The effects of Epigallocatechin-3-gallate and Dabrafenib combination on apoptotic and gene expression in anaplastic throid cancer cells
Project no. 20389 · Status: Kapanmış · 2019
Summary
Anaplastic Thyroid Cancer which is responsible for 40 % of throid cancer death occur rare %2 ratio. ATC molecular pathogenesis which are resistant to chemotherapeutics still unclear. BRAF V600E gene mutation leads to 45% of the ATC patients with poor prognosis. Also ıt is associated with epigenetic modifications which is abnormal DNA methylation in thyroid cancers. Dabrafenib (Dab) is a drug that inhibits MAPK signal pathway. Epigallocathesin-3-gallat (EGCG) is a polyphenol compound of grean tea which is still under investigation on the role of cancer. Its known epigentic diet as DNA methyltransferase inhibitör. In thesis, we aimed that increasing anti-tumor effect of Dab. combination with EGCG and inducing apoptosis by possible synergistic activity and determination of epigenetic related gene expression. Treatments applied on BRAF V600E/mut SW 1736 cell line as a ATC model in vitro. The experiment consist of SW 1736 cell viability with trypan blue staining test and the stotoxic effects of Dab and EGCG with WST- 8 cell count assay following apoptotic effect was determined using Annexin V-FITC method. Gen expression changes of 84 genes associated with epigenetic mechanisms were analyzed by real-time RT-qPCR. Dab. and EGCG combination which was first applied on SW 1736 cells significantly reduced cell viability due to concentration and to induce apoptosis 33 fold with additive effect together. The determined IC50 doses are Dab 6.3 µM and EGCG 22.5 µM for 48 hours. Significant changes were defined on 24 gene expression after drugs treatment on sw 1736 cell line which committed to the literature as a guide for ATC advanced molecular researches. We suggest that, the combination of Dab and EGCG following in vivo phase studies will contribute to clinically on ATC therapy.