Objective: To study-the expression of the serine protease Omi/HtrA2 on apoptosisof hepatoma cells.,Methods: Immunohistochemical technique, real time PCR and Westblot were used to investigate the expression of Omi/HtrA2 in normal hepatocelluar cell line L02, and hepato-celluar carcinoma cell lines; The psiRNA-Omi/HtrA2 was transiently transfected into HepG2 cells and the inhibited effect of psiRNA-Omi/HtrA2 on expression of Omi/HtrA2 was detected using RT-PCR and Western Blot; the apoptosis of cancer cells was determined by flow cytomtry and MTT techniques. Results: 0mi/HtrA2 mRNA and protein expression were detected in all cell lines, but the level of 0mi/HtrA2 mRNA and protein expression in hepatocelluar carcinoma cell lines was higher than that in normal hepatocelluar cell line L02. The apoptosis of PC-3 cells and the expression of Omi/HtrA2 were down-regulated after the transfection of psiRNA-0mi/HtrA2. Conetusiom The expression 0mi/HtrA2 in hepatocelluar carcinoma cell lines was higher, which indicating that O-mi/HtrA2 may play an important role in promoting the apoptosis of hepatocelluar carcinoma and providing a novel option in the aspects of tumor therapy.%目的:探讨Omi/HtrA2丝氨酸蛋白酶活性对肝癌细胞HepG2凋亡的影响.方法:检测Omi/HtrA2在正常肝上皮细胞L02细胞系和肝癌细胞系HepG2、Hep3B、PLC中的表达.用脂质体法将Omi/HtrA2小干扰RNA表达载体(psiRNA-Omi/HtrA2)转染至入HepG2细胞中,以RT-PCR和Western Blot法评价psiRNA-Omi/HtrA2对Omi/HtrA2表达的沉默效应.用MTT法和流式细胞仪检测siRNA导致Omi/HtrA2基因沉默后HepG2细胞凋亡的变化.结果:在L02细胞系和3种肝癌细胞系中,所有细胞系在mRNA水平和蛋白质水平均表达Omi/HtrA2,在肝癌细胞系中的Omi/HtrA2 mRNA和蛋白质表达水平均高于正常肝上皮细胞L02.psiRNA-Omi/HtrA2载体可特异性抑制HepG2细胞中Omi/HtrA2的表达.转染psiRNA-Omi/HtrA2载体的HepG2细胞凋亡下调.结论:Omi/HtrA2在肝癌细胞系中表达增高,它可能在促进肝癌细胞凋亡中有重要作用,Omi/HtrA2为肝癌的治疗提供了一个新的选择靶点.
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