Salvador‐Warts‐Hippo Genes Methylation status in Osteosarcoma Cell Lines

Osteosarcoma is an aggressive malignant bone tumour greatly prevalent in children and adolescent. The Salvador‐Warts‐Hippo signalling pathway is a highly conserved and is important in regulating organ size, cell proliferation and cell death. Tumorigenesis has been extensively studied along with the genetic alterations of the promoters of Salvador‐Warts‐Hippo genes which promote poor prognosis. Although, the epigenetical modifications, which present a fundamental controlling points for genetic regulations, remain inadequately clear. Such as the methylation status of CpG islands which are associated to key components genes of the hippo signalling pathway which includes MST1, MST2, LATS1, LATS2, WW45 and SAV1. We applied two pairs of primers were needed with one pair specific for methylated DNA (M) and the other for unmethylated DNA in several bone cancer cell lines including SaOS2, HT1080 and Ewing’s sarcoma A673. Combined Bisulfite Restriction Analysis (CoBRA) carried out on these cell lines and the methylation patterns were varied between osteosarcoma and Ewing’s sarcoma cells. This approach of combination techniques provides a clear note of the methylation status profile of CpG islands that governs the expression of these Salvador‐Warts‐Hippo genes in osteosarcoma cell lines. In addition, the level of expression of Salvador‐Warts‐Hippo pathway genes during demethylating agents treatment play a corresponding evidence of the biological relevance that the methylation status has on the transcriptome. The outcomes of the present study offer distinguishable means that allow the ability to differentiate between several epigenetic variations in different osteosarcoma cell lines. We conclude that Salvador‐Warts‐Hippo needs further investigation in osteosarcoma progression. Support or Funding Information Deanship of graduate studies, King Saud University, Riyadh, Saudi Arabia