目的观察HepG2细胞中NFκBp65基因沉默对FAT10蛋白表达的影响。方法采用电穿孔法将p65siRNA转染人肝癌细胞系HepG2细胞,采用Western blot检测各组细胞中p65和FAT10蛋白的表达变化,用染色体核型分析p65干扰对FAT10过表达而导致的非整倍体现象的阻断作用。结果Western blot结果显示,p65 siRNA干扰可以有效抑制HepG2细胞中NFKBp65的表达,并且通过抑制p65可以有效抑制TNF-α诱导的FAT10高表达,进而通过有效抑制TNF-α诱导的FAT10高表达而阻断TNF-α诱导的非整倍体现象。结论NFKBp65的表达下调可以抑制TNF-α诱导的FAT10高表达及非整倍体现象。
Objective To investigate the FAT10 protein expression in HepG2 cells after NFκB p65 gene silence induced by siRNA. Methods Electroporation system was used to transfect siRNA into HepG2 cells. Western blot was employed to detect the expression of FAT10 and p65 proteins in HepG2 cells after transfection. Cytogenetic analysis was performed to analyze the chromosome karyotype. Results NFκB p65 gene expression was effectively inhibited by siRNA. The TNF-α induced over-expression of FATIO gene was significantly impaired after p65 siRNA transfection. At the same time, TNF-α induced abnormal chromosome karyotype were also blocked. Conclusion siRNA in- terference targeting p65 can inhibit the expression of FATIO gene and TNF-α induced abnormal chromosome karyotype in HepG2 ceils.