A232: Methylation of Salvador-Warts-Hippo pathway in Osteosarcoma Cells
1UNIVERSIT OF SOUTHAMPTON, SOUTHAMPTON, UK,2KING SAUD UNIVERSITY, RIYADH, Saudi Arabia
DNA methylation are key epigenetic factors regulating gene expression and genomic stability. Salvador-Warts-Hippo signalling pathway is an emerging growth control and tumour suppressor pathway that regulates cell proliferation. The Hippo pathway is responsible for cell proliferation and organ size control in mammalian systems. The components of the pathway are two kinases and their adaptor proteins which inhibit the transcription co-activator YAP by phosphorylation. When the pathway is inactive, activated YAP is translocated to the nucleus where it cooperates with TEAD transcription factor and promotes expression of genes that regulate cell proliferation and apoptosis.
We applied methylation specific PCR (MSP) is a bisulfite conversion based PCR technique for the study of DNA CpG methylation. Two pairs of primers are needed with one pair specific for methylated DNA (M) and the other for unmethylated DNA (U).
The methylation of this pathway was varied between these cells and methylation frequency was detected in number of these cells. The preliminary finding shows that there are number of Hippo pathway genes were differentially methylated in these cells. In MST1 and MST2 analysis, SaOS and U205 cells showed methylation pattern, respectively. In case of SAV1, non of osteosarcoma cells showed any methylation status. In LATS1 methylation detection, only U205 cells showed methylation pattern and non of osteosarcoma cells showed any LATS2 methylation.
In conclusion, hippo pathway ippo pathway observed epigenetic variations in different osteosarcoma cell lines within the same cancer type which lead us to investigate the mechanism of these genes in Osteosarcoma cell lines.