Total RNA was extracted from frozen tissue of PTCLs and reactive hyperplasia, as well as bone marrow blasts of T-ALL, using TRIzol reagent (Invitrogen). cDNA was synthesized using PrimeScript RT reagent Kits with gDNA Eraser (TaKaRa, CA, USA) following the manufacturer’s instructions. Real-time polymerase chain reaction (PCR) was performed using SYBR Premix Ex Taq™ II (TaKaRa) on ABI Prism 7500 (Applied Biosystems, Bedford, MA, USA). The relative gene expression levels were calculated using the SDS2.4 software. The primer sequences were as follows: c-FLIPL forward, 5′-ATTGCATTGGCAATGAGACAGAGC-3′; reverse, 5′-TCGGTGCTCGGGCATACAGG-3′, c-FLIPS forward, 5′-ACCCTCACCTTGTTTCGGACTAT-3′; reverse, 5′-TGAGGACACATCAGATTTATCCAAA-3′, TRAIL forward, 5′-TCAGCACTTCAGGATGATGG-3′; reverse, 5′-CACCAGCTGTTTGGTTCTCA-3′, DR5 forward, 5′-TGACGGGGAAGAGGAACTGA-3′; reverse, 5′-GGCTTTGACCATTTGGATTTGA-3′, GAPDH forward, 5′-GAAGGTGAAGGTCGGAGTC-3′; reverse, 5′-GAAGATGGTGATGGGATTTC-3′.
Real-time PCR of NF-κB signaling pathway was performed using the RT2 profiler PCR Array-Human NF-κB signaling pathway (QIAGEN Sciences, Frederick, MD, USA). GAPDH was used as the endogenous control and Jurkat cells for calibration. A relative quantification was calculated using the ΔΔCT method.
Real-time PCR of NF-κB signaling pathway was performed using the RT2 profiler PCR Array-Human NF-κB signaling pathway (QIAGEN Sciences, Frederick, MD, USA). GAPDH was used as the endogenous control and Jurkat cells for calibration. A relative quantification was calculated using the ΔΔCT method.