Project description:Tremendous studies have found that the abnormality of long noncoding RNA (lncRNA) contributed to cancer initiation, progression, and recurrence via multiple signaling cascades. Nevertheless, the possible underlying mechanisms of lncRNA in temozolomide (TMZ)-resistant glioma were not well understood, hindering the improvement of TMZ-based therapies against glioma. The present study illustrated that the lncRNA KCNQ1OT1 increased in TMZ-resistant gliomas cells compared to the parental cells. Introduction of KCNQ1OT1 boosted glioma cell viability, clonogenicity and rhodamine 123 efflux while hampered apoptosis post-exposure to TMZ. Consistent with bioinformatic prediction, KCNQ1OT1 directly sponged miR-761, which was downregulated in TMZ-resistant glioma cell lines. Overexpression of miR-761 attenuated glioma cell viability and clonogenicity while triggered apoptosis and rhodamine 123 cellular accumulation post-exposure to TMZ, leading to rehabilitated glioma TMZ-sensitivity, which was against the function of KCNQ1OT1. miR-761 bound to 3’-untranslated region of PIM1, a proto-oncogene with constitutive serine/threonine kinase activity, attenuated PIM1-mediated signaling cascades. Furthermore, stable knockdown of KCNQ1OT1 by small hairpin RNA amplified the TMZ-induced tumor regression in TMZ-resistant U251 mouse models. Briefly, the present study evaluated KCNQ1OT1 conferred TMZ resistance by sponging miR-761 and releasing PIM1 expression, resulting in activation of PIM-mediated MDR1/c-Myc/Survivin signaling pathways. The findings mentioned above extended the knowledge of lnRNA KCNQ1OT1 in the regulation of chemoresistance in glioma and provided a promising therapeutic target for TMZ-resistant glioma patients. Long noncoding RNA profiling by array
Project description:To identify TGF-β regulated lncRNAs in glioblastoma, we performed a genome-wide microarray screen in T98G glioma cells. T98G cells were treated with 10 ng/ml TGF-β (24h) and differentially expressed lncRNAs were identified using microarray in comparison with control cells.
Project description:Glioblastomas show heterogeneous histological features. These distinct phenotypic states are thought to be associated with the presence of glioma stem cells (GSCs), which are highly tumorigenic and self-renewing sub-population of tumor cells that have different functional characteristics. We found that TUG1, a long non-coding RNA (lncRNA), plays pivotal roles in maintaining the self-renewal properties of GSC. Some lncRNAs are known to act as a miRNA sponge in the cytoplasm, where lncRNAs bind to miRNAs and quench their activity. To investigate miRNA expression profiling in GSC upon TUG1 inhibition, we have performed microarray experiment using SurePrint G3 Human miRNA 8x60K Microarray (G4872A, Agilent Technologies).
Project description:FOXM1 plays a key role in M phase in normal cells and is overexpressed in human glioma. We found that FOXM1 deprivation could sensitize the glioma cells to TMZ chemotherapy. To find out the mechanistic regulation of FOXM1 in chemo-resistant genes, we used microarrays to select the potential genes regulated by FOXM1 which dominates in glioma chemo-resistance. U87 glioma cells were transiently transfected with none-target siRNA or FOXM1 siRNA. Total RNA were extracted after 48 hours and subjected to the microarray.
Project description:To investigate the potential pathogenic mechanism of glioma-related epilepsy (GRE), we have employed analyzing of the dynamic expression profiles of microRNA/ mRNA/ lncRNA in brain tissues of glioma patients. Brain tissues of 16 patients with GRE and nine patients with glioma without epilepsy (GNE) were collected. The total RNA was dephosphorylated, labeled, and hybridized to the Agilent Human miRNA Microarray, Release 19.0, 8x60K. The cDNA was labeled and hybridized to the Agilent LncRNA+mRNA Human Gene Expression Microarray V3.0, 4x180K. The raw data was extracted from hybridized images using Agilent Feature Extraction, and quantile normalization was performed using the Agilent GeneSpring. We found that three differentially expressed miRNAs (miR-10a-5p, miR-10b-5p, miR-629-3p), six differentially expressed lncRNAs (TTN-AS1, LINC00641, SNHG14, LINC00894, SNHG1, OIP5-AS1), and 49 differentially expressed mRNAs may play a vitally critical role in developing GRE.
Project description:Dysregulated long noncoding RNAs (lncRNAs) are involved in the pathogenesis and development of human diseases, such as epithelial ovarian cancer (EOC). In this study, we identified EOC-related lncRNAs and performed lncRNA and mRNA microarray analyses using IOSE80, a normal ovary cell line, and two ovarian carcinoma cell lines (SKOV3 and SKOV3/DDP) to investigate the potential roles of lncRNAs in EOC. lncRNA-HEIH expression in EOC tissues and cell lines was measured by quantitative real-time polymerase chain reaction (qPCR). In addition, we generated a lncRNA-mRNA co-expression network in order to identify lncRNA-expression trends and potential lncRNA target genes. Cell viability, migration, and invasion were determined by Cell Counting Kit-8, transwell assay, and wound-healing assay, respectively, and apoptosis was analyzed by flow cytometry. We identified 3527 differentially expressed lncRNAs upon comparison of the lncRNA profiles from IOSE80 with those of SKOV3 cell lines, with 11 differentially expressed lncRNAs confirmed by qPCR. Both pathway and gene ontology analyses demonstrated the involvement of lncRNAs, especially HEIH and LINC-PINT, in multiple biological processes. Furthermore, in vitro knockdown experiments confirmed that suppression of HEIH expression inhibited EOC cell proliferation. Our findings provide a foundation for further research into the role of these lncRNAs in EOC carcinogenesis and progression.