Objective To observe the effect of stable cyclin-dependent kinase 2(CDK2)siRNA transfection on proteomic expression in human hepatoma HepG2 cells,and to investigate the role of CDK2 in the pathogenesis of hepatocarcinoma.Methods Two-dimensional gel electrophoresis-mass spectrometry(2-DE-MS)was used to detect the protein expression in CDK2shRNA-trans-fected HepG2 cells(HepG2-CDK2shRNA cells)and HepG2 cells.Results Ten differential ex-pression proteins were identified by 2-DE-MS.Of them,7 proteins(cytokeratin 18,chaperonin 10-related proteins(HSP10,HSPE1 and CPNl0),human chorionic gonadotropin,Kv channel interac-ting protein 3,olfactory receptors(family 2,subfamily L and member 2),chromosome 9 open read-ing frame 3,and ELYS transcription factor-like protein TMBS62)were not expressed in HepG2-CDK2shRNA cells,and 3 proteins(high-mobility group box 1,heterogeneous nuclear ribonucleo-protein H1 and immunoglobulin heavy chain variable region)were down-regulated in HepG2-CDK2shRNA cells.Conclusion There were significant changes in expressed proteins involved in cell cycle,signal transduction,and tumor metastasis and infiltration after the inhibition of CDK2 expression in human hepatoma HepG2 cells.%目的:观察稳定转染细胞周期蛋白依赖性激酶2(cyclin-dependent kinase 2,CDK2)干扰 RNA 对人肝癌细胞株 HepG2蛋白质组表达的变化,探讨 CDK2在肝癌发生中的作用。方法采用双向凝胶电泳-质谱技术比较稳定转染 CDK2干扰 RNA 的 HepG2-CDK2shRNA 细胞株及 HepG2细胞株蛋白质表达的变化。结果经双向电泳-图像分析-质谱技术得到10个差异表达的蛋白质。其中7种[细胞角蛋白18((cytokeratin18,CK18))、热休克蛋白家族成员伴侣素10(chaperonin 10-related protein,Hsp10、HSPE1、CPNl0)、人绒毛膜促性腺激素(hCG2039058)、钾离子通道蛋白3(Kv channel interacting protein 3,Kv3)、嗅觉受体(olfactory receptor,family 2、subfamily L、member 2)、染色体9开放性读码框架3(chromosome 9 open reading frame 3)、ELYS 转录因子样蛋白(ELYS transcription factor-like protein TMBS62)]在 HepG2-CDK2shRNA 细胞中不表达,3种[高迁移率族蛋白1(high-mobility group box 1,HMGB1)、异源核糖核蛋白 H1(heterogeneous nuclear ribonucleoprotein H1,hnRNP H1)、免疫球蛋白重链可变区(immunoglobulin heavy chain variable region,IgVH)]表达下调。结论CDK2表达抑制后人肝癌细胞 HepG2在细胞周期、信号转导、肿瘤转移和浸润等方面的蛋白质表达存在显著差异。
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