Organotypic 3D cell-architecture architecture impacts the expression pattern of miRNAs-network in breast cancer SKBR3 cells
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PROVIDER: PRJNA1001008 | ENA |
REPOSITORIES: ENA
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Project description:Organotypic 3D cell-architecture architecture impacts the expression pattern of miRNAs-network in breast cancer SKBR3 cells [miRNA-seq]
| PRJNA1001896 | ENA
Project description:BackgroundCurrently, most of the research on breast cancer has been carried out in conventional two-dimensional (2D) cell cultures due to its practical benefits, however, the three-dimensional (3D) cell culture is becoming the model of choice in cancer research because it allows cell-cell and cell-extracellular matrix (ECM) interactions, mimicking the native microenvironment of tumors in vivo.MethodsIn this work, we evaluated the effect of 3D cell organization on the expression pattern of miRNAs (by Small-RNAseq) and mRNAs (by microarrays) in the breast cancer SKBR3 cell line and analyzed the biological processes and signaling pathways regulated by the differentially expressed protein-coding genes (DE-mRNAs) and miRNAs (DE-microRNAs) found in the organoids.ResultsWe obtained well-defined cell-aggregated organoids with a grape cluster-like morphology with a size up to 9.2 × 105 μm3. The transcriptomic assays showed that cell growth in organoids significantly affected (all p < 0.01) the gene expression patterns of both miRNAs, and mRNAs, finding 20 upregulated and 19 downregulated DE-microRNAs, as well as 49 upregulated and 123 downregulated DE-mRNAs. In silico analysis showed that a subset of 11 upregulated DE-microRNAs target 70 downregulated DE-mRNAs. These genes are involved in 150 gene ontology (GO) biological processes such as regulation of cell morphogenesis, regulation of cell shape, regulation of canonical Wnt signaling pathway, morphogenesis of epithelium, regulation of cytoskeleton organization, as well as in the MAPK and AGE-RAGE signaling KEGG-pathways. Interestingly, hsa-mir-122-5p (Fold Change (FC) = 15.4), hsa-mir-369-3p (FC = 11.4), and hsa-mir-10b-5p (FC = 20.1) regulated up to 81% of the 70 downregulated DE-mRNAs.ConclusionThe organotypic 3D cell-organization architecture of breast cancer SKBR3 cells impacts the expression pattern of the miRNAs-mRNAs network mainly through overexpression of hsa-mir-122-5p, hsa-mir-369-3p, and hsa-mir-10b-5p. All these findings suggest that the interaction between cell-cell and cell-ECM as well as the change in the culture architecture impacts gene expression, and, therefore, support the pertinence of migrating breast cancer research from conventional cultures to 3D models.
| S-EPMC10661268 | biostudies-literature
Project description:The transcriptomic assays showed that cell growth in organoids significantly affected (all p < 0.01) the gene expression patterns of both, miRNAs and mRNAs, finding 20 upregulated and 19 downregulated DE-microRNAs, as well as 49 upregulated and 123 downregulated DE-mRNAs. In silico analysis showed that a subset of 11 upregulated DE-microRNAs target 70 downregulated DE-mRNAs. These genes are involved in 150 gene ontology (GO) biological processes such as regulation of cell morphogenesis, regulation of cell shape, regulation of canonical Wnt signaling pathway, morphogenesis of epithelium, regulation of cytoskeleton organization, as well as in the MAPK and AGE-RAGE signaling KEGG-pathways. Interestingly, hsa-mir-122-5p (Fold Change (FC)=15.4), hsa-mir-369-3p (FC=11.4), and hsa-mir-10b-5p (FC=20.1) regulated up to the 81 % of the 70 downregulated DE-mRNAs
2023-08-01 | GSE239813 | GEO
Project description:The transcriptomic assays showed that cell growth in organoids significantly affected (all p < 0.01) the gene expression patterns of both, miRNAs and mRNAs, finding 20 upregulated and 19 downregulated DE-microRNAs, as well as 49 upregulated and 123 downregulated DE-mRNAs. In silico analysis showed that a subset of 11 upregulated DE-microRNAs target 70 downregulated DE-mRNAs. These genes are involved in 150 gene ontology (GO) biological processes such as regulation of cell morphogenesis, regulation of cell shape, regulation of canonical Wnt signaling pathway, morphogenesis of epithelium, regulation of cytoskeleton organization, as well as in the MAPK and AGE-RAGE signaling KEGG-pathways. Interestingly, hsa-mir-122-5p (Fold Change (FC)=15.4), hsa-mir-369-3p (FC=11.4), and hsa-mir-10b-5p (FC=20.1) regulated up to the 81 % of the 70 downregulated DE-mRNAs
2023-08-04 | GSE239998 | GEO
Project description:Long noncoding RNA plays a significant role in gene regulation. Long noncoding RNA TMS1AS negatively regulates the sense strand gene TMS1 in breast cancer cells SKBR3. TMS1 is invovled in cell growth and apoptosis through regulating its target genes. We used microarrays to detail the global programme of gene expression upon TMS1AS knockdown or TMS1 knockdown and analyzed the specifity of TMS1AS regulating TMS1.
