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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(P<0.05)。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(P<0.05)。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。
Objective To investigate the feasibility of targeting PLK1 gene in hepatocellular carcinoma gene therapy. Methods HepG2 cells were transfected with PLK1 small interfering RNA (siRNA). The expression of PLK1 gene and protein was detected by real-time PCR and Western blot respectively. The effect of PLK1 siRNA on the proliferation of HepG2 cells was observed. The cells were harvested at different times of transfection. The apoptosis of hepatoma cells was detected by agarose gel electrophoresis and TUNEL method respectively. Results PLK1 gene significantly inhibited the growth of cancer cells in vitro (P <0.05). After transfection with HepG2 cells, the expression of PLK1 mRNA and protein were significantly decreased (P <0.05). DNA electrophoresis showed a clear gradient map; transfected cells significantly increased apoptotic index. Conclusion PLK1 siRNA transfection can significantly inhibit the proliferation of hepatocellular carcinoma cells, which may be related to the induction of apoptosis. This result provides a new idea for PLK1 gene therapy for liver cancer.