Pan Yi-Heng, Chen Jing, Sun Cui, Ma Ji-Fen, Li Xia
Center for Diagnosis and Treatment of Neuro-oncology Diseases, Taihe Hospital, Hubei University of Medicine, Shiyan 442000, Hubei, China.
Center for Diagnosis and Treatment of Neuro-oncology Diseases, Taihe Hospital, Hubei University of Medicine, Shiyan 442000, Hubei, China.
Brain Res. 2021 Mar 1;1754:147247. doi: 10.1016/j.brainres.2020.147247. Epub 2021 Jan 4.
To investigate the function of Ras-guanine nucleotide release factor 1 (Ras-GRF1) in glioma through mediating H-Ras/ERK signaling pathway.
Ras-GRF1, H-Ras, K-Ras and N-Ras expressions in glioma and normal brain tissues were detected via Immunohistochemistry. Glioma cells (U87 cells, U251 cells and primary human glioma cells) were transfected with Ras-GRF1 siRNA, H-Ras siRNA and/or Ras-GRF1 lentivirus activation particles. Then, the following aspects were evaluated: cell proliferation by MTT assay, clonogenic ability by the plate clone formation experiment, cell migration and invasion by Wound-healing and Transwell assays, and cell apoptosis by Annexin-V-FITC/PI staining. The protein expressions were measured by Western blotting. Subcutaneous and orthotopic mouse models of glioma were conducted to determine the role of Ras-GRF1 in glioma tumorigenesis.
Ras-GRF1, H-Ras, K-Ras and N-Ras expressions were upregulated in the glioma tissues, which were correlated with the WHO grade of glioma. Besides, Ras-GRF1 expression was positively related to H-Ras expression. Ras-GRF1 siRNA could reduce the expression of H-Ras and p-ERK/ERK in glioma cell. H-Ras siRNA inhibited the proliferation, clone formation, migration and invasion, and enhance the apoptosis of glioma cells, which, however, were reversed by Ras-GRF1 lentivirus activation particles. In vivo experiments also revealed that Ras-GRF1 shRNA reduced the volume and weight of the tumors in the nude mice, with down-regulations of H-Ras and p-ERK/ERK.
Ras-GRF1 was upregulated in glioma tissues and correlated with its malignancy and prognosis. Silencing Ras-GRF1, through mediating H-Ras/ERK pathway, may suppress the growth and metastasis of glioma.
通过介导H-Ras/ERK信号通路研究Ras-鸟嘌呤核苷酸释放因子1（Ras-GRF1）在胶质瘤中的作用。
采用免疫组织化学法检测胶质瘤组织和正常脑组织中Ras-GRF1、H-Ras、K-Ras和N-Ras的表达。用Ras-GRF1 siRNA、H-Ras siRNA和/或Ras-GRF1慢病毒激活颗粒转染胶质瘤细胞（U87细胞、U251细胞和原代人胶质瘤细胞）。然后，评估以下方面：通过MTT法检测细胞增殖，通过平板克隆形成实验检测克隆形成能力，通过划痕愈合和Transwell实验检测细胞迁移和侵袭，通过Annexin-V-FITC/PI染色检测细胞凋亡。通过蛋白质印迹法检测蛋白质表达。建立胶质瘤皮下和原位小鼠模型，以确定Ras-GRF1在胶质瘤肿瘤发生中的作用。
胶质瘤组织中Ras-GRF1、H-Ras、K-Ras和N-Ras表达上调，与胶质瘤的WHO分级相关。此外，Ras-GRF1表达与H-Ras表达呈正相关。Ras-GRF1 siRNA可降低胶质瘤细胞中H-Ras和p-ERK/ERK的表达。H-Ras siRNA抑制胶质瘤细胞的增殖、克隆形成、迁移和侵袭，并增强其凋亡，然而，Ras-GRF1慢病毒激活颗粒可逆转这些作用。体内实验还表明，Ras-GRF1 shRNA可降低裸鼠肿瘤的体积和重量，同时下调H-Ras和p-ERK/ERK。
Ras-GRF1在胶质瘤组织中上调，与其恶性程度和预后相关。沉默Ras-GRF1可能通过介导H-Ras/ERK通路抑制胶质瘤的生长和转移。
