Objective: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...Objective: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.Methods: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 ADPribose 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.Results: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.Conclusion: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.展开更多
目的探讨花生四烯5-脂氧合酶基因(arachidonate 5-lipoxygenase gene, ALOX5)基因启动子SP-1结合位点的多态性与成人急性髓系白血病(acute myeloid leukemia, AML)的相关性。方法 采用聚合酶链反应-单链构象多态性(PCR-SSCP)分析236例成...目的探讨花生四烯5-脂氧合酶基因(arachidonate 5-lipoxygenase gene, ALOX5)基因启动子SP-1结合位点的多态性与成人急性髓系白血病(acute myeloid leukemia, AML)的相关性。方法 采用聚合酶链反应-单链构象多态性(PCR-SSCP)分析236例成人AML(实验组)和179例健康体检者(对照组)中ALOX5基因启动子SP-1结合位点的多态性,经PCR产物直接测序法进行验证,利用DNAStar软件对测序结果进行分析。结果 检测到ALOX5基因启动子SP-1结合位点有6种基因型,包括4/4、4/5、4/6、5/5、5/6和6/6。在实验组和对照组中,基因型4/4的分布频率为20.30%和10.10%( OR =1.976,95% CI :1.050~3.719,χ 2=4.539, P <0.05);基因型4/6的分布频率为10.60%和26.80%( OR =0.386,95% CI :0.215~0.692,χ 2=10.513, P <0.05);等位基因6的分布频率为26.90%和30.20%( OR =0.736,95% CI :0.528~1.025,χ 2=3.291, P <0.05);SP-1位点的其余基因型、等位基因在实验组和对照组中分布频率差异无统计学意义( P >0.05),其可能不是成人AML发病风险的预警指标。结论 ALOX5基因启动子区SP-1结合位点的基因型4/6,等位基因6可能与AML发病风险降低有关,而携带基因型4/4人群可能易患AML。展开更多
基金supported by grants from the National Key R&D Program of China(Grant No.2018YFA0900900)the National Natural Science Foundation of China(Grant Nos.82273334,82203172,81871869,and 81400055)+3 种基金the Jiangsu Province Social Development Key Projects(Grant Nos.BE2020641 and BE2020640)the Xuzhou Medical University Excellent Talent Research Start-up Fund(Grant No.RC20552157)the Jiangsu Province Capability Improvement Project through Science,Technology and Education(Grant No.CXZX202234)funded by the China Postdoctoral Science Foundation(Grant No.2023M732970)。
文摘Objective: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.Methods: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 ADPribose 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.Results: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.Conclusion: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.
文摘目的探讨花生四烯5-脂氧合酶基因(arachidonate 5-lipoxygenase gene, ALOX5)基因启动子SP-1结合位点的多态性与成人急性髓系白血病(acute myeloid leukemia, AML)的相关性。方法 采用聚合酶链反应-单链构象多态性(PCR-SSCP)分析236例成人AML(实验组)和179例健康体检者(对照组)中ALOX5基因启动子SP-1结合位点的多态性,经PCR产物直接测序法进行验证,利用DNAStar软件对测序结果进行分析。结果 检测到ALOX5基因启动子SP-1结合位点有6种基因型,包括4/4、4/5、4/6、5/5、5/6和6/6。在实验组和对照组中,基因型4/4的分布频率为20.30%和10.10%( OR =1.976,95% CI :1.050~3.719,χ 2=4.539, P <0.05);基因型4/6的分布频率为10.60%和26.80%( OR =0.386,95% CI :0.215~0.692,χ 2=10.513, P <0.05);等位基因6的分布频率为26.90%和30.20%( OR =0.736,95% CI :0.528~1.025,χ 2=3.291, P <0.05);SP-1位点的其余基因型、等位基因在实验组和对照组中分布频率差异无统计学意义( P >0.05),其可能不是成人AML发病风险的预警指标。结论 ALOX5基因启动子区SP-1结合位点的基因型4/6,等位基因6可能与AML发病风险降低有关,而携带基因型4/4人群可能易患AML。
