Departments of Biology, Faculty of Science, Malayer University, Malayer, Iran.
Integrative Oncology Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR, Tehran, Iran.
Life Sci. 2021 Feb 1;266:118874. doi: 10.1016/j.lfs.2020.118874. Epub 2020 Dec 22.
Hypericin (HYP) from Hypericum perforatum has cytotoxic effects on a variety of malignant cell types, but the pattern of gene expression mediating the effect is largely unknown. Here we sought to analyze the response of U87 glioblastoma (GBM) cell lines in response to HYP.
U87 cell line was treated by HYP. Cytotoxicity was assessed using MTT and Annexin V/PI assays. Gene expression profile was obtained using high-throughput sequencing. Enrichment analysis was performed on differentially expressed genes (DEGs). Upstream transcription factors and microRNAs regulating DEGs were predicted. The effects of DEGs on survival of GBM patients were calculated. Protein-protein interaction analysis was conducted to obtain key altered genes. The possible effect of HYP treatment on immunity response was evaluated.
The IC50 of HYP on U87 cell line was determined to be 1.5 μg/ml. The main type of cell death was apoptosis. A total of 312 DEGs were found. Affected Gene Ontology terms and pathways were identified. Analysis of upstream modulators of DEGs pointed out to transcription factors that significantly overlap with GBM stem cell transcription factor. Survival analysis suggested that HYP works best for the mesenchymal subtype patients. Tumor infiltration analysis predicted that HYP may affect Treg and macrophage infiltration in vivo. Using expression pattern of GBM patients and HYP-induced DEGs we suggested Fedratinib as a complementary drug to HYP.
Our study represents the response of U87 cell line to HYP, with analyses on survival, transcription factors and personalization according to GBM subtype.
贯叶金丝桃（HYP）具有细胞毒性作用，可杀伤多种恶性细胞类型，但介导这种作用的基因表达模式在很大程度上尚不清楚。本研究旨在分析 U87 神经胶质瘤（GBM）细胞系对 HYP 的反应。
用 HYP 处理 U87 细胞系。采用 MTT 和 Annexin V/PI 法评估细胞毒性。采用高通量测序获得基因表达谱。对差异表达基因（DEGs）进行富集分析。预测调控 DEGs 的上游转录因子和 microRNAs。计算 DEGs 对 GBM 患者生存的影响。进行蛋白质-蛋白质相互作用分析以获得关键改变基因。评估 HYP 治疗对免疫反应的可能影响。
确定 HYP 对 U87 细胞系的 IC50 为 1.5μg/ml。主要的细胞死亡类型为细胞凋亡。共发现 312 个 DEGs。确定了受影响的 GO 术语和途径。分析 DEGs 的上游调节剂指出，转录因子与 GBM 干细胞转录因子显著重叠。生存分析表明，HYP 对间充质亚型患者效果最佳。肿瘤浸润分析预测 HYP 可能影响体内 Treg 和巨噬细胞浸润。利用 GBM 患者的表达模式和 HYP 诱导的 DEGs，我们建议 Fedratinib 作为 HYP 的补充药物。
本研究代表了 U87 细胞系对 HYP 的反应，根据 GBM 亚型进行了生存、转录因子和个体化分析。
