E2F1转录激活SKA1在促进肝癌细胞增殖中的作用及机制研究

Study on the effect and mechanism of SKA1 activated by E2F1 transcription in promoting the proliferation of liver cancer cells

目的本文旨在研究SKA1是否是调节肝癌恶性增殖的一个关键分子,并进一步探究其机制,为后续靶向治疗提供分子理论基础。方法通过生物信息学技术分析来自TCGA数据库中的肝癌数据,分析了SKA1在肝癌中的表达量,同时,我们还分析了SKA1的表达与肝癌患者预后的关系。采用基于脂质体介导的细胞转染技术,构建过表达SKA1的肝癌细胞系,并进一步采用CCK-8及平板克隆试验检测SKA1对肝癌细胞增殖能力的影响。采用生物信息学技术对SKA1及E2F1表达相关性进行分析,进一步检测E2F1在SKA1启动子区域的结合位点,并采用双荧光素酶技术检测E2F1对SKA1的转录激活作用。结果SKA1在肝癌组织中高表达,且SKA1高表达的肝癌患者的总体生存率(49.8%)比SKA1低表达患者低(69.4%),二者呈负相关。与此同时,E2F1也在肝癌组织中呈高表达,且E2F1高表达肝癌患者生存期显著低于低表达组,与不良预后呈负相关。SKA1过表达可将肝癌细胞的增殖能力提升近50%,SKA1受E2F1转录因子调控,且E2F1转录因子与SKA1启动子结合,转录激活SKA1在肝癌细胞中的表达。结论E2F1转录激活SKA1促进肝癌细胞增殖,导致肝癌患者预后不良。

Objective To investigate whether SKA1 is a key molecule regulating malignant proliferation of liver cancer,and further explore its mechanism to provide molecular theoretical basis for subsequent targeted therapy.Methods The data of liver cancer from TCGA database were analyzed by bioinformatics technology.The expression of SKA1 in liver cancer was analyzed.At the same time,we also analyzed the relationship between the expression of SKA1 and the prognosis of patients with liver cancer.The hepatoma cell line overexpressing SKA1 was constructed by liposome-mediated cell transfection technique,and the effect of SKA1 on the proliferation of hepatoma cells was further tested by CCK-8 and plate cloning assay.At the same time,we found that E2F1 is also highly expressed in liver cancer,using bioinformatics technology to analyze the correlation between SKA1 and E2F1 expression,further detecting the binding site of E2F1 in the SKA1 promoter region,and using dual luciferase technology to detect E2F1 against SKA1.Transcriptional activation.Results KA1 was highly expressed in liver cancer tissues,and the overall survival rate of liver cancer patients with high SKA1 expression was 49.8%,lower than that of patients with low SKA1 expression,showing a negative correlation.E2F1 is also highly expressed in liver cancer tissues,and the survival time of patients with liver cancer with high E2F1 expression is significantly lower than that in the low expression group,which was negatively correlated with poor prognosis.SKA1 overexpression could increase the proliferation ability of liver cancer cells by nearly 50%.SKA1 is regulated by the E2F1 transcription factor,and the E2F1 transcription factor is combined with the SKA1 promoter to transcriptionally activate the expression of SKA1 in liver cancer cells.Conclusion E2F1 transcriptional activation of SKA1 promotes proliferation of hepatoma cells,leading to poor prognosis in patients with liver cancer.