论文部分内容阅读
背景与目的:Aes(amino-terminal enhancer of split,Aes)是一种能够与转录因子结合调节转录活性的蛋白,以往的研究显示其在发育过程中起重要作用,近期研究发现Aes可参与结肠癌转移。本研究旨在观察外源Aes在肝癌细胞中的定位以及其对肝癌细胞增殖、侵袭和迁移能力的影响。方法:构建pEGFP-Aes表达载体,并通过LipofectamineTM2000将其转入Aes低表达的肝癌细胞系HepG2中,蛋白质印迹法(Westernblot)检测外源Aes的表达,荧光显微镜观察pEGFP-Aes在细胞中的定位,应用细胞计数盒(Cell CountingKit)-8检测转染Aes后细胞增殖活力变化;划痕实验检测转染前后细胞迁移能力的变化;Transwell实验检测Aes对细胞侵袭能力的影响。结果:Aes在HepG2中表达较低,将Aes转入肝癌细胞系HepG2,Aes在细胞核和细胞质中都有表达,并且在细胞核中形成点状浓积,Aes对肝癌细胞HepG2的增殖无显著影响,但能抑制HepG2的侵袭和迁移能力(P<0.05)。结论:Aes对肝癌细胞恶性表型的影响主要是抑制其侵袭和迁移能力。
BACKGROUND AND AIM: Aes (Aes), a protein that binds to transcription factors and regulates transcriptional activity, has been shown to play an important role in development. A recent study found that Aes participates in colon cancer Transfer. This study aimed to observe the localization of exogenous Aes in hepatoma cells and its effect on the proliferation, invasion and migration of hepatocellular carcinoma cells. Methods: The pEGFP-Aes expression vector was constructed and transfected into HepG2 cell line with low expression of Aes by LipofectamineTM2000. The expression of exogenous Aes was detected by Western blotting. The localization of pEGFP-Aes in cells was observed by fluorescence microscopy . Cell Counting Kit-8 was used to detect the changes of cell proliferation after transfected with Aes. Scratch assay was used to detect the changes of cell migration ability before and after transfection. Transwell assay was used to detect the effect of Aes on cell invasiveness. Results: The expression of Aes in HepG2 was low. Aes was transfered into HepG2 cell line. The expression of Aes in nucleus and cytoplasm, and the formation of punctate accumulation in nucleus, Aes had no significant effect on the proliferation of HepG2 cells. But inhibited the invasion and migration ability of HepG2 (P <0.05). Conclusion: The effect of Aes on the malignant phenotype of hepatocellular carcinoma cells is mainly to inhibit its invasion and migration.