沉默E6相关蛋白对人乳头瘤病毒阴性宫颈癌细胞中p53蛋白表达水平的影响

Effect of silencing E6-associated protein on level of p53 protein in human papilloma virus negative cervical cancer cells

目的探讨沉默E6相关蛋白(E6-associated protein,E6AP)对人乳头瘤病毒(human papilloma virus,HPV)阴性宫颈癌细胞(C33A)中p53蛋白表达水平的影响。方法在LipofectamineTM2000转染试剂的介导下,将特异性沉默E6AP的siRNA序列(siE6AP)及沉默对照无序siRNA序列(siControl)分别转染C33A细胞。Western blot法检测siRNA对E6AP的沉默效果及细胞中p53和cleaved-caspase-3蛋白的表达水平。结果siE6AP组C33A细胞中E6AP蛋白表达水平显著低于siControl组(t=-4.597,P<0.05)。siE6AP组C33A细胞中p53和cleaved-caspase-3蛋白表达水平均显著高于siControl组(t分别为4.533和7.099,P均<0.05)。结论沉默E6AP可显著提高C33A细胞中p53蛋白的表达水平,表明沉默E6AP可能恢复C33A细胞中p53的活性和功能。

Objective To investigate the effect of silencing E6-associated protein(E6AP)on the level of p53 protein in human papilloma virus(HPV)negative cervical cancer cells(C33A cells).Methods The siRNA sequence silencing E6AP(siE6AP)and silencing control disordered siRNA sequence(siControl)were transfected into C33A cells with the mediation of LipofectamineTM2000 transfection reagent respectively.The silencing effect of siRNA on E6AP and the expression of p53and cleaved-caspase-3 proteins were detected by Western blot.Results The levels of E6AP protein in C33A cells of siE6AP group were significantly lower(t=-4.597,P<0.05),while the levels of p53 and cleaved-caspase-3 proteins were significantly higher than those of siControl group(t=4.533 and 7.099 respectively,each P<0.05).Conclusion Silencing of E6AP significantly increased the expression of p53 protein in C33A cells,indicating that silencing of E6AP may restore the activity and function of p53 protein in C33A cells.