PDF(Pubmed)
Abstract：
Background: Berberine is one of the most interesting and promising natural anticancer drugs. POLE2 is involved in many cellular functions such as DNA replication and is highly expressed in a variety of cancers. However, the specific molecular mechanism of berberine interfering with POLE2 expression in lung adenocarcinoma (LUAD) is still unknown to a great extent. Method: The KEGG database (Release 91.0) and Gene Ontology (GO) category database were used for functional annotation of differentially expressed genes after berberine treatment. Reproducibility assessment using TCGA dataset. The biological functions of berberine in LUAD were investigated by a series of in vitro and in vivo experiments: MTT, colony formation, mouse xenograft and plasmid transfection. The molecular mechanisms of berberine were demonstrated by plasmid transfection, quantitative RT-PCR and Western blotting. Result: The elevated expression of FOXM1 and the high enrichment of DNA replication pathway were confirmed in LUAD by microarray and TCGA analysis, and were positively correlated with poor prognosis. Functionally, berberine inhibited the proliferation and survival of LUAD cell lines in vitro and in vivo. Mechanistically, berberine treatment down regulated the expression of FOXM1which closely related to survival, survival related genes in Cell cycle and DNA replication pathway, and significantly down regulated the expression of survival related POLE2. Interestingly, we found that the transcription factor FOXM1 could act as a bridge between berberine and POLE2. Conclusion: Berberine significantly inhibited LUAD progression via the FOXM1/POLE2, and FOXM1/POLE2 may act as a clinical prognostic factor and a therapeutic target for LUAD. Berberine may be used as a promising therapeutic candidate for LUAD patients.
摘要：
背景:小檗碱是最有前景的天然抗癌药物之一。POLE2参与许多细胞功能，如DNA复制，并在多种癌症中高度表达。然而,小檗碱干扰POLE2在肺腺癌(LUAD)中表达的具体分子机制尚不清楚。方法:采用KEGG数据库(Release91.0)和基因本体(GeneOntology,GO)分类数据库对黄连素处理后的差异表达基因进行功能注释。使用TCGA数据集进行重复性评估。通过一系列体外和体内实验研究了小檗碱在LUAD中的生物学功能：MTT，菌落形成,小鼠异种移植和质粒转染。通过质粒转染证实小檗碱的分子机制,定量RT-PCR和Western印迹。结果：基因芯片和TCGA分析证实了LUAD中FOXM1的表达升高和DNA复制途径的高度富集。与不良预后呈正相关。功能上,小檗碱在体外和体内抑制LUAD细胞系的增殖和存活。机械上,黄连素治疗下调与生存密切相关的FOXM1的表达,细胞周期和DNA复制途径中的生存相关基因，并显著下调生存相关POLE2的表达。有趣的是,我们发现转录因子FOXM1可以作为小檗碱和POLE2之间的桥梁。结论:小檗碱通过FOXM1/POLE2显著抑制LUAD的进展,FOXM1/POLE2可作为LUAD的临床预后因素和治疗靶点。小檗碱可用作LUAD患者的有希望的治疗候选物。
