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目的:探讨质粒表达载体介导的RNA干扰抑制PLK1基因表达对人食管鳞癌细胞Eca-109的P53基因的影响。方法:构建靶向人PLK1基因的RNA干扰表达质粒(PLK1-shRNA),将该干扰质粒转染到人食管鳞癌细胞Eca-109中,采用Westernblot技术分析检测RNA干扰后48h、72h各组细胞PLK1基因和P53基因蛋白表达水平的变化。结果:Westernblot结果显示,转染RNA干扰表达质粒(PLK1-shRNA)后,PLK1-shRNA组Eca-109细胞48h、72h较转染液对照组的PLK1基因蛋白表达分别降低了83.7%和56.0%(P<0.05);PLK1-shRNA组Eca-109细胞48h、72h较转染液对照组的P53基因蛋白表达分别升高了46.0%和70.5%(P<0.05)。结论:RNA干扰PLK1基因能够有效地抑制Eca-109细胞中PLK1基因的表达,从而有效地升高P53基因的表达,说明PLK1基因抑制P53基因在Eca-109细胞中的表达。
AIM: To investigate the effect of plasmid-mediated RNA interference (RNAi) on P53 gene expression in human esophageal squamous carcinoma cell line Eca-109 induced by PLK1 gene. Methods: The PLK1-shRNA targeting human PLK1 gene was constructed and transfected into human esophageal squamous carcinoma cell line Eca-109. Western blotting was used to detect the expression of PLK1-shRNA at 48 and 72 h PLK1 gene and P53 gene protein expression level changes. Results: The results of Western blot showed that PLK1-shRNA expression in PLK1-shRNA group decreased by 83.7% and 56.0% (P <0.05) compared with the control group at 48h and 72h after transfected with PLK1-shRNA P <0.05). The expression of P53 protein in Eca-109 cells increased by 46.0% and 70.5% (P <0.05) at 48h and 72h compared with the control group. Conclusion: RNA interference of PLK1 gene can effectively suppress the expression of PLK1 gene in Eca-109 cells and effectively increase the expression of P53 gene, indicating that PLK1 gene inhibits the expression of P53 gene in Eca-109 cells.