Project description:To gain insight into the differential gene expression to aberrant fibroblast activation of the TME, we isolated primary CAFs and NFs from human NSCLC tissues and paired normal tissues,and performed microarray screening of CAFs and NFs
Project description:To identify miRNA differentially expressed in CAFs vs matched NFs and analyze the heterogeneity of miRNA expression profiles in the two kinds of cells, we established primary cultures of CAFs and paired NFs from six resected breast tumor tissue without any radiotherapy and chemotherapy treatment.
Project description:Transcriptional profiling of human carcinoma-associated fibroblasts (CAFs) comparing control normal fibroblasts (NFs). NFs derived from normal tissues and CAFs derived from the patients with oral cancer were identified by immunocytochemistry. Goal was to determine differentially expressed lncRNAs between NFs and CAFs.
Project description:To understand the driving mechanisms for CAF activation, we performed H3K27ac and H3K4me1 Cleavage Under Targets and Tagmentation (CUT&Tag) analysis in six pairs of NFs and CAFs. At H3K4me1+ sites, we identified 3,333 regions with H3K27ac levels in CAFs ≥ 2-fold than matched NFs in more than 3 pairs, and defined them as CAF-activated enhancers.
Project description:Although how CAFs impact the tumor epithelium is being progressively unveiled, transcriptional processes of lineage plasticity in fibroblasts govern the acquisition and transition of the CAF phenotype are much less understood. Here, to explore the potential involvement of the circular RNAs (circRNAs) in fibroblast activation and phenotype acquisition, we isolated CAFs and NFs from human pancreatic cancer and paired normal pancreatic tissue. Next, we conducted circRNA profiling five CAFs and three paired NFs by high throughput sequencing. Our results showed that clusters of circRNAs were aberrantly expressed in CAFs compared with NFs, and provided potential targets for future treatment of PDAC and novel insights into PDAC microenvironment.
Project description:To identify miRNA differentially expressed in CAFs vs matched NFs and analyze the heterogeneity of miRNA expression profiles in the two kinds of cells, we established primary cultures of CAFs and paired NFs from six resected breast tumor tissue without any radiotherapy and chemotherapy treatment. Paired CAFs and NFs from six primary human breast carcinoma specimens were cultured,the third passage of primary cells was used in the experiments.
Project description:This study aims to identify genes differentially expressed in oral CAFs with respect to their normal counterparts, i.e. oral NFs. We collected oral normal mucosal tissues and oral squamous cell carcinoma (OSCC) tissues from 12 healthy people and 12 OSCC patients, respectively, at West China Hospital of Stomatology, Sichuang University, P. R. China. The tissues were isolated, cultured and purified using standard protocols to obtain oral NFs and CAFs. Agilent 4×44K Whole Genome Oligo Microarrays were used to measure the gene expression profiles of the samples. Sample preparation and hybrid reactions were performed according to the manufacture’s instructions. RNA samples from NFs and CAFs were pooled respectively before hybridization. The chips were scanned and visualized using the Agilent DNA microarray scanner. Probe set intensities were measured using Agilent Feature Extraction software (version 10.5.1.1). Raw data were normalized in the Agilent GeneSpring GX software (version 11.0) using the Agilent FE one-color scenario (mainly median normalization). Oral NFs and CAFs were obtained from tissues derived from 12 healthy people undergoing plastic surgery and 12 OSCC patiens undergoing surgical resection. RNA samples from NFs and CAFs were pooled respectively before hybridization.
Project description:Cancer-associated fibroblasts (CAFs) promote tumor progression through several mechanisms. MicroRNAs (miRNAs) play a key role in CAFs tumor-promoting properties, however, their role in CAFs from different topological areas in lung cancer progression remains unclear. This study aimed to characterize the miRNA expression profile of fibroblasts isolated from matched tumor front (F-CAFs), inner tumor (In-CAFs), and normal adjacent tissue (NFs) in lung adenocarcinoma, using microarray analysis and RT-qPCR. Proliferation and invasion assays of A549 lung cancer cells were performed in the presence of conditioned medium from F-CAFs, In-CAFs and NFs. Pathway enrichment analysis and miRNA-target networks were performed to identify tumorigenesis-related miRNAs. Both F-CAFs and In-CAFs enhanced the proliferation and invasion of A549 cells compared to NFs; however, F-CAFs showed a significantly stronger effect than In-CAFs. qPCR demonstrated three downregulated miRNAs in F-CAFs versus NFs (miR-145-3p, miR-299-3 p, miR-505-3p) and two in F-CAFs versus In-CAFs (miR-410-3p, miR-485-5p). Deregulated miRNAs showed significant association to “pathways in cancer”, “Wnt signaling pathway”, and “TGF-beta signaling pathway”, targeting tumor-promoting growth factors as IGF1 and VEGFA. Our findings suggest that deregulated miRNAs in F-CAFs, which showed the strongest effect on the invasive and proliferative capacity of A546 cancer cells, present potential predictive association with tumor-promoting properties.
Project description:Cancer-associated fibroblasts (CAFs) have been reported to support tumor progression by a variety of mechanisms. However, their role in the progression of non-small cell lung cancer (NSCLC) remains poorly defined. In addition, the extent to which specific proteins secreted by CAFs contribute directly to tumor growth is unclear. To study the role of CAFs in NSCLC, a cross-species functional characterization of mouse and human lung CAFs was performed, including gene expression analysis comparing normal mouse lung fibroblasts (NFs) and mouse lung CAFs to seek for differentially-expressed secreted proteins. Gene expression microarrays were used to identify transcriptomic changes between NFs and CAFs that may contribute to their different tumor-enhancing capacity. NFs and CAFs were grown in vitro for RNA extraction and hybridization on mouse 430_2 Affymetrix microarrays
