RNA干扰PLK1基因表达对肝癌细胞增殖的影响

目的探讨靶向PLK1基因在肝癌基因治疗中的可行性。方法采用PLK1小干扰核糖核酸分子(siRNA)转染人肝癌HepG2细胞,分别以荧光实时定量PCR和Western blot检测PLK1基因和蛋白的表达水平,观察PLK1 siRNA转染对肝癌细胞体外增殖的影响。并于转染不同时间后收集细胞,分别采用琼脂糖凝胶电泳和TUNEL方法检测肝癌细胞的凋亡情况。结果 PLK1基因明显抑制癌细胞体外生长(P0.05)。肝癌HepG2细胞经siRNA转染处理后,PLK1 mRNA和蛋白表达水平明显下降(P0.05)。DNA电泳出现明显的梯度图谱;转染组癌细胞凋亡指数明显增加。结论 PLK1 siRNA转染可明显抑制肝癌细胞增殖,其机制可能与诱导细胞凋亡有关。本结果为肝癌以PLK1基因治疗提供了一条新的思路。

Polo-like kinase 1 gene silence by RNA interference inhibiting proliferation of human hepatocellular carcinoma cells

Objective:To explore the availability of targeted against polo-like kinase 1 (PLK1) gene in the therapy of hepatocellular carcinoma.
Methods:After Hep G2 hepatocellular carcinoma cells were transfected with small interfering RNA (siRNA) against PLK1, real-time RT-PCR and Western blotting were used to examine PLK1 gene expression in all cancer cells. The proliferation and growth of cancer cells in vitro were studied. Apoptosis of cancer cells was evaluated by terminal uridine deoxynucleotidyl nick end labeling (TUNEL) and agarose gel electrophoresis, respectively.
Results:Transfection of PLK1 siRNA resulted in significant inhibition of hepatocellular carcinoma cells in vitro. Expression of PLK1 in HepG2 hepatocellular carcinoma cells transfected with siRNA was down-regulated significantly. Obvious DNA ladder was shown. Cancer cells exhibited marked apoptosis in a time-dependent manner.
Conclusions:RNA interference PLK1 can inhibit proliferation through inducing apoptosis of human hepatocellular carcinoma cells. It provides a new route of treatment of hepatocellular cancer targeted against PLK1gene.