Project description:To understand the growth inhibition of peptidylarginine deiminase 2 (PAD2) and small molecule 1B8, colon cancer SW480 cells were treated with compound 1B8 for 24 hours, SW480 cells were transfected with humanPAD2 vector for 24 hours. Cells were harvested and total RNA was extracted using TRIZol method followed by Agilent DNA microarray analysis. To profile the effected genes by compound 1B8, SW480 cells were treated with DMSO (control) and 10uM 1B8 in triplicates in 6-well plate, cells were harvested 24 hours later. To compare the effected genes by compound 1B8 and PAD2 overexpression, SW480 cells were transfected with PAD2 and pIRES2-EGFP empty vector in triplicates in 6 well plates, cells were harvested in 24 hours. All the cells were harvested and total RNA was extracted using TRIZol method followed by Agilent DNA microarray analysis.
Project description:Transcriptional profiling of human colon cancer SW480 cells comparing control untreated SW480 cells with cells stably transfected with LRP16 treated with or without etoposide (50 μM) for the indicated periods (0,1hour, 3hours).An exploratory microarray analysis was performed with mRNA extracted from clutured SW480 cells transfected with LRP16 or control plasmid that were treated with or without etoposide. Total RNA of colon cancer cells stably transfected with vector control and LRP16 treated with or without etoposide (50 μM) for the indicated periods was isolated and purified using RNeasy Kit (Qiagen, Hilden, Germany). Integrity of RNA was assessed by using an Agilent BioAnalyser 2100 (Agilent Technologies).