac,one of nonsteroidal anti-inflammatory drugs,induced human hepatocellular carcinoma SMMC-7721 cell apoptosis. Methods The apoptosis of hepatocellular carcinoma SMMC-7721 cells were detected by flow cytometry.Caspase-9,PARP(poly ADP-ribose polymerase),β-catenin,GSK3β and protein kinase B(PKB) were detected by Western blot.The mRNA level of β-catenin and c-myc were detected by PT-PCR. Results Sulindac could cause SMMC-7721 cells death through cleavage of caspase-9 and PARP,and decline the protein level of β-catenin but not the mRNA level.Also the phosphorylation level of GSK3β was inhibited.However,the phosphorylation level of PKB did not decline but slightly enhanced. Conclusions Sulindac induced SMMC-7721 cells apoptosis through inhibiting Wnt pathway by declining the protein level of β-catenin,which was independent of PKB inhibition.更多还原