INVESTIGATING CUDC-101 EFFECT AND MECHANISM OF ACTION AGAINST GLIOBLASTOMA IN VITRO

Abstract

AIMS Glioblastoma (GB) has complex pathophysiology, difficult treatment and resultant poor prognosis. Its median survival rate is approximately 15 months. The aggressive nature of GB results in rapid disease progression in 60-70% patients often within 12 weeks. Limited treatment options include maximal safe surgical resection, followed by radiotherapy and temozolomide (TMZ). The blood brain barrier restricts chemotherapeutic entry and increases dosage leading to increased side effects and decreased treatment tolerance. Furthermore, significant disease heterogeneity means reoccurrence is expected in most cases, for which there are no standard treatment routes. This coupled with inefficient treatment with temozolomide (TMZ) due to the blood brain barrier, highlights the requirement for novel treatments. METHOD Histone deacetylases (HDAC) and endothelial growth factor receptor (EGFR) are mutated and upregulated in GB allowing tumour infiltration and proliferation. CUDC-101 is known to target both HDAC and EGFR in other cancers making it a promising therapeutic to trial in GB. Cell viability of U251, T98G and U87MG human cells was measured post-CUDC-101 administration at 24, 48 and 72hrs. SVGp19 embryonic non-cancerous human glial cells were used as a control to establish CUDC-101 preliminary specificity. Targeting another major hallmark of GB, wound healing assays were performed to assess CUDC-101 capacity to inhibit migration. In addition to this, long-term cellular resistance was investigated through clonogenic assays. Western blotting was then used to identify non-/treated expression levels of activated EGFR and acetylated histone H3. Expression levels in non/-treated cells were then visualised with fluorescent microscopy. In addition, flow cytometry was utilised to observe cell cycle changes. RESULTS Overall, results indicate CUDC-101 has a dose-dependent effect on cell viability (long-term and short-term) and migration. Additionally, fluorescent images show treatment does increase histone acetylation and decrease total EGFR. CONCLUSION Future work will solidify these datasets and show changes in activated EGFR and acetylated histone H3 using western blotting.