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
Objective:To investigate expressions of hypoxia-inducible factors-la(HIF-la),transforming growth factor-β1 (TGF-pl),mucinl(Mucl) and matrix metalloproteinase-9(MMP-9) in the placenta collected from the preeclampsia p...Objective:To investigate expressions of hypoxia-inducible factors-la(HIF-la),transforming growth factor-β1 (TGF-pl),mucinl(Mucl) and matrix metalloproteinase-9(MMP-9) in the placenta collected from the preeclampsia patients and normal pregnant women,so as to explore the possible pathophysiological mechanism of preeclampsia. Methods:The placenta villus tissues were obtained from 35 preeclampsia women,including 16 mild preeclampsia and 19 severe preeclampsia,and 20 normal pregnant women,within 5 minutes after placental expulsion.Expressions of HIF-1α,TGF-β1,Mucl and MMP-9 were detected by Western blot.Cellular location was observed by immunohistochemistry. Results:(1) HIF-1αwas mainly located in cytoplasm and nucleus of placental villous syncytiotrophoblast. Expression level of HIF-la in the severe preeclampsia group was significantly higher than that in the mild group or control group(P<0.01).(2) TGF-pl was located in the trophoblast cell and exuviates membrane.Expression level of TGF-pl in the severe and mild preeclampsia groups was significantly higher than that in control group (P<0.05).(3) Mucl was located in trophoblast cell and exuviates membrane.Expression level of MUC1 in the severe preeclampsia group was significantly higher than that in the mild preeclampsia group and control group(P< 0.01).(4) MMP-9 was located in the trophoblast cell and villous stroma,exuviates membrane.Expression levels of MMP-9 in the two preeclampsia groups were lower than that in control group(P>0.05). Conclusion:Expressions of HIF-lα,TGF-β1 and Mucl increased in the placenta of preeclampsia group,while MMP-9 decreased.Mucl can be induced and regulated by HIF-la and TGF-β1.Over-expression of Mucl in placenta can significantly suppress the activation of MMP-9,which may influence the infiltration of trophoblast cells.The increased expression of HIF-lαand TGF-β1 in placenta may induce higher level of Mucl.This study helped us to understand the pathophysiological mechanism of preeclampsia.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.
基金supported by projects from National Foundation of Science(81170559)China 973 Program(2012CB944703)
文摘Objective:To investigate expressions of hypoxia-inducible factors-la(HIF-la),transforming growth factor-β1 (TGF-pl),mucinl(Mucl) and matrix metalloproteinase-9(MMP-9) in the placenta collected from the preeclampsia patients and normal pregnant women,so as to explore the possible pathophysiological mechanism of preeclampsia. Methods:The placenta villus tissues were obtained from 35 preeclampsia women,including 16 mild preeclampsia and 19 severe preeclampsia,and 20 normal pregnant women,within 5 minutes after placental expulsion.Expressions of HIF-1α,TGF-β1,Mucl and MMP-9 were detected by Western blot.Cellular location was observed by immunohistochemistry. Results:(1) HIF-1αwas mainly located in cytoplasm and nucleus of placental villous syncytiotrophoblast. Expression level of HIF-la in the severe preeclampsia group was significantly higher than that in the mild group or control group(P<0.01).(2) TGF-pl was located in the trophoblast cell and exuviates membrane.Expression level of TGF-pl in the severe and mild preeclampsia groups was significantly higher than that in control group (P<0.05).(3) Mucl was located in trophoblast cell and exuviates membrane.Expression level of MUC1 in the severe preeclampsia group was significantly higher than that in the mild preeclampsia group and control group(P< 0.01).(4) MMP-9 was located in the trophoblast cell and villous stroma,exuviates membrane.Expression levels of MMP-9 in the two preeclampsia groups were lower than that in control group(P>0.05). Conclusion:Expressions of HIF-lα,TGF-β1 and Mucl increased in the placenta of preeclampsia group,while MMP-9 decreased.Mucl can be induced and regulated by HIF-la and TGF-β1.Over-expression of Mucl in placenta can significantly suppress the activation of MMP-9,which may influence the infiltration of trophoblast cells.The increased expression of HIF-lαand TGF-β1 in placenta may induce higher level of Mucl.This study helped us to understand the pathophysiological mechanism of preeclampsia.