Transcriptional differences in suspended breast cancer cells exposed to hypoxia.

We report the transcriptional differences of suspended MCF-7 and MDA-MB-231 breast cancer cells after exposure to hypoxia. MCF-7 cells (1*106 cells/well) and MDA-MB-231 cells (1*106 cells/well) were suspended in high-glucose DMEM containing 10% fetal bovine serum and 1% penicillin/streptomycin at 37°C, 5%CO2 and seeded in individual wells of ultra-low attachment 6-well culture plates (Corning® Costar®, CLS3471). To simulate a hypoxic environment, we placed suspension-cultured tumor cells in a sealed Rectangular jar(MITSUBISHI GAS CHEMICAL, JAR-25) with hypoxia indicator with AnaeroPack™- Anaero(MITSUBISHI GAS CHEMICAL) and incubated for 6hrs hrs. RNA was extracted and submitted for library preparation, RNA sequencing and preliminary analysis.