作　　者： ; ; ; ; ; ;

机构地区： 广州市第一人民医院

出　　处： 《分子影像学杂志》 2019年第3期393-396,共4页

摘　　要： 目的探讨CD74和RUNX3在人胰腺癌Capan-2细胞内是否存在相互作用。方法构建CD74-siRNA的慢病毒表达载体和空白载体,酶切鉴定正确后,分别转染人胰腺癌Capan-2细胞,使用Westernblot和RT-PCR等方法对CD74的表达进行检测验证;运用Westernblot和RT-PCR等方法观察CD74低表达对RUNX3蛋白和mRNA表达的影响;利用免疫共沉淀法验证Capan-2细胞内CD74与RUNX3之间的相互作用。结果CD74在实验组Capan-2细胞中稳定低表达,实验组CD74蛋白(0.09±0.01vs2.78±0.04)、CD74mRNA(0.39±0.001vs1.00±0.00)低于对照组(P<0.05);抑制CD74可降低RUNX3蛋白和mRNA表达水平(P<0.05);CD74抗体免疫共沉淀下来的蛋白复合物中检测到RUNX3,同时RUNX3抗体免疫共沉淀下来的蛋白复合物中也检测到CD74。结论CD74和RUNX3在人胰腺癌Capan-2细胞内存在相互作用。 Objective To investigate the interaction between CD74 and RUNX3 in human pancreatic cancer capan-2 cells. Methods The lentiviral CD74-siRNA vector and blank vector were constructed. Restriction enzyme digestion was constructed correctly. Then Capan-2 cells were transfected by the lentivirus vector. The cell line stably low expression of CD74 was confirmed by Western blot and RT-PCR. The effect of low CD74 expression on RUNX3 protein and mRNA expression were detected by Western blot and RT-PCR. Co-immunoprecipitation assay was performed to verify the interaction between CD74 and RUNX3 in capan-2 cells. Results Protein and mRNA of CD74 were stably and lowly expressed in Capan-2 cells of the experimental group (P<0.05). Decreased CD74 level significantly inhibited RUNX3 protein and mRNA expression (P<0.05). RUNX3 was detected in the protein complex from the immunoprecipitation by using anti-CD74 antibody. CD74 in the protein complex from the immunoprecipitation was detected by using anti-RUNX3 antibody. Conclusion There is an interaction between CD74 and RUNX3 in human pancreatic cancer Capan-2 cells.

关 键 词： 免疫共沉淀 蛋白相互作用

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