Expression data of TPC-1 cells with SMAD3 knockdown

The human SMAD3 gene consists of 9 exons, has 4 isoforms, is located in chromosome 15 and measures 129,339bp in size. Loss of SMAD3 expression was found in gastric cancer and T cell acute lymphoblastic leukemia. Recent genome-wide association studies (GWAS) for thyroid cancer suceptibility discovered 15q22 as one of the significantly risky loci. Two tag SNPs of this locus are located in the intronic region of SMAD3 gene. In order to reveal the regulation gene network of SMAD3 in thyroid cells, microarray analysis was performed in TPC-1 cell line treated with si-SMAD3. TPC-1 cells were seeded in 6-well-plate and grown to ~85% confluence at the time of transfection. 25pmol si-SMAD3 or si-Control were transfected into the cells using 4.5ul RNAiMAX lipofectamine (Thermo Fisher SCIENTIFIC). After transfection for 24h, the cells were harvested and total RNA was extracted by using Trizol reagent (Thermo Fisher SCIENTIFIC) then treated by DNase-I (Ambion) to eliminate DNA contamination. Experiments described above were performed at three different time points in order to obtain three replicates for expression analysis.