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探讨多胺衍生物NNIspm诱导肝癌细胞HepG2衰老及其分子机制。采用β-半乳糖苷酶染色检测细胞衰老;应用高内涵活细胞成像系统检测细胞内NNIspm含量,细胞周期分布以及细胞内活性氧水平;Western blotting检测蛋白的表达水平;应用高效液相色谱法检测细胞内多胺含量的变化。结果表明:NNIspm可明显诱导HepG2细胞发生衰老,这与其降低细胞内多胺含量,诱导细胞周期阻滞于G0/G1期、增加细胞内活性氧有关。此外,检测到衰老标志蛋白p21的表达明显增加;相反,Cyclin E和CDK2的表达明显减弱。本研究结果表明:NNIspm引起的细胞衰老是其产生抗肿瘤作用的机制之一。
To investigate the molecular mechanism of hepatoma HepG2 induced by polyamine derivative NNIspm. Cell senescence was detected by β-galactosidase staining. Intracellular NNIspm content, cell cycle distribution and intracellular reactive oxygen species (ROS) levels were detected by high-content living cell imaging system. Western blotting was used to detect the expression of NNIspm. High performance liquid chromatography Changes in intracellular polyamine content. The results showed that NNIspm could induce HepG2 cells to senescently, which was related to the decrease of intracellular polyamines content, the induction of cell cycle arrest in G0 / G1 phase and the increase of intracellular reactive oxygen species. In addition, the expression of the aging marker protein p21 was significantly increased; on the contrary, the expressions of Cyclin E and CDK2 were significantly decreased. The results show that: NNIspm-induced cellular senescence is one of its mechanisms of anti-tumor effect.