Transcriptional profiling of senescent A549 cells

Telomere dysfunction is the most common identifiable cause of pulmonary fibrosis. Cellular senescence of epithlial cells is thought to play a signfiicant role in disease pathogensis. We established a model of cellular senescence using a human alveolar epithelilial-like cell lines (A549) and profiled the transcriptional changes that occur in response to telomere dyfunction. Telomere dysfunction was induced by expressing a dominant-negative form TRF2, the shelterin binding protein. RNA-seq was performed on 12 samples from 4 groups (3 biologic replicates each). The four groups are as follows; 1) A549 cells. 2) A549 cells cultured with 2 microgram per mL doxicycline for 10 days. 3) A549 cells stably expressing a doxicylcine-inducible TRF2-DN protein that induces telomere dysfunction and cellular scenescence when expressed. And 4) TRF2-DN cells cultured with 2 micrograms per mL doxicycline for 10 days. RNA was isolated using RNAeasy kits (Qiagen) and total RNA was sent to Novagen for paired end RNA-sequencing. At least 20 million reades were sequenced per sample.