GSE110301
Transcriptome analysis of pathways and drug transporters after hyperosmotic stress of renal MDCK I cells
Purpose: To perform pathway analysis of MDCK I treated with hyperosmolality and to investigate how hyperosmolality affects the expression of membrane transporters. Method: MDCK I cells were treated for 24 h with medium supplemented with 200 mOsm raffinose, NaCl or urea (total of 500 mOsm). The control was regular medium (300 mOsm). RNA was isolated from 12 replicates from each condition and a cDNA library was prepared using TruSeq RNA Library Preparation Kit v2 from Illumina following the manufacturer's protocol. The sequencing was done with a HiSeq with single read and a length of 51. The alignment was done with Hisat 2, the count with FeatureCount and the statistical analysis with DESeq2. Results: The pathways the cell cycle, the cytokine-cytokine receptor interaction pathway, and the chemokine-signaling pathway were significantly activated in MDCK I cells after treatment of 200 mOsm raffinose or NaCl (total: 500 mOsm), and not 200 mOsm urea, for 24 hours. Several membrane transporters were regulated by hyperosmolality. Conclusion: Hyperosmolality has an inflammatory-like response in MDCK I cells and it also affects the expression of several membrane transporters. Overall design: Investigate the expression of membrane transporters in 200 mOsm raffinose, NaCl or urea (total: 500 mOsm) treated MDCK I cells compared to the control (300 mOsm). Performed with 12 replicates per condition.