Project description:This SuperSeries is composed of the following subset Series: GSE18952: Effects of IGF 1 on primary breast fibroblasts (normal and carcinoma associated) GSE18953: Effects of IGF 1 on MCF-7 breast cancer cells GSE18954: Effects of IGF 1 on CCL-171 fibroblasts Refer to individual Series
Project description:To characterize the effects of IGF 1 on primary breast fibroblasts, we cultured pre-starved primary breast fibroblasts (normal and carcinoma associated) from three patients with or without 50ng/ml IGF 1 for 24 h. We then profiled gene expression changes using Human Exonic Evidence Based Oligonucleotide (HEEBO) microarrays. After stimulation, total RNA was extracted and amplified using a modified Eberwine procedure. The amplified RNA was labeled with the fluorescent dye Cy5 and pooled with Cy3 labeled reference RNA, and then the pooled RNA was hybridized onto HEEBO microarrays. After hybridization and washing, arrays were scanned on a fluorescent microscope scanner.
Project description:To characterize the effects of IGF 1 on primary breast fibroblasts, we cultured pre-starved primary breast fibroblasts (normal and carcinoma associated) from three patients with or without 50ng/ml IGF 1 for 24 h. We then profiled gene expression changes using Human Exonic Evidence Based Oligonucleotide (HEEBO) microarrays. After stimulation, total RNA was extracted and amplified using a modified Eberwine procedure. The amplified RNA was labeled with the fluorescent dye Cy5 and pooled with Cy3 labeled reference RNA, and then the pooled RNA was hybridized onto HEEBO microarrays. After hybridization and washing, arrays were scanned on a fluorescent microscope scanner. An all pairs experiment design type is where all labeled extracts are compared to every other labeled extract.
Project description:Head and neck squamous cancer stromal fibroblasts produce growth factors influencing phenotype of normal human keratinocytes. Epithelial-mesenchymal interaction between stromal fibroblasts and cancer cells influences the functional properties of tumor epithelium including the tumor progression and spreading. We compared fibroblasts prepared from stroma of squamous cell carcinoma and normal dermal fibroblasts concerning their biological activity towards normal keratinocytes assessed by immunocytochemistry and profiling of gene activation for growth factors/cytokines by microarray chip technology. IGF-2 and BMP-4 were determined as candidate factors responsible for tumor associated fibroblast activity that influence normal epithelia. This effect was confirmed by addition of recombinant IGF-2 and BMP4 respectively to the culture medium. This hypothesis was also verified by inhibition experiments where blocking antibodies were employed in medium conditioned by cancer associated fibroblast. Presence of these growth factors was also detected in tumor samples.