Expression profiling of human neuroblastoma cells SK-N-SH after treatment with low dose of 2,3,7,8- tetrachlorodibenzo-p-dioxin

Exposure to dioxins has been associated with impaired development of the nervous system and losses of brain functions. However, the precise cellular events and related molecular basis underlining these adverse effects has not been fully understood. We aimed to reveal global alterations of mRNA expression and potential cellular events by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure. Messenger RNA microarray analysis was employed on cultured human neuroblastoma cells SK-N-SH exposed to 10-10 M TCDD.Total RNA was extracted from cell samples using Trizol/Chloroform method, and then purified with magnetic beads of Agencourt Ampure (Beckman Coulter, Brea, CA, USA). Target preparation for microarray processing was carried out according to the GeneChip® WT PLUS Reagent Kit. In each preparation, 500 ng total RNA was used for a double-round of cDNA synthesis. After fragmentation of 2nd-cycle single-stranded cDNA, sample was labeled with biotin by terminal deoxynucleotidyl transferase (TdT). Then sample was hybridized to a GeneChip® Human Transcriptome Array 2.0 (HTA 2.0, Affymetrix) with 44699 annotated genes.