Project description:The unfolded protein response (UPR) is activated in response to hypoxia-induced stress such as in the tumor microenvironment. This study examined the role of CREB3L1 (cAMP-responsive element-binding protein 3-like protein 1), a member of the UPR, in breast cancer development and metastasis. Initial experiments identified the loss of CREB3L1 expression in metastatic breast cancer cell lines compared to low- or non-metastatic cell lines. When metastatic cells were transfected with CREB3L1 they demonstrated reduced invasion and migration in vitro, as well as a significantly decreased ability to survive under non-adherent or hypoxic conditions. Interestingly, in an in vivo rat mammary tumor model, CREB3L1 expressing cells not only failed to form metastases compared to CREB3L1 null cells but regression of the primary tumors was seen in 70% of the animals as a result of impaired angiogenesis. Microarray and ChIP on Chip analyses identified changes in the expression of many genes involved in cancer development and metastasis, including a decrease in those involved in angiogenesis. These data suggest that CREB3L1 plays an important role in suppressing tumorgenesis and loss of expression is required for the development of a metastatic phenotype. CREB3L1 is a member of the unfolded protein response family of proteins. CREB3L1 expression is lost from metastatic breast cancer cells. We wanted to determine the promoters of genes that CREB3L1 bound to. Idenification of promoters to which CREB3L1 is bound following ChIP with a HA antibody in MDA-MB-435 cells.
Project description:MDA-MB-435 breast cancer cells were treated with 2ME2 (2 µM) or vehicle alone. RNA was extracted and genomic profiling was performed using 22k Agilent microarrays. Experiment Overall Design: MDA-MB-435 breast cancer cells were treated with 2ME2 (2 µM) or vehicle alone. RNA was extracted and genomic profiling was performed using 22k Agilent microarrays.
Project description:This experiment is designed to screen miRNAs that are deregulated during breast cancer metastasis. Comparatively analyzing miRNAs in parental MDA-MB-435 cells and cells obtained from its lung metastases, 23 miRNAs expressed differentially, among which 12 were elevated and 11 were down-regulated. Total RNA were extracted from parental MDA-MB-435 cells and cells obtained from its lung metastases after 30 days inoculation. Two biological replications for each treatment.
Project description:Expression of the transcription factor CEBPD is induced at early stages of the endoplasmic reticulum (ER) stress response. In order to identify the genes modulated by CEBPD during the ER stress response, we transiently silenced CEBPD expression in MDA-MB-435 melanoma cells and isolated mRNA at 6 h of treatment with Thapsigargin or DMSO control. As control, cells were transfected with two different control siRNA oligonucleotides. Results provide insights into which genes are modulated by CEBPD and/or Thapsigargin in MDA-MB-435 cell
Project description:The objective of this study was to determine the gene expression changes mediated by the alpha6beta4 integrin using MDA-MB-435 breast carcinoma cell line under normal culturing conditions (10% FCS in DMEM). Experiment Overall Design: Comparison of clones of MDA-MB-435 cells transfected with the integrin beta4 subunit (which results in cell surface expression of the integrin apha6beta4 integrin; clones 3A7 and 5B3) to those transfected with vector (mock; 6D2 and 6D7) only.
