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. 2019 Feb 1;10(10):1056–1069. doi: 10.18632/oncotarget.26621

Figure 4. Src kinase regulates E-cadherin expression through the transcription factor Slug.

Figure 4

To verify whether targeting Src kinase can affect E-cadherin and Slug expression at mRNA level, RT-PCR was performed. PDAC cells were treated with DST in a dose-dependent manner (0-1000 nmol/L) for 12 h. RT-PCR analyses were performed to verify whether E-cadherin and Slug regulation by Src kinase inhibition occurs at the transcriptional level in drug-sensitive (A) and resistant (B) PDAC cells. Relative gene expression was calculated by normalizing each treatment value to corresponding GAPDH signal intensity, then reported relative to control signal intensity. Luciferase reporter assay for E-cadherin promoter activity in response to DST treatment (0-1000 nmol/L) in drug-sensitive (C) and resistant (D) PDAC cell lines was performed to determine if inhibition of Src kinase acts to enhance E-cadherin transcription in drug-sensitive cells. (E) To confirm Slug is essential for DST-mediated E-cadherin expression in drug-sensitive PDAC cells, BxPC3 cells were transfected with short-hairpin RNA (shRNA) to Slug and treated with DST (100 nmol/L). qPCR was performed to analyze relative expression levels of E-cadherin and Slug in response to shRNA knockdown and/or DST treatment. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns not significant.