Expression profiles of cell populations purified from human CRC (3 ways)

ABSTRACT: The survival of isolated metastatic cells and expansion into macroscopic tumour has been recognized as a limiting step for metastasis formation in several cancer types yet the determinants of this process remain largely uncharacterized. In colorectal cancer (CRC), we identify a transcriptional programme in tumour-associated stromal cells, which is intimately linked to a high risk of developing recurrent disease after therapy. A large proportion of CRCs display mutational inactivation of the TGF-beta pathway but paradoxically they are characterized by high TGF-beta production. In these tumours, TGF-beta instructs a transcriptional programme in stromal cells, which confers a high risk of developing metastatic disease. We purified by FACS [CD45(+),Epcam(-)], [CD45(-) Epcam(+)] and [CD45(-) Epcam(-)] cell populations from fresh CRC samples and assessed their gene expression profiles Freshly obtained tumors from CRC patients (n=8) treated at Hospital del Mar (Barcelona, Spain) were minced and incubated with 0.1% Hyaluronidase and 0.1% Collagenase 1A. Pieces were then homogenized. Enzymatic reaction was stopped by adding 10% FBS and single cells were collected by sequential filtering. Cells were resuspended in Ammonium Chloride (0.15M) to lyse erythrocytes. Cells were stained with anti-hEpcam/TROP1-APC and anti-CD45-PE conjugated antibody. Dead cells were labeled with Propidium Iodide. Fluorescence Activated Cell Sorting (FACS) was used to separate cells.

ORGANISM(S): Homo sapiens

SUBMITTER: Alexandre CALON 

PROVIDER: E-GEOD-39395 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress