GATA1 Promotes Gemcitabine Resistance in Pancreatic Cancer through Antiapoptotic Pathway
Figure 3
Screening for target genes in GATA1 mediated gemcitabine resistance. (a) Schematic representation of the protocol to obtain gemcitabine-resistant PDAC cell line (Gem-R). Red and blue colors of cells represent gemcitabine resistance and sensitivity, respectively. Gem-R cells were stably passaged for more than 10 times after exposed to an increasing concentration of gemcitabine for over 3 months. (b) Cell viability assays of CFPAC-1 and CFPAC-1 Gem-R cells treated with a range of concentration of gemcitabine for 48 h. (c) qRT-PCR analysis of relative mRNA expression in CFPAC-1 versus CFPAC-1 Gem-R cells of genes related to proliferation and drug resistance. (d) qRT-PCR analysis of relative mRNA expression in CFPAC-1 cells transiently transfected with empty vector or GATA1. The genes from (c) were used for qRT-PCR. (e) Elevated expression of GATA1 and Bcl-XL was detected in CFPAC-1 Gem-R by Western blot analysis. (f) Western blot analysis of Bcl-XL and GATA1 in CFPAC-1 cells transiently transfected with empty vector or GATA1, control siRNA, or GATA1 siRNA. β-actin was used as a loading control. (g) Cell viability curves of CFPAC-1-Gem-R cells transfected with GATA1 siRNA or Bcl-XL siRNA. The cells were exposed to a range of concentration of gemcitabine for 48 h before CCK8 assay. The knockdown effects of siRNAs were confirmed by Western blot analysis, with β-actin as a loading control. All data shown are means ± SD of three independent experiments with triplicate each, ; .