目的研究CD59配体肽基因对卵巢癌细胞CD59表达的影响。方法构建CD59配体肽基因的真核表达重组体(CD59配体-pIRES),转染人卵巢癌细胞A2780,G418筛选稳定表达细胞克隆;RT-PCR及Westernblot方法检测细胞表面CD59 mRNA及蛋白的表达;MTT法测定细胞增殖抑制率。结果成功构建了CD59配体肽基因真核表达重组体,转染后的A2780 CD59 mRNA及蛋白的表达水平与WT-A2780比较均明显下降(t=9.217、.20,P〈0.05);细胞增殖抑制率明显增高(F=5.417,q=3.93,P〈0.05)。结论CD59配体肽基因可影响人卵巢癌细胞A2780表面CD59的表达,有望用于肿瘤治疗。
Objective To study the influence of CD59 ligand-peptide on ovarian cancer cell CD59. Methods A eukaryotic expression recombinant of ligand-peptide of CD59 was created and transfected into A2780 cells.Stable expressive cell clone was screened by addition of G418.The level of CD59 mRNA and protein was detected by RT-PCR;and cytostasis rate detected by MTT. Results A eukaryotic expression recombinant of ligand-peptide of CD59 was successfully constructed.The expressions of CD59 mRNA and protein in transfected A2780 cells were lower than that of WT-A2780(t=9.21,7.20;P0.05),and cytostasis rate obviously increased(F=5.417,q=3.93,P0.05). Conclusion CD59 Ligand-peptide can influence the expression of CD59 mRNA and protein on the surface of A2780 cells,which is hope to be used for oncotherapy.