Human carcinoma-associated fibroblasts (CAF): co-culture with cancer cells vs. CAF only
Ontology highlight
ABSTRACT: Gene expression profiling of CAF co-cultured with human oral squamous cell carcinoma (OSCC) cells compared with mono-cultured CAF. To identify key molecular regulators expressed by carcinoma-associated fibroblasts (CAF) that promote cancer cell invasion, microarrays were performed by comparing co-cultured OSCC cells and CAF with monoculture controls. comparison 1: CAF co-cultured with OSCC vs. mono-cultured CAF comparison 2: OSCC co-cultured with CAF vs. mono-cultured OSCC 1.7X10(5) YD-10B OSCC cells and 1.7X10(5) CAF were seeded in the upper chamber and lower chamber, respectively, of 6-transwell plates containing collagen-coated 1 micrometer pore transmembrane filters (Becton Dickinson, Franklin Lakes, NJ, USA). Monoculture control samples were generated by culturing only CAF or OSCC on the same side of the filter as in the co-culture design.
Project description:Gene expression profiling of CAF co-cultured with human oral squamous cell carcinoma (OSCC) cells compared with mono-cultured CAF. To identify key molecular regulators expressed by carcinoma-associated fibroblasts (CAF) that promote cancer cell invasion, microarrays were performed by comparing co-cultured OSCC cells and CAF with monoculture controls.
Project description:We co-cultured primary cultured CAF (CAF-P), which promotes progression more actively, and CAF (CAF-D), which have less cancer-promoting properties, with OSCC cells, and analyzed the differential secretion through antibody microarray.
Project description:The increased M-NM-1-smooth muscle-actin positive cancer-associated fibroblastic cells (CAF) in the desmoplastic stroma may relate to a more aggressive cancer and worse survival outcomes for intrahepatic cholangiocarcinoma (ICC) patients We developed a novel 3-D organotypic culture model by co-culturing M-NM-1-SMA positive CAF and cholangiocarcinoma cells in a collagen matrix. Cholangiocarcinoma cell lines were established by isolating M-NM-1-SMA positive cancer-associated fibroblastic cells (CAF) (BDEsp-TDFE4) and cholangiocarcinoma cells (BDEsp-TDEH10) from tumors arising from bile duct inoculation of spontaneously-transformed low grade malignant rat BDE1 cholangiocytes (BDEsp cells). These tumor-derived cells lines were then grown in a rat tail type I collagen gel matrix, alone or in co-culture, and their gene expression profile were compared.
Project description:The CLS1/CAF co-culture maintained the cancer stemness. This cancer stemness was lost when the CAF feeder cells were removed during passaging. To investigate the specific signaling pathways and markers of lung CSCs, we analyzed the gene expression profile of the CLS1 sphere after CLS1/CAF co-culture and compared this profile with that obtained for CLS1 cells cultured without feeder cells through different passages (CLS1 p3, p6, and p14) and CLF1 CLS1 sphere generated by co-culture with CAF and remove CAF to mimic cancer cell differentiation. Four time points of the CLS1 after co-culture with CAF have been collected for microarray analysis:CLS1-2 p.3, p.6, p.8, p.14. We tried to compare theCLS1 sphere-regulated gene expression profiles with the CLS1-2 p.3, p.6, p.8, p.14 and CLF1 to identify the possible cross talk mechanisms of CLF1 and CLS1 and the signaling pathways that maintain stem cell properties.
Project description:Cellular differentiation involves profound changes in the chromatic landscape, yet the mechanisms by which somatic cell identity is subsequently maintained remain incompletely understood. To further elucidate regulatory pathways that safeguard the somatic state, we performed two comprehensive RNAi screens targeting chromatin factors during transcription factor-mediated reprogramming of mouse fibroblasts to induced pluripotent stem cells (iPSCs). Remarkably, subunits of the chromatin assembly factor-1 (CAF-1) complex emerged as the most prominent hits from both screens, followed by modulators of lysine sumoylation and heterochromatin maintenance. Suppression of CAF-1 increased reprogramming efficiency by several orders of magnitude and facilitated iPSC formation in as little as 4 days. Mechanistically, CAF-1 suppression led to a more accessible chromatin structure at enhancer elements early during reprogramming. These changes were accompanied by a decrease in somatic heterochromatin domains, increased binding of Sox2 to pluripotency-specific targets and activation of associated genes. Notably, suppression of CAF-1 also enhanced the direct conversion of B cells into macrophages and fibroblasts into neurons. Together, our findings reveal the histone chaperone CAF-1 as a novel regulator of somatic cell identity during transcription factor-induced cell fate transitions and provide a potential strategy to modulate cellular plasticity in a regenerative setting. Gene expression analysis in CAF-1 knockdown and Renilla control during early OKSM-induced reprogramming by microarray
Project description:Molecular mechanisms of the cancer cells-carcinoma associated fibroblasts (CAF) interactions growing in vitro conditions as a co-culture. The five canine mammary cancer cell lines were cultured with CAF (isolated from canine mammary cancer) for 72hrs. Then, the cancer cells and CAFs were sorted and the gene expression analysis was conducted. The control for each co-cultured cell line was the same cell line growing as a single culture. Dye-swap experiment, each cell line growing in the co-culture conditions was compared to the same cell line growing as a single culture.
Project description:Molecular mechanisms of the cancer cells-carcinoma associated fibroblasts (CAF) interactions growing in vitro conditions as a co-culture. The five canine mammary cancer cell lines were cultured with CAF (isolated from canine mammary cancer) for 72hrs. Then, the cancer cells and CAFs were sorted and the gene expression analysis was conducted. The control for each co-cultured cell line was the same cell line growing as a single culture.
Project description:Oral cancer is the sixth leading cause of the cancer-related mortality in Taiwan. More than 90% of oral cancers are oral cavity squamous cell carcinomas (OSCCs). The high mortality rate of OSCC is ascribed to metastasis and local regional relapse of cancer cells, suggesting that identification of proteins involved in migration and chemoresistance of OSCC cells can facilitate development of useful treatment approaches for OSCC. To identify novel targets for improvement of OSCC therapy, we have comparatively profiled the proteomes of primary and relapsed OSCC tissues with iTRAQ-based mass spectrometry. Gene expression of proteins dysregulated in OSCC tissues was further surveyed with the cancer genome atlas (TCGA) database. With the analysis, we have identified proteins differentially expressed in the primary and/or relapsed OSCC tissues. Proteins up-regulated in recurrent OSCC and associated with OSCC survival have been selected for evaluation as potential therapeutic targets of relapsed OSCC.
Project description:Cancer-associated fibroblasts (CAF) as one abundant cell type of the tumor microenvironment are known to have tumor supporting abilities by promoting tumor proliferation and modulating the immunal landscape. In this study, germ cell tumor-derived CAF were molecular and epigenetically characterized to identify CAF secreted signal molecules which play a pivotal role in the GCT-CAF interaction.
Project description:Cancer cells often rely on aerobic glycolysis for metabolism, and lactylation, a newly discovered post-translational modification, significantly impacts molecular processes. This study comprehensively analyzes lactylation's role in oral squamous cell carcinoma (OSCC), providing initial insights into its impact on progression. Six pairs of OSCC tissues underwent lactylation modification sequencing. 2,765 lactylation modification sites on 1,033 proteins were identified in OSCC, indicating its widespread presence. These modifications influenced metabolic activities, molecular synthesis, and transport.