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. 2020 Jun 9:13:5271-5281.
doi: 10.2147/OTT.S227331. eCollection 2020.

Destruction of Neutrophil Extracellular Traps Promotes the Apoptosis and Inhibits the Invasion of Gastric Cancer Cells by Regulating the Expression of Bcl-2, Bax and NF-κB

Rong Li  1 , Xiaoming Zou  1 , Tong Zhu  1 , Haiyan Xu  2 , Xiaolin Li  1 , Lei Zhu  1
Affiliations

Destruction of Neutrophil Extracellular Traps Promotes the Apoptosis and Inhibits the Invasion of Gastric Cancer Cells by Regulating the Expression of Bcl-2, Bax and NF-κB

Rong Li et al. Onco Targets Ther. .

Abstract

Introduction: This study aimed to investigate the effects of Neutrophil extracellular traps (NETs) destruction on the apoptosis and invasion of gastric cancer cells and the involved mechanisms.

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).

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.

Keywords: cell apoptosis; gastric cancer; neutrophil; neutrophil extracellular traps.

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Conflict of interest statement

The authors report no conflicts of interest in this work.

Figures

Figure 1
Figure 1
The expression of NETs (SPINK5/LEKTI) (A) and Cit Histone H3 (B) which was detected by immunofluorescent analysis and Western blot, respectively. *P < 0.05 vs Control.
Figure 2
Figure 2
The apoptosis of three gastric cancer cells (BGC-823, SGC7901 and MKN28) in various groups which was examined by flow cytometry. Three gastric cancer cells were co-cultured with neutrophils respectively. *P < 0.05 vs Control.
Figure 3
Figure 3
The invasion of three gastric cancer cells (BGC-823, SGC7901 and MKN28) in various groups which was evaluated by Transwell assay. *P < 0.05 vs Control.
Figure 4
Figure 4
The mRNA levels of NF-κB p65, Bcl-2 and Bax in three gastric cancer cells (BGC-823, SGC7901 and MKN28) of various groups which were evaluated by RT-PCR. *P < 0.05 vs Control.
Figure 5
Figure 5
The protein levels of NF-κB p65, Bcl-2 and Bax in three gastric cancer cells (BGC-823, SGC7901 and MKN28) of various groups which were evaluated by immunofluorescent analysis. *P < 0.05 vs Control.
Figure 6
Figure 6
The protein levels of NF-κB p65, Bcl-2 and Bax in three gastric cancer cells (BGC-823, SGC7901 and MKN28) of various groups which were evaluated by Western blot. *P < 0.05 vs Control.
See this image and copyright information in PMC

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