Evaluating Stemness Markers in Brain Tumors

Gene expression was assessed using real-time RT-PCR and data expressed relative to the housekeeping gene, HPRT, as previously reported [20 (link)]. The expression of CD133, VEGF and OCT4, which are commonly used to define the CSC-like population in brain tumours, were detected using SYBR Green (Eurogentec) and calculated using the 2^−∆∆Ct method. The primer sequences used were: CD133 forward: 5′-CAATCTCCCTGTTGGTGATTTG-3′; and CD133 reverse: 5′-ATCACCAGGTAAGAACCCGGA-3′; VEGF forward: 5′-CCAAGTGGTCCCAGGCTGCA-3′; reverse: 5′-TGGATGGCAGTAGCTGCGCT-3′. OCT4 forward: 5′-GTTGGAGAAGGTGGAACCAA-3′; and OCT4 reverse: 5′-CTCCTTCTGCAGGGCTTTC-3′.