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中性粒细胞胞外诱捕网的破坏通过调节Bcl-2、Bax和NF-κB的表达促进胃癌细胞凋亡并抑制其侵袭。

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.

作者信息

Li Rong, Zou Xiaoming, Zhu Tong, Xu Haiyan, Li Xiaolin, Zhu Lei

机构信息

Department of General Surgery, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150000, People's Republic of China.

Department of Medicine, Central Hospital of Prison Administration Bureau of Heilongjiang Province, Harbin, Heilongjiang 150000, People's Republic of China.

出版信息

Onco Targets Ther. 2020 Jun 9;13:5271-5281. doi: 10.2147/OTT.S227331. eCollection 2020.

DOI:10.2147/OTT.S227331
PMID:32606746
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7293391/
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.

摘要

引言

本研究旨在探讨中性粒细胞胞外陷阱（NETs）破坏对胃癌细胞凋亡和侵袭的影响及其相关机制。

方法

分离原代人中性粒细胞并与三种胃癌细胞（BGC-823、SGC7901和MKN28）共培养，加入佛波醇-12-肉豆蔻酸酯-13-乙酸酯以生成NETs。通过免疫荧光分析和蛋白质印迹法检测NETs（丝氨酸蛋白酶抑制剂Kazal型5/淋巴上皮Kazal型抑制剂）和瓜氨酸化组蛋白H3的表达。然后将癌细胞分为五组：对照组、NETs组、中性粒细胞组、脒基组和脱氧核糖核酸酶I组。分别通过Transwell实验和流式细胞术检测细胞凋亡和侵袭情况。通过逆转录聚合酶链反应、免疫荧光分析和蛋白质印迹法测定核因子κB p65、B细胞淋巴瘤/白血病-2（Bcl-2）和Bax的表达。

结果

共培养后NETs（丝氨酸蛋白酶抑制剂Kazal型5/淋巴上皮Kazal型抑制剂）和瓜氨酸化组蛋白H3的表达显著增加（P<0.05），表明胃癌细胞可促进NETs生成。对照组、NETs组和中性粒细胞组的胃癌细胞凋亡和侵袭情况以及核因子κB p65、Bcl-2和Bax的mRNA和蛋白水平相似。然而，与对照组相比，脒基组和脱氧核糖核酸酶I组的胃癌细胞凋亡分别增强和减弱（P<0.05）。此外，与对照组相比，脒基组和脱氧核糖核酸酶I组的核因子κB p65和Bax的mRNA和蛋白水平均显著升高，而Bcl-2的mRNA和蛋白水平均显著降低（P<0.05）。

结论

NETs破坏通过调节Bcl-2、Bax和核因子κB的表达促进胃癌细胞凋亡并抑制其侵袭。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/68ab/7293391/6d03bc6a0627/OTT-13-5271-g0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/68ab/7293391/b3ee655d72f5/OTT-13-5271-g0001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/68ab/7293391/6d03bc6a0627/OTT-13-5271-g0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/68ab/7293391/b3ee655d72f5/OTT-13-5271-g0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/68ab/7293391/7014aabd1695/OTT-13-5271-g0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/68ab/7293391/f88e2797ae4e/OTT-13-5271-g0003.jpg
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