PDF(Pubmed)
Abstract：
Neutrophils are one of the most predominant infiltrating leukocytes in lung cancer tissues and are associated with lung cancer progression. How neutrophils promote lung cancer progression, however, has not been established.
Kaplan-Meier plotter online analysis and tissue immunohistochemistry were used to determine the relationship between neutrophils and overall survival in lung cancer patients. The effect of neutrophils on lung cancer was determined using the Transwell migration assay, a proliferation assay, and a murine tumor model. Gene knockdown was used to determine poly ADP-ribose polymerase (PARP)-1 function in lung cancer-educated neutrophils. Western blot analysis and gelatin zymography were used to demonstrate the correlation between PARP-1 and matrix metallopeptidase 9 (MMP-9). Immunoprecipitation coupled to mass spectrometry (IP/MS) was used to identify the proteins interacting with PARP-1. Co-immunoprecipitation (Co-IP) was used to confirm that PARP-1 interacts with arachidonate 5-lipooxygenase (ALOX5). Neutrophil PARP-1 blockage by AG14361 rescued neutrophil-promoted lung cancer progression.
An increased number of infiltrating neutrophils was negatively associated with overall survival in lung cancer patients (P < 0.001). Neutrophil activation promoted lung cancer cell invasion, migration, and proliferation in vitro, and murine lung cancer growth in vivo. Mechanistically, PARP-1 was shown to be involved in lung cancer cell-induced neutrophil activation to increase MMP-9 expression through interacting and stabilizing ALOX5 by post-translational protein modification (PARylation). Blocking PARP-1 by gene knockdown or AG14361 significantly decreased ALOX5 expression and MMP-9 production, and eliminated neutrophil-mediated lung cancer cell invasion and in vivo tumor growth.
We identified a novel mechanism by which PARP-1 mediates lung cancer cell-induced neutrophil activation and PARylates ALOX5 to regulate MMP-9 expression, which exacerbates lung cancer progression.
Kaplan-Meier plotter online analysis and tissue immunohistochemistry were used to determine the relationship between neutrophils and overall survival in lung cancer patients. The effect of neutrophils on lung cancer was determined using the Transwell migration assay, a proliferation assay, and a murine tumor model. Gene knockdown was used to determine poly ADP-ribose polymerase (PARP)-1 function in lung cancer-educated neutrophils. Western blot analysis and gelatin zymography were used to demonstrate the correlation between PARP-1 and matrix metallopeptidase 9 (MMP-9). Immunoprecipitation coupled to mass spectrometry (IP/MS) was used to identify the proteins interacting with PARP-1. Co-immunoprecipitation (Co-IP) was used to confirm that PARP-1 interacts with arachidonate 5-lipooxygenase (ALOX5). Neutrophil PARP-1 blockage by AG14361 rescued neutrophil-promoted lung cancer progression.
An increased number of infiltrating neutrophils was negatively associated with overall survival in lung cancer patients (P < 0.001). Neutrophil activation promoted lung cancer cell invasion, migration, and proliferation in vitro, and murine lung cancer growth in vivo. Mechanistically, PARP-1 was shown to be involved in lung cancer cell-induced neutrophil activation to increase MMP-9 expression through interacting and stabilizing ALOX5 by post-translational protein modification (PARylation). Blocking PARP-1 by gene knockdown or AG14361 significantly decreased ALOX5 expression and MMP-9 production, and eliminated neutrophil-mediated lung cancer cell invasion and in vivo tumor growth.
We identified a novel mechanism by which PARP-1 mediates lung cancer cell-induced neutrophil activation and PARylates ALOX5 to regulate MMP-9 expression, which exacerbates lung cancer progression.
摘要：
目的：中性粒细胞是肺癌组织中最主要的浸润白细胞之一，与肺癌的进展有关。中性粒细胞如何促进肺癌进展，然而,尚未建立。
方法：使用Kaplan-Meier绘图仪在线分析和组织免疫组织化学来确定中性粒细胞与肺癌患者总生存期之间的关系。嗜中性粒细胞对肺癌的影响是使用Transwell迁移测定法确定的，增殖试验，和鼠肿瘤模型.基因敲低用于确定肺癌-嗜中性粒细胞中聚ADP-核糖聚合酶（PARP）-1的功能。使用蛋白质印迹分析和明胶酶谱来证明PARP-1和基质金属肽酶9(MMP-9)之间的相关性。免疫沉淀与质谱联用(IP/MS)用于鉴定与PARP-1相互作用的蛋白质。免疫共沉淀(Co-IP)用于确认PARP-1与花生四烯酸酯5-脂加氧酶(ALOX5)相互作用。AG14361阻断中性粒细胞PARP-1可挽救中性粒细胞促进肺癌进展。
结果：在肺癌患者中，浸润中性粒细胞数量的增加与总生存期呈负相关（P<0.001）。中性粒细胞活化促进肺癌细胞侵袭,迁移,和体外增殖，和小鼠肺癌在体内的生长。机械上,PARP-1被证明参与肺癌细胞诱导的中性粒细胞活化，通过翻译后蛋白质修饰（PARylation）相互作用和稳定ALOX5来增加MMP-9的表达。通过基因敲低或AG14361阻断PARP-1显著降低ALOX5表达和MMP-9产生,并消除中性粒细胞介导的肺癌细胞侵袭和体内肿瘤生长。
结论：我们确定了PARP-1介导肺癌细胞诱导的中性粒细胞活化和PARylatesALOX5调节MMP-9表达的新机制，这加剧了肺癌的进展。
方法：使用Kaplan-Meier绘图仪在线分析和组织免疫组织化学来确定中性粒细胞与肺癌患者总生存期之间的关系。嗜中性粒细胞对肺癌的影响是使用Transwell迁移测定法确定的，增殖试验，和鼠肿瘤模型.基因敲低用于确定肺癌-嗜中性粒细胞中聚ADP-核糖聚合酶（PARP）-1的功能。使用蛋白质印迹分析和明胶酶谱来证明PARP-1和基质金属肽酶9(MMP-9)之间的相关性。免疫沉淀与质谱联用(IP/MS)用于鉴定与PARP-1相互作用的蛋白质。免疫共沉淀(Co-IP)用于确认PARP-1与花生四烯酸酯5-脂加氧酶(ALOX5)相互作用。AG14361阻断中性粒细胞PARP-1可挽救中性粒细胞促进肺癌进展。
结果：在肺癌患者中，浸润中性粒细胞数量的增加与总生存期呈负相关（P<0.001）。中性粒细胞活化促进肺癌细胞侵袭,迁移,和体外增殖，和小鼠肺癌在体内的生长。机械上,PARP-1被证明参与肺癌细胞诱导的中性粒细胞活化，通过翻译后蛋白质修饰（PARylation）相互作用和稳定ALOX5来增加MMP-9的表达。通过基因敲低或AG14361阻断PARP-1显著降低ALOX5表达和MMP-9产生,并消除中性粒细胞介导的肺癌细胞侵袭和体内肿瘤生长。
结论：我们确定了PARP-1介导肺癌细胞诱导的中性粒细胞活化和PARylatesALOX5调节MMP-9表达的新机制，这加剧了肺癌的进展。
