MicroRNA-154 Inhibits Migration And Invasion Of Glioma Cells By Targeting SP1

BACKGROUND&OBJECTIVE:Glioma accounts for approximately 30-40%of all central nervous system tumors and 80%of malignant brain tumors.Glioma has high mortality and high recurrence rate,despite preoperative neoadjuvant therapy and surgical resection,postoperative chemoradiotherapy and other treatment methods,the prognosis of patients with glioma is still very poor,therefore,it is urgent to develop new treatment options to improve the prognosis of patients with glioma.MicroRNAs are a class of conserved non-coding RNAs that participate in a variety of biological processes,such as cell proliferation,migration,apoptosis,and cell stem,as tumor-promoting genes or tumor suppressor genes,through complete or incomplete targets binding to the 3'-UTR region of post-transcriptional mRNA reduces translational efficiency and/or affects mRNA stability,and negative regulation plays a role in tumor suppression.SP1 plays a biological role in downstream genes by combining GC-rich sequences in target genes.More and more studies have shown that SPI regulates genes involved in tumor migration and thus affects the migration of various solid tumors.Studies have shown that SP1 is highly expressed in gliomas;we found that microRNA-154(miR-154)is under-expressed in glioma cells by reviewing the literature and combining biological prediction software,so we boldly guessed both There is a relationship between them.This study aimed to investigate whether miR-154 targets SP1 and regulates migration and invasion of glioma.METHODS:1).Human normal glial cells and 3 glioma cells were selected for real-time quantitative PCR(qRT-PCR)to detect mRNA expression levels of miR-154 and SP1,respectively.Western blot was used to detect the expression level of SP1 protein.2).Transient specific SP1 interfering RNA(si-RNA)inhibits the expression of endogenous SP1 in U87 and U251 cells,and its inhibitory potency is detected at SP1 mRNA and protein levels.Transwell experiment was used to observe SPI after interference.Changes in cell migration and invasion of U87 and U251 cells;3).Select glioma cell lines U87 and U251 as subjects,transfect miR-NC/154 mimics,and detect the expression of SP1 mRNA after overexpression of miR-154 by qRT-PCR,Western blot and transwell assays,respectively.Corresponding protein expression,cell migration and invasion ability changes;4).Using biological software and online database to predict miR-154 targeting SP1 3'-UTR region,using luciferase reporter assay to detect miR-154 target binding SP1.RESULTS:1).The expression of SP1 mRNA and protein was significantly higher in glioma cells U87,U251 and SHG44 than in normal glial cells OLN93;the expression of miR-154 was significantly higher in glioma cells U87,U251 and SHG44.Lower than normal glial cells OLN93;2).The interference of SP1 in U87 and U251 cells significantly inhibited the migration and invasion of glioma cells;3).After overexpression of miR-154 in U87 and U251,the expression of SP1 mRNA and corresponding protein decreased,and significantly inhibited cell migration and invasion ability 4).Double fluorescence experiment When the wild type vector was transfected into U87 and U251 cells,the expression was down-regulated and there was no significant change when co-transfected with the mutant vector.CONCLUSIONS:1).In glioma cells,miR-154 has low expression and SPI has high expression;2).Targeting the knockdown of SP1 to inhibit the migration and invasion of glioma cells;3)miR-154 can significantly inhibits the migration and invasion of glioma cells;4)miR-154 could inhibit the biological effects of SP1 by targeting the 3' UTR region of SP1.Therefore,high expression of SP1 plays a role in promoting the progression of glioma,and miR-154 can inhibit the progression of glioma by targeting SP1.