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
钙结合蛋白S100A(calcium binding protein S100A,S100A)4是经RNA互作组技术发现的一种新型RNA结合蛋白(RNA-binding proteins,RBPs)[1],能够识别特定RNA结合域并与RNA相互作用,在转录后基因表达调控中起着重要作用[2]。先前的研究发现,...钙结合蛋白S100A(calcium binding protein S100A,S100A)4是经RNA互作组技术发现的一种新型RNA结合蛋白(RNA-binding proteins,RBPs)[1],能够识别特定RNA结合域并与RNA相互作用,在转录后基因表达调控中起着重要作用[2]。先前的研究发现,S100A4不仅在银屑病患者皮肤的真皮组织中表达上调,而且能通过影响表皮细胞增殖和真皮血管增生参与银屑病的发生发展,然而其中潜在的调控机制尚不清楚[3]。展开更多
[目的]探究S100A12、RAGE在宫颈癌细胞增殖、迁移和凋亡中的作用。[方法]采集2022年4月-2023年5月宫颈癌病例癌组织及癌旁组织的石蜡标本,用免疫组化法检测S100A12的表达。随机将SiHa细胞分为空白组(无转染)、阴性对照组(转染空白质粒)...[目的]探究S100A12、RAGE在宫颈癌细胞增殖、迁移和凋亡中的作用。[方法]采集2022年4月-2023年5月宫颈癌病例癌组织及癌旁组织的石蜡标本,用免疫组化法检测S100A12的表达。随机将SiHa细胞分为空白组(无转染)、阴性对照组(转染空白质粒)和敲低组(转染siRNA),构建相应细胞模型。免疫组化检测S100A12在宫颈癌组织和癌旁组织的表达情况;实时荧光定量PCR检测S100A12 mRNA与RAGE mRNA水平;Western Blot检测PCNA、S100A12的蛋白表达水平;通过MTT法以及细胞集落形成实验检测细胞的生长情况,通过Transwell实验检测细胞迁移情况;流式细胞术检测细胞凋亡情况。[结果]S100A12在宫颈癌组织中蛋白表达水平高于癌旁组织(82.47%±0.35%vs 11.34%±0.17%,P<0.05)。敲低组细胞中S100A12、RAGE的mRNA和蛋白表达均低于空白组与阴性对照组(1.16±0.12 vs 1.13±0.16 vs 0.17±0.08;2.57±0.13 vs 2.67±0.13 vs 0.65±0.05;P<0.05)。敲低组细胞的增殖能力、集落形成数量和迁移能力均低于空白组、阴性对照组(P<0.05)。敲低组细胞的凋亡率高于空白组、阴性对照组(1.16%±0.09%vs 1.78%±0.03%vs 11.53%±0.09%;P<0.05)。[结论]宫颈癌组织中RAGE与S100A12表达量显著升高,下调S100A12后,癌细胞的增殖、迁移能力减弱,凋亡率增加,并且RAGE表达受到抑制。因此,S100A12对宫颈癌细胞的抑制作用与调控RAGE相关。展开更多
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.
文摘钙结合蛋白S100A(calcium binding protein S100A,S100A)4是经RNA互作组技术发现的一种新型RNA结合蛋白(RNA-binding proteins,RBPs)[1],能够识别特定RNA结合域并与RNA相互作用,在转录后基因表达调控中起着重要作用[2]。先前的研究发现,S100A4不仅在银屑病患者皮肤的真皮组织中表达上调,而且能通过影响表皮细胞增殖和真皮血管增生参与银屑病的发生发展,然而其中潜在的调控机制尚不清楚[3]。
文摘[目的]探究S100A12、RAGE在宫颈癌细胞增殖、迁移和凋亡中的作用。[方法]采集2022年4月-2023年5月宫颈癌病例癌组织及癌旁组织的石蜡标本,用免疫组化法检测S100A12的表达。随机将SiHa细胞分为空白组(无转染)、阴性对照组(转染空白质粒)和敲低组(转染siRNA),构建相应细胞模型。免疫组化检测S100A12在宫颈癌组织和癌旁组织的表达情况;实时荧光定量PCR检测S100A12 mRNA与RAGE mRNA水平;Western Blot检测PCNA、S100A12的蛋白表达水平;通过MTT法以及细胞集落形成实验检测细胞的生长情况,通过Transwell实验检测细胞迁移情况;流式细胞术检测细胞凋亡情况。[结果]S100A12在宫颈癌组织中蛋白表达水平高于癌旁组织(82.47%±0.35%vs 11.34%±0.17%,P<0.05)。敲低组细胞中S100A12、RAGE的mRNA和蛋白表达均低于空白组与阴性对照组(1.16±0.12 vs 1.13±0.16 vs 0.17±0.08;2.57±0.13 vs 2.67±0.13 vs 0.65±0.05;P<0.05)。敲低组细胞的增殖能力、集落形成数量和迁移能力均低于空白组、阴性对照组(P<0.05)。敲低组细胞的凋亡率高于空白组、阴性对照组(1.16%±0.09%vs 1.78%±0.03%vs 11.53%±0.09%;P<0.05)。[结论]宫颈癌组织中RAGE与S100A12表达量显著升高,下调S100A12后,癌细胞的增殖、迁移能力减弱,凋亡率增加,并且RAGE表达受到抑制。因此,S100A12对宫颈癌细胞的抑制作用与调控RAGE相关。
