Project description:HOTAIR was found to be overepressed in a subset of urothelial cancer tissues and cell lines compared to normal controls. Ectopic HOTAIR expression in urothelial cancer cells in vitro demonstrated cell type dependent changes in phenotype. While some cell lines showed increased proloferation activity and induction of an aggressive phenotypes (e.g. stable transfected VM-CUB1 cells), others displayed rather a reduction of proliferation and migration. Stable transfection of 5637 cells resulted in induction of an immune response. Results of microarray analysis of stable transfected VM-CUB1 and 5637 cells concurred well with observed phenotypical cell type-specific changes. For differential gene expression analyses three independent high quality RNA preparations from VM-CUB1 cells, stably transfected with HOTAIR (clone 20), and 5637 cells, stably transfected with HOTAIR (clone 4), were compared to the respective vector control cells.
Project description:The human HOTAIR-specific siRNA and control siRNA were transfected into MCF-7-TNR cells. RNA was collected at 72 hrs after transfection. RNA-SEQ was carried out to profile the gene expression altered by HOTAIR knockdown.
Project description:To analysis of the effect of HOTAIR on global DNA methylation in KYSE180 esophageal cancer cell line, genome-scale DNA methylation was analyzed in a pair of esophageal cancer cell lines, KYSE180 cells stably transfected with HOTAIR and control KYSE180 cells. The results from genome-wide DNA methylation analysis indicate that hypermethylation occurs more frequently than hypomethylation after stably ectopic expressing HOTAIR in KYSE180 cells.
Project description:To investigate the functional role of HOTAIR in esophageal squanmous cell carcinoma (ESCC) cells, RNAi-mediated knockdown of HOTAIR and overexpression of HOTAIR were carried out using KYSE180 cells. Gene expression microarray analysis revealed that a number of reported HOTAIR target genes were regulated by HOTAIR. Moreover, gene ontology analysis revealed enrichment of genes closely related to tumorigenesis, such as cell migration, regulation of cell cycles. For RNAi-mediated knockdown of HOTAIR, two different stealth siRNAs against HOTAIR were generated by Invitrogen, after which a mixture of the two was used for transfection. KYSE180 cells in 6-well plates were transfected with 100 pmol of Stealth siRNA (Invitrogen) or a Stealth RNAi Negative Control Medium GC (Invitrogen) using Lipofectamine2000 (Invitrogen). For HOTAIR overexpression, KYSE180 cells were transfected with LZRS_HOTAIR vector using Polyfect Transfection Reagent (Qiagen). Total RNA was extracted 48 h after transfection.
Project description:HOTAIR was found to be overepressed in a subset of urothelial cancer tissues and cell lines compared to normal controls. Ectopic HOTAIR expression in urothelial cancer cells in vitro demonstrated cell type dependent changes in phenotype. While some cell lines showed increased proloferation activity and induction of an aggressive phenotypes (e.g. stable transfected VM-CUB1 cells), others displayed rather a reduction of proliferation and migration. Stable transfection of 5637 cells resulted in induction of an immune response. Results of microarray analysis of stable transfected VM-CUB1 and 5637 cells concurred well with observed phenotypical cell type-specific changes.
Project description:LincRNA HOTAIR was expressed in pure SCLC and higher expression was significantly related to lymphatic invasion and relapse. Multivariate analyses demonstrated that HOTAIR expression correlated with RFS. In vitro experiments demonstrated that half of SCLC cell lines expressed HOTAIR at higher levels than normal cells and that, using cells of SBC-3, knockdown of HOTAIR decreased proliferative activity and cellular invasiveness with altered expression of cell adhesion-related genes. Total RNAs from one siHOTAIR-transfected SBC-3 cells and control cells (siGFP-transfected cells), were extracted using RNeasy mini kit Plus (Qiagen) and hybridized to the microarrays, Sure Print G3 Human GE 8x60K microarrays (Agilent Technologies, Santa Clara, CA, USA), according to the manufacturerM-bM-^@M-^Ys instructions. Subsequently, data analysis was carried out using the GeneSpring GX 12 software (Agilent Technologies), with a stringency of P<0.1 and a 2-fold or more change using gene ontology analysis.
Project description:To analyze the effect of HOTAIR depletion in a gastrointestinal stromal tumor (GIST) cells, RNAi-mediated knockdown of HOTAIR was carried out using GIST-T1 cells. Gene expression microarray analysis revealed that a number of reported HOTAIR target genes were upregulated by knockdown of HOTAIR. Moreover, we found that 1424 genes were upregulated by siHOT (>2-fold), and Gene Ontology analysis revealed enrichment of genes related to “nucleus”, “chromosome” and “membrane-bounded organelle.” For RNAi-mediated knockdown of HOTAIR, three different Stealth siRNAs against HOTAIR were generated by Invitrogen, after which a mixture of the three was used for transfection. GIST-T1 cells in 6-well plates were transfected with 100 pmol of Stealth siRNA (Invitrogen) or a Stealth RNAi Negative Control Medium GC (Invitrogen) using Lipofectamine2000 (Invitrogen). Total RNA was extracted 48 h after transfection.
Project description:Renal cell carcinoma (RCC) is one of the most lethal urologic cancers. About one-third of RCC patients already have distal metastasis at the time of diagnosis. There is growing evidence that Hox antisense intergenic RNA (HOTAIR) plays essential roles in metastasis in several types of cancers. However, the precise mechanism by which HOTAIR enhances malignancy remains unclear, especially in RCC. Here, we demonstrated that HOTAIR enhances RCC-cell migration by regulating the insulin growth factor-binding protein 2 (IGFBP2) expression. HOTAIR expression in tumors was significantly correlated with nuclear grade, lymph-node metastasis, and lung metastasis. High HOTAIR expression was associated with a poor prognosis in both our dataset and The Cancer Genome Atlas dataset. Migratory capacity was enhanced in RCC cell lines in a HOTAIR-dependent manner. HOTAIR overexpression accelerated tumorigenicity and lung metastasis in immunodeficient mice. Microarray analysis revealed that IGFBP2 expression was upregulated in HOTAIR-overexpressing cells compared with control cells. The enhanced migration activity of HOTAIR-overexpressing cells was attenuated by IGFBP2 knockdown. IGFBP2 and HOTAIR were co-expressed in clinical RCC samples. Our findings suggest that the HOTAIR-IGFBP2 axis plays critical roles in RCC metastasis and may serve as a novel therapeutic target for advanced RCC.