BioProject

TGF-beta is one of the key cytokines implicated in various disease processes including cancer. More...

TGF-beta is one of the key cytokines implicated in various disease processes including cancer. TGF-beta inhibits growth and promotes apoptosis in normal epithelial cells and in contrast, acts as a pro-tumour cytokine by promoting tumour angiogenesis, immune-escape and metastasis. It is not clear if various actions of TGF-beta on normal and tumour cells are due to differential gene regulations. Hence we studied the regulation of gene expression by TGF-beta in normal and cancer cells. Results: Using human 19 K cDNA microarrays, we show that 1757 genes are exclusively regulated in A549 cells in contrast to 733 genes exclusively regulated in HPL1D cells. In addition, 267 genes are commonly regulated in both the cell lines. Semi quantitative and real-time qRT-PCR analysis of some genes agrees with the microarray data. Conclusion: These data suggest major differences with respect to TGF-beta mediated gene regulation in normal and transformed cells. Keywords: cell type comparison in response to TGF-beta Overall design: To compare the genes regulated by TGF-beta in non transformed and transformed cells, we chose an immortalized lung epithelial cell line, HPL1D and a lung adenocarcinoma cell line, A549. Cells were plated and propagated in designated media and TGF-beta treatments were carried out in serum free conditions at various time intervals, 1hr, 4hr and 12hr. The RNA was extracted and expression profiling was performed using microarray experiments on 19K cDNA arrays after labeling the cDNAs with Cy3 and Cy5 dyes for normal and tumor cell RNAs respectively. The image and data analysis was performed using Quantarray (P.E. Life Sciences, USA) and Avadis (Strand Life Sci. India) softwares. The results are computed as log2 transformed fold changes between treated and untreated respective cell lines. The data of regulated genes of each cell line was compared with the other cell line and exclusively and commonly regulated genes have been determined. Less...