Conclusion
NETs destruction promoted the apoptosis and inhibited the invasion of gastric cancer cells by regulating the expression of Bcl-2, Bax and NF-κB.
Methods
Primary human neutrophils were isolated and co-cultured with three gastric cancer cells (BGC-823, SGC7901 and MKN28), and phorbol-12-myristate-13-acetate was added to generate NETs. Expression of NETs (SPINK5/LEKTI) and Cit Histone H3 were examined by immunofluorescent analysis and Western blot. Cancer cells were then divided into five groups: Control, NETs, Neutrophil, Amidine and DNase I. Cell apoptosis and invasion were examined by Transwell assay and flow cytometry, respectively. Expression of NF-κB p65, Bcl-2 and Bax was determined by RT-PCR, immunofluorescent analysis and Western blot.
Results
The expression of NETs (SPINK5/LEKTI) and Cit Histone H3 after co-culture increased significantly (P < 0.05), suggesting that gastric cancer cells could promote NETs generation. The Control, NETs and Neutrophil groups exhibited similar apoptosis and invasion of gastric cancer cells and similar mRNA and protein levels of NF-κB p65, Bcl-2 and Bax. However, compared with the Control group, the apoptosis and invasion of gastric cancer cells in both Amidine and DNase I groups were enhanced and weakened, respectively (P < 0.05). Moreover, both Amidine and DNase I groups showed much higher mRNA and protein levels of NF-κB p65 and Bax and lower mRNA and protein levels of Bcl-2 than the Control group (P < 0.05).