Apoptosis of pancreatic carcinoma cell strain PANC-1 induced by knockdown of NF-κB p65 expression with RNA interference

Objective To observe the apoptotic effects of RNAi-mediated NF-κB p65 gene on silencing pancreatic carcinoma cell strain PANC-1. Methods Chemically synthesized small interference RNA (siRNA) directed against human NF-κB p65 was transfected into pancreatic carcinoma cell PANC-1 using cationic liposome LipofectamineTM 2000 as the transfecting agent. The expression of NF-κB p65 gene was detected by RT-PCR technique. The DNA binding activity of NF-κB was detected by the chemicon non-radioactive NF-κB p65 transcription factor assay kit. Nuclear DNA staining by Hoechst 33258, flow cytometry, transmission electron microscope was used to ascertain whether cell apoptosis could be induced by down-regulating the expression of p65 gene in PANC-1 cell. Results The results of RT-PCR indicated that chemically synthesized siRNA directed against human NF-κB p65 could knock down the transcription and expression of NF-κB p65 gene. The difference between the RelA siRNA group and control groups was significant (P0.01). After transfection, the DNA binding activity of NF-κB p65 in RelA siRNA group was much lower than that of the control groups (P0.05). Nuclear DNA staining by Hoechst 33258, flow cytometry, transmission electron microscopy showed that knockdown of p65 expression by RNAi induced apoptosis in PANC-1 cell. Conclusion The study provides basis for researching the function of NF-κB p65 gene, indicating down-regulating the expression of p65 gene by RNAi may induce apoptosis in pancreatic carcinoma cell strain PANC-1.