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目的探讨受体作用蛋白(receptor-interaction proteins,RIP)在异硫氰酸苯乙酯(phenethy lisothiocyanate,PEITC)诱导人白血病细胞凋亡中的作用。方法将0、2、4、6、8μmol/L异硫氰酸苯乙酯作用于U937细胞3、6h,观察量效关系。用8μmol/L异硫氰酸苯乙酯作用U937细胞1、3、6、9、12、24h,观察时效关系。采用AnnexinV/PI染色和流式细胞仪检测细胞凋亡。用Westernblot法检测细胞凋亡相关蛋白蛋白聚ADP核糖聚合酶[poly(ADP-ribose)polymerase,PARP]、Caspasep-3、Caspasep-8及RIP的表达。结果异硫氰酸苯乙酯在2、4、6、8μmol/L作用于U937细胞3h分别引起9%、25%、44%和56%的细胞发生凋亡,6h分别引起16%、28%、51%和82%的细胞发生凋亡,呈明显的量效关系。8μmol/L的异硫氰酸苯乙酯作用U937细胞1、3、6、9、12、24h分别引起12%、57%、79%、86%、90%和91%的细胞发生凋亡,呈明显的时效关系。Westernblot结果表明,异硫氰酸苯乙酯引起凋亡蛋白酶Caspase-3、Caspase-8的降解/活化增加,促进凋亡作用底物PARP的降解增加。异硫氰酸苯乙酯还可引起受体作用蛋白表达降低及RIP降解增加。结论异硫氰酸苯乙酯可引起人白血病细胞发生凋亡,其分子机制可能与死亡受体途径有关,其中受体作用蛋白的降解可能在异硫氰酸苯乙酯诱导细胞凋亡中起着重要作用。
Objective To investigate the role of receptor-interaction proteins (RIP) in the apoptosis of human leukemia cells induced by phenethylisothiocyanate (PEITC). Methods Phenyisyl isothiocyanate (0,2,4,6,8μmol / L) was applied to U937 cells for 3 and 6 hours, and the dose-response relationship was observed. U937 cells were treated with 8μmol / L phenethyl isothiocyanate for 1, 3, 6, 9, 12 and 24 hours. AnnexinV / PI staining and flow cytometry were used to detect apoptosis. The expressions of PARP, Caspasep-3, Caspasep-8 and RIP were detected by Western blot. Results Phenylallyl isothiocyanate induced cell apoptosis in U937 cells treated with 2,4,6,8μmol / L for 3h, respectively, at 9%, 25%, 44% and 56% , 51% and 82% of cells apoptosis, showed a significant dose-response relationship. The cells treated with 8μmol / L phenethyl isothiocyanate for 1, 3, 6, 9, 12 and 24 h induced apoptosis in 12%, 57%, 79%, 86%, 90% and 91% Obvious aging relationship. Western blot results showed that PES induced the degradation / activation of Caspase-3 and Caspase-8, and promoted the degradation of PARP, an inhibitor of apoptosis. Phenylethyl isothiocyanate also causes a decrease in receptor protein expression and an increase in RIP degradation. Conclusion Phenethyl isothiocyanate can induce apoptosis in human leukemia cells, and its molecular mechanism may be related to the death receptor pathway. The degradation of receptor-acting proteins may play a role in the induction of apoptosis by phenylethyl isothiocyanate An important role.