2017-08-01 | GSE85032 | GEO
Project description:SKBR3 cells expressing NDRG1 shRNA1 or vector control were harvested by trypsinization and total RNA was extracted. Silencing NDRG1 reduces cell proliferation rates, causing lipid metabolism dysfunction including increased fatty acid incorporation into neutral lipids and lipid droplets. global changes in transcriptome due to NDRG1 silencing were observed
2018-06-21 | GSE112841 | GEO
Project description:Organs consist of not only parenchyma but also stroma, the latter of which coordinates generation of organotypic structures. Despite recent advances in organoid technology, induction of organ-specific stroma and recapitulating the complex organ configurations from pluripotent stem cells (PSCs) has remained challenging. For the kidney, the stromal progenitor coordinates differentiation of the two parenchymal progenitors: nephron progenitor and ureteric bud. By identifying the origin and dorsoventral patterning of the renal stromal progenitor, we established its induction protocol from mouse PSCs. When the three types of progenitors were differentially induced from PSCs and assembled, the all PSC-derived organoids reproduced the complex kidney structures, with multiple types of stromal cells distributed around differentiating nephrons and branching ureteric buds. Thus, integration of PSC-derived stroma into the conventional organoids enables recapitulation of organotypic architecture.
2021-12-05 | GSE178263 | GEO
Project description:Low NTN4 expression is correlated with breast cancer proliferation and metastasis, while the molecular mechanism of NTN4 has not been fully explored. Here, the breast cancer cells SKBR3 stably expressing dCas9-VP64 and MS-P65-HSG1 were generated by transducing a lentiviral plasmid containing lenti-dCAS-VP64-Blast (Addgene plasmid # 61425) and lenti-EF1a-MS2–p65–HSF1-2A-Hygro-WPRE (Addgene plasmid # 89308), followed by selection and maintained using 10 μg/mL of blasticidin and 200 μg/mL of hygromycin. SKBR3-dCas9-VP64-MPH cells were further transduced with a doxycycline (Dox)-induced expressing sgRNA targeting NTN4 promoter. The successfully transfected cells were selected and maintained with puromycin at a concentration of 0.5 μg/mL for SKBR3-dCas9-VP64-MPH. SKBR3-dCas9-VP64-MPH-AC-sgRNA cells were treated with or without 50 ng/μL Dox (three biological repeats for both groups) to induce NTN4 overexpressing. Total RNA was extracted using TRIzol according to the manufacturer’s instructions. The quality of RNA samples was examined by Agilent BioAnalyser 2100. The samples were subjected to RNA-sequencing analysis on the BGISEQ-500 system by Beijing Genomics Institute (BGI, China). The clean-tag reads were aligned to the reference genome and reference genes using HISAT2 and Bowtie2. The matched reads were calculated and then normalized using RSEM software. Differential gene expression was analyzed using DESeq2 and was carried out further to explore the mechanism of NTN4 in breast cancer.
2022-05-27 | GSE197635 | GEO
Project description:To explore TSA influence on human breast cancer cells, we attempt to analyze genes differentially expressed between TSA treated and untreated SKBR3 cells, which will hopefully provide clues for TSA target genes.
2017-02-28 | GSE95087 | GEO
Project description:Proteomic of SKBr3 cells of Homo sapiens treated by PFOA
2020-05-26 | PXD019397 |