Fan Fei, Yao Dongxiao, Yan Pengfei, Jiang Xiaobing, Hu Jie
Department of Neurosurgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China.
Department of Neurosurgery, General Hospital of the Yangtze River Shipping, Jiangan, Wuhan, Hubei 430010, P.R. China.
Oncol Lett. 2021 Aug;22(2):608. doi: 10.3892/ol.2021.12869. Epub 2021 Jun 15.
Glioblastoma (GBM) is the most common malignant primary brain tumor, accounting for ~57% of all gliomas and 48% of all malignant primary central nervous system tumors in the United States. Abnormal expression of the replication factor C subunit 2 (RFC2) gene and microRNA (miR)-744-5p is associated with tumorigenic characteristics, including cellular proliferation, migration and invasiveness. However, the mechanism underlying the interaction between miR-744-5p and RFC2 in GBM remains unknown. Reverse transcription-quantitative (RT-q) PCR analysis of RFC2 and miR-744-5p was performed using GBM tumor tissues and cells, and the association between miR-744-5p and RFC2 was determined by dual-luciferase reporter assay. Cell Counting Kit 8, 5-bromo-2-deoxyuridine (BrdU), wound-healing and cellular adhesion assays, as well as the detection of caspase-3 activity and western blotting were used to detect cellular proliferation, migration and adhesion, caspase-3 activity, and Bax and Bcl-2 protein expression, respectively, in GBM cells. The results of the present study demonstrated that RFC2 expression was increased in GBM tissues and cell lines. Overexpression of RFC2 promoted cellular proliferation, migration, adhesion and an increase in Bcl-2 protein levels, and suppressed cellular caspase-3 activity and Bax protein expression, while silencing RFC2 resulted in the opposite effect. The effects of miR-744-5p inhibition were similar to those of RFC2 overexpression. Moreover, miR-744-5p was found to target RFC2 in GBM cells, and inhibiting the expression of RFC2 suppressed GBM tumorigenesis. In conclusion, the present study demonstrated that miR-744-5p targets RFC2 and suppresses the progression of GBM by repressing cellular proliferation, migration and Bcl-2 protein expression, and effectively promoting caspase-3 activity and Bax protein expression. These findings suggest a new target for the clinical treatment and improved prognosis of patients with GBM in the future.
胶质母细胞瘤（GBM）是最常见的原发性恶性脑肿瘤，在美国，约占所有胶质瘤的57%以及所有原发性恶性中枢神经系统肿瘤的48%。复制因子C亚基2（RFC2）基因和微小RNA（miR）-744-5p的异常表达与肿瘤发生特征相关，包括细胞增殖、迁移和侵袭性。然而，在GBM中，miR-744-5p与RFC2之间相互作用的潜在机制仍不清楚。使用GBM肿瘤组织和细胞对RFC2和miR-744-5p进行逆转录定量（RT-q）PCR分析，并通过双荧光素酶报告基因检测法确定miR-744-5p与RFC2之间的关联。使用细胞计数试剂盒8、5-溴-2'-脱氧尿苷（BrdU）、伤口愈合和细胞黏附试验，以及检测半胱天冬酶-3活性和蛋白质印迹法，分别检测GBM细胞中的细胞增殖、迁移和黏附、半胱天冬酶-3活性以及Bax和Bcl-2蛋白表达。本研究结果表明，RFC2在GBM组织和细胞系中的表达增加。RFC2的过表达促进细胞增殖、迁移、黏附以及Bcl-2蛋白水平升高，并抑制细胞半胱天冬酶-3活性和Bax蛋白表达，而沉默RFC2则产生相反的效果。抑制miR-744-5p的作用与RFC2过表达的作用相似。此外，发现miR-744-5p在GBM细胞中靶向RFC2，抑制RFC2的表达可抑制GBM的肿瘤发生。总之，本研究表明，miR-744-5p靶向RFC2，并通过抑制细胞增殖、迁移和Bcl-2蛋白表达以及有效促进半胱天冬酶-3活性和Bax蛋白表达来抑制GBM的进展。这些发现为未来GBM患者的临床治疗和改善预后提示了一个新靶点。
