Colitis-associated colorectal cancer(CAC)is a major contributor to cancer-related mortality worldwide.Titanium dioxide(TiO_(2),E171),a widely used food additive,has been insufficiently studied regarding its effects on...Colitis-associated colorectal cancer(CAC)is a major contributor to cancer-related mortality worldwide.Titanium dioxide(TiO_(2),E171),a widely used food additive,has been insufficiently studied regarding its effects on macrophages within colon tumors during CAC development.In this study,CAC mouse models were used to investigate the biological impact of dietary E171 on macrophages in vivo,while lipopolysaccharide(LPS)-stimulated RAW264.7 macrophage cell lines were employed to elucidate the underlying mechanisms in vitro.We found that dietary E171 intake accelerated CAC development,exacerbated inflammatory responses and oxidative stress,and upregulated CAC-associated genes,including S100a8,S100a9,Lcn2,S100a11,Cxcl2,and interleukin-1α(Il-1α).E171 also increased the expression of S100A8,S100A9,NOD-like receptor family pyrin domain-containing 3(NLRP3),and gasdermin-D Nterminal(GSDMD-N)in macrophages within colon tumors.In inflammatory macrophages,E171 exposure enhanced cell viability,increased reactive oxygen species(ROS)levels,and elevated the expression and secretion of S100A8 and S100A9,consistent with in vivo histological observations.Furthermore,E171-induced secretion of S100A8 and S100A9 in macrophages was suppressed by specific inhibitors,including N-acetylcysteine(NAC,ROS inhibitor),MCC950(NLRP3 inhibitor),Z-YVAD-FMK(caspase 1 inhibitor),disulfiram(GSDMD inhibitor),and transfection of NLRP3 small interfering ribonucleic acid(siRNA).These results indicate that dietary E171 promotes CAC development by activating macrophages,with S100A8 and S100A9 serving as key mediators,and the NLRP3/caspase 1/GSDMD pathway acting as a critical mechanism.展开更多
Objective To investigate the effects of calprotectin(S100A8/A9)on the biological activity of acute myeloid leukemia(AML)cells harboring a DNA methyltransferase 3A(DNMT3A)mutation and to explore the underlying molecula...Objective To investigate the effects of calprotectin(S100A8/A9)on the biological activity of acute myeloid leukemia(AML)cells harboring a DNA methyltransferase 3A(DNMT3A)mutation and to explore the underlying molecular mechanisms involved.Methods AML monoclonal cell lines harboring the DNMT3A^(R882H) mutation were generated via lentiviral transduction and limiting dilution.RNA sequencing was used for differential gene expression analysis,followed by bioinformatic pathway enrichment and gene correlation analyses.The biological effects of paquinimod,a selective S100A8/A9 inhibitor,on DNMT3A^(R882H) AML cells were assessed via Cell Counting Kit(CCK-8)proliferation assays,Annexin V/PI staining,cell cycle analysis,cell adhesion assays,and transwell migration assays.Results Differential gene expression analysis revealed 442 upregulated and 535 downregulated genes in DNMT3A-mutated(DNMT3A^(mut))cells compared with those in DNMT3A wild-type(DNMT3A^(wt))cells,with the S100A8/A9 complex recurrently enriched in Reactome pathway analysis.Compared with healthy controls,patients with AML presented increased expression of S100A8 and S100A9 and increased expression of DNMT3A^(mut) cells relative to DNMT3A^(wt) cells,which was correlated with poor prognosis in patients with AML.There were no notable differences in proliferation among the DNMT3A^(mut),DNMT3A^(wt),and empty vector cells under normal or starvation conditions.However,paquinimod treatment notably inhibited the proliferation,migration,and adhesion of DNMT3A^(mut) AML cells in a dose-dependent manner,causing G0/G1 cell cycle arrest,whereas no significant effects on apoptosis were observed.Paquinimod also downregulated key adhesion molecules,including intercellular adhesion molecule 1(ICAM-1),vascular cell adhesion molecule 1(VCAM-1),monocyte chemoattractant protein-1(MCP-1),and matrix metalloproteinase-2(MMP-2).Additionally,S100A8 and S100A9 expression was upregulated in a dose-dependent manner in response to cytarabine treatment.Conclusion Elevated S100A8/A9 expression contributes to the abnormal proliferation,migration,adhesion,and chemoresistance of DNMT3A^(mut) AML cells.Targeting S100A8/A9 alone or in combination with other treatments represents a promising therapeutic strategy for DNMT3A^(mut) AML.展开更多
