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. 2019 Jul 10;11:6379–6396. doi: 10.2147/CMAR.S201610

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© 2019 Jing et al.

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Knockdown of DNMT3a promotes apoptosis in PANC-1 cells. (A) The cell apoptosis was increased as detected by flow cytometry in PANC-1 cells after siRNA DNMT3a treatment for 72 hrs. The apoptosis was reduced in cells with both DNMT3a and caspase-8 knockdown. (B) The apoptosis rates of PANC-1 cells treated with siRNA DNMT3a and siRNA CASP8 (annexin V positive cells, %). *P<0.05 vs. parental cells with siRNA NC treatment, **P<0.01 vs cells with siRNA normal control NC treatment. (C) Western blotting analysis showed that the expression of apoptosis-related proteins (cleavage of caspase-3, caspase-8, cleavage of caspase-8, and PARP) was elevated in PANC-1 cells with siRNA DNMT3a transfection. (D) Real-time qPCR showed elevated mRNA levels of CASP8 in DNMT3a knockdown PANC-1 cells. **P<0.01 vs cells with siRNA normal control NC treatment. (E) No expression change was detected in the expressions of exogenous apoptosis-related proteins FADD, TNFR1, DR4, and DR5 in siRNA DNMT3a treated PDAC cells. (F) Co-immunoprecipitation assay revealed that DNMT3a down-regulation increased the combination of FADD and caspase-8, and enhanced the cleavage of caspase-8 accordingly.

Abbreviations: DNMT, DNA methyltransferase; PDAC, pancreatic ductal adenocarcinoma.