The Mechanism Of HOTAIR/miR-126/GLS Pathway In The Malignant Progression Of Glioma

Research background and objective:Recent years,long non-coding RNA(lncRNA)has become a hot spot in the field of cancer research.As the first discovered antisense genome IncRNA,the status of HOTAIR in tumor is increasingly valued.It is generally accepted that,as a oncogene,HOTAIR can affect the transcription and expression of downstream target genes in a variety of ways.Many of these target genes are closely related to the biological behavior of tumor,such as proliferation,invasion and angiogenesis.For example,HOTAIR can inhibit the expression of miR-326,and further change the expression of fibroblast growth factor 1(FGF-1)which can affect glioma proliferation,invasion and apoptosis.But so far,the specific mechanism of HOTAIR in the development of glioma is still unknown.In this study,we analyzed the clinical data and experimental data of online database to study the effect of HOTAIR on the biological behavior of human glioma and explore its mechanism.Methods:1.The expression data of lncRNA,miRNA and mRNA in human glioma and normal brain tissue were downloaded from the Genomic Atlas of Chinese Glioma(CCGA).2.MiRNA fluorescence quantitative PCR using SYBR Green PCR reaction system and TaqMan reaction system.3.U87 and U251 cells in the logarithmic growth phase were examined by MTT assay,transwell assay and tube formation assay to detect the growth,invasion and angiogenesis of glioma cells.4.Bioinformatics prediction,Luciferase reporter assay,RT-PCR,RNA pull down and Western blot were used to verify the effect of HOTAIR on miR-126 and miR-126 on GLS targeting and to validate HOTAIR/miR-126/GLS signal pathway.Results:1.Compared with normal brain tissue,the expression of HOTAIR in glioma tissue was up-regulated.2.In vitro experiments,knockdown of HOTAIR expression can inhibit glioma cell proliferation,invasion and angiogenesis.3.Bioinformatics,Luciferase reporter assays and RNA pull down assays have shown that HOTAIR is capable of sponging endogenous miR-126.4.Compared with normal brain tissue,the expression of miR-126 in glioma tissue was down-regulated.5.In vitro studies have shown that upregulation of miR-126 can inhibit glioma cell proliferation,invasion and angiogenesis.6.Bioinformatics,luciferase reporter assay and Western Blot showed that miR-126 could bind directly to the 3’-UTR region of GLS and regulate its expression.7.Further studies have shown that elevated GLS can be partially reversed by upregulation of miR-126-induced GLS down-regulation and the resulting anti-cancer effect.8.The results show that the expression of miR-126 can be partially reversed by down-regulation of HOTAIR.Conclusion:HOTAIR/miR-126/GLS pathway plays an important role in the regulation of glioma progression.The discovery of this pathway confirms the important role of HOTAIR in the competitive endogenous RNA regulatory network,providing a new idea for the study of glioma.