基金supported by the National Natural Science Foundation of China(Nos.81974441 and 82203619)the Science and Technology Planning Project of Shenzhen Municipality(Nos.JCYJ20190814105619048 and JCYJ20220530154202005)。
文摘Colitis-associated colorectal cancer(CAC)is a major contributor to cancer-related mortality worldwide.Titanium dioxide(TiO_(2),E171),a widely used food additive,has been insufficiently studied regarding its effects on macrophages within colon tumors during CAC development.In this study,CAC mouse models were used to investigate the biological impact of dietary E171 on macrophages in vivo,while lipopolysaccharide(LPS)-stimulated RAW264.7 macrophage cell lines were employed to elucidate the underlying mechanisms in vitro.We found that dietary E171 intake accelerated CAC development,exacerbated inflammatory responses and oxidative stress,and upregulated CAC-associated genes,including S100a8,S100a9,Lcn2,S100a11,Cxcl2,and interleukin-1α(Il-1α).E171 also increased the expression of S100A8,S100A9,NOD-like receptor family pyrin domain-containing 3(NLRP3),and gasdermin-D Nterminal(GSDMD-N)in macrophages within colon tumors.In inflammatory macrophages,E171 exposure enhanced cell viability,increased reactive oxygen species(ROS)levels,and elevated the expression and secretion of S100A8 and S100A9,consistent with in vivo histological observations.Furthermore,E171-induced secretion of S100A8 and S100A9 in macrophages was suppressed by specific inhibitors,including N-acetylcysteine(NAC,ROS inhibitor),MCC950(NLRP3 inhibitor),Z-YVAD-FMK(caspase 1 inhibitor),disulfiram(GSDMD inhibitor),and transfection of NLRP3 small interfering ribonucleic acid(siRNA).These results indicate that dietary E171 promotes CAC development by activating macrophages,with S100A8 and S100A9 serving as key mediators,and the NLRP3/caspase 1/GSDMD pathway acting as a critical mechanism.
基金funded by the National Natural Science Foundation of China(Grant No.82270177)the China Medicine Education Association 2024 Medical Science and Technology Research Project(Grant No.2024KTZ035).
文摘Objective To investigate the effects of calprotectin(S100A8/A9)on the biological activity of acute myeloid leukemia(AML)cells harboring a DNA methyltransferase 3A(DNMT3A)mutation and to explore the underlying molecular mechanisms involved.Methods AML monoclonal cell lines harboring the DNMT3A^(R882H) mutation were generated via lentiviral transduction and limiting dilution.RNA sequencing was used for differential gene expression analysis,followed by bioinformatic pathway enrichment and gene correlation analyses.The biological effects of paquinimod,a selective S100A8/A9 inhibitor,on DNMT3A^(R882H) AML cells were assessed via Cell Counting Kit(CCK-8)proliferation assays,Annexin V/PI staining,cell cycle analysis,cell adhesion assays,and transwell migration assays.Results Differential gene expression analysis revealed 442 upregulated and 535 downregulated genes in DNMT3A-mutated(DNMT3A^(mut))cells compared with those in DNMT3A wild-type(DNMT3A^(wt))cells,with the S100A8/A9 complex recurrently enriched in Reactome pathway analysis.Compared with healthy controls,patients with AML presented increased expression of S100A8 and S100A9 and increased expression of DNMT3A^(mut) cells relative to DNMT3A^(wt) cells,which was correlated with poor prognosis in patients with AML.There were no notable differences in proliferation among the DNMT3A^(mut),DNMT3A^(wt),and empty vector cells under normal or starvation conditions.However,paquinimod treatment notably inhibited the proliferation,migration,and adhesion of DNMT3A^(mut) AML cells in a dose-dependent manner,causing G0/G1 cell cycle arrest,whereas no significant effects on apoptosis were observed.Paquinimod also downregulated key adhesion molecules,including intercellular adhesion molecule 1(ICAM-1),vascular cell adhesion molecule 1(VCAM-1),monocyte chemoattractant protein-1(MCP-1),and matrix metalloproteinase-2(MMP-2).Additionally,S100A8 and S100A9 expression was upregulated in a dose-dependent manner in response to cytarabine treatment.Conclusion Elevated S100A8/A9 expression contributes to the abnormal proliferation,migration,adhesion,and chemoresistance of DNMT3A^(mut) AML cells.Targeting S100A8/A9 alone or in combination with other treatments represents a promising therapeutic strategy for DNMT3A^(mut) AML.
