Targeting metabolic pathways to radiosensitize pancreatic cancer (Human xenograft)
Mice were kept in pathogen-free condition under 12 hr light-dark cycles and room temperature between 20-25C and appropriate humidity (30-70%).Mice were orthotopically implanted with Luciferace tageed CD18/HPAF pancreatic cancer cells into the pancraes of athymic nude mice. Mice were randomized into four groups, and each group consists of 5 mice. Group 1 mice were kept as control (no IR treatment), whereas group 2 and group 3 mice received fractionated irradiation dose of approximately 7 Gy once a day for five days or Zol alone. Group 4 mice received both radiation and pre-treated with Zol. The implanted xenografts were irradiated using a linear accelerator (7 Gy of 5 fractions(SBRT)) located in the Department of Radiation Oncology at UNMC. During the sacrifice a part of xenograft tissue was flash-frozen and total RNA was isolated using Qiagen RNAeasy RNA isolation kit and as per manufacure's protocol. RNA quality of each samples were checked using Advanaced Analytical fragment analyzer. Samples with RNA integrity number 9 will be further considered for cDNA preperation and transcriptomic analysis. Library was prepared using Illumina Tru seqTM RNA ample preperation version 2 kit.