Our previous study has demonstrated that lnc_000048 is upregulated in large-artery atherosclerotic stroke and promotes atherosclerosis in ApoE^(-/-)mice.However,little is known about the role of lnc_000048 in classica...Our previous study has demonstrated that lnc_000048 is upregulated in large-artery atherosclerotic stroke and promotes atherosclerosis in ApoE^(-/-)mice.However,little is known about the role of lnc_000048 in classically activated macrophage(M1)polarization.In this study,we established THP-1-derived testing state macrophages(M0),M1 macrophages,and alternately activated macrophages(M2).Real-time fluorescence quantitative PCR was used to verify the expression of marker genes and the expression of lnc_000048 in macrophages.Flow cytometry was used to detect phenotypic proteins(CD11b,CD38,CD80).We generated cell lines with lentivirus-mediated upregulation or downregulation of lnc_000048.Flow cytometry,western blot,and real-time fluorescence quantitative PCR results showed that down-regulation of lnc_000048 reduced M1 macrophage polarization and the inflammation response,while over-expression of lnc_000048 led to the opposite effect.Western blot results indicated that lnc_000048 enhanced the activation of the STAT1 pathway and mediated the M1 macrophage polarization.Moreover,catRAPID prediction,RNA-pull down,and mass spectrometry were used to identify and screen the protein kinase RNA-activated(PKR),then catRAPID and RPIseq were used to predict the binding ability of lnc_000048 to PKR.Immunofluorescence(IF)-RNA fluorescence in situ hybridization(FISH)double labeling was performed to verify the subcellular colocalization of lnc_000048 and PKR in the cytoplasm of M1 macrophage.We speculate that lnc_000048 may form stem-loop structure-specific binding and activate PKR by inducing its phosphorylation,leading to activation of STAT1 phosphorylation and thereby enhancing STAT1 pathway-mediated polarization of THP-1 macrophages to M1 and inflammatory factor expression.Taken together,these results reveal that the lnc_000048/PKR/STAT1 axis plays a crucial role in the polarization of M1 macrophages and may be a novel therapeutic target for atherosclerosis alleviation in stroke.展开更多
乳腺癌是全球三大最常见癌症之一,也是女性有关癌症死亡的主要原因。由于公众的密切关注对乳腺癌的识别和筛查产生了积极影响,治疗上也取得了长足进展,避免过度治疗和治疗不足已成为一个焦点,治疗方法需要在多学科背景下确定,同时还要...乳腺癌是全球三大最常见癌症之一,也是女性有关癌症死亡的主要原因。由于公众的密切关注对乳腺癌的识别和筛查产生了积极影响,治疗上也取得了长足进展,避免过度治疗和治疗不足已成为一个焦点,治疗方法需要在多学科背景下确定,同时还要考虑分子亚型和局部肿瘤负荷。近年来,信号传导及转录激活蛋白(signal transduction and activator of transcription,STAT)在肿瘤中的应用受到了广泛关注,它们参与肿瘤的发生发展,包括细胞增殖和凋亡、细胞周期进展、血管生成、肿瘤细胞的转移以及免疫逃逸等过程。STAT1可能与肿瘤的侵袭性生物学行为相关[1],在不同肿瘤或者同种肿瘤的不同阶段具有差异性表达,推测肿瘤细胞会根据特定的遗传背景选择性激活或抑制STAT1的表达[2]。本文对STAT1在乳腺癌中主要的促癌和抑癌功能及其机制进行概述。展开更多
目的:研究白细胞介素27(interleukin-27,IL-27)对辅助性T细胞1(helper T cell 1,Th1)分化及急性肺损伤的影响与作用机制。方法:通过盲肠结扎穿刺(cecal ligation and puncture,CLP)构建脓毒症急性肺损伤小鼠模型,苏木精-伊红(hematoxyli...目的:研究白细胞介素27(interleukin-27,IL-27)对辅助性T细胞1(helper T cell 1,Th1)分化及急性肺损伤的影响与作用机制。方法:通过盲肠结扎穿刺(cecal ligation and puncture,CLP)构建脓毒症急性肺损伤小鼠模型,苏木精-伊红(hematoxylin-eosin staining,HE)染色评估肺组织病理损伤。采用Western Blot、RT-qPCR检测JAK/STAT通路表达与激活以及T盒转录因子21(T-box transcription factor 21,T-bet)蛋白的表达。结果:IL-27加重脓毒症小鼠肺损伤。细胞实验表明IL-27显著激活JAK2/STAT1通路磷酸化,上调T-bet蛋白(P<0.01)和mRNA水平(P<0.05)。抑制STAT1磷酸化后,T-bet表达显著下调(P<0.01)。结论:IL-27通过JAK2/STAT1通路促进Th1分化,加重脓毒症小鼠急性肺损伤。展开更多
基金supported by the Natural Science Foundation of Shandong Province,No.ZR2020MH138(to XZ).
文摘Our previous study has demonstrated that lnc_000048 is upregulated in large-artery atherosclerotic stroke and promotes atherosclerosis in ApoE^(-/-)mice.However,little is known about the role of lnc_000048 in classically activated macrophage(M1)polarization.In this study,we established THP-1-derived testing state macrophages(M0),M1 macrophages,and alternately activated macrophages(M2).Real-time fluorescence quantitative PCR was used to verify the expression of marker genes and the expression of lnc_000048 in macrophages.Flow cytometry was used to detect phenotypic proteins(CD11b,CD38,CD80).We generated cell lines with lentivirus-mediated upregulation or downregulation of lnc_000048.Flow cytometry,western blot,and real-time fluorescence quantitative PCR results showed that down-regulation of lnc_000048 reduced M1 macrophage polarization and the inflammation response,while over-expression of lnc_000048 led to the opposite effect.Western blot results indicated that lnc_000048 enhanced the activation of the STAT1 pathway and mediated the M1 macrophage polarization.Moreover,catRAPID prediction,RNA-pull down,and mass spectrometry were used to identify and screen the protein kinase RNA-activated(PKR),then catRAPID and RPIseq were used to predict the binding ability of lnc_000048 to PKR.Immunofluorescence(IF)-RNA fluorescence in situ hybridization(FISH)double labeling was performed to verify the subcellular colocalization of lnc_000048 and PKR in the cytoplasm of M1 macrophage.We speculate that lnc_000048 may form stem-loop structure-specific binding and activate PKR by inducing its phosphorylation,leading to activation of STAT1 phosphorylation and thereby enhancing STAT1 pathway-mediated polarization of THP-1 macrophages to M1 and inflammatory factor expression.Taken together,these results reveal that the lnc_000048/PKR/STAT1 axis plays a crucial role in the polarization of M1 macrophages and may be a novel therapeutic target for atherosclerosis alleviation in stroke.
文摘乳腺癌是全球三大最常见癌症之一,也是女性有关癌症死亡的主要原因。由于公众的密切关注对乳腺癌的识别和筛查产生了积极影响,治疗上也取得了长足进展,避免过度治疗和治疗不足已成为一个焦点,治疗方法需要在多学科背景下确定,同时还要考虑分子亚型和局部肿瘤负荷。近年来,信号传导及转录激活蛋白(signal transduction and activator of transcription,STAT)在肿瘤中的应用受到了广泛关注,它们参与肿瘤的发生发展,包括细胞增殖和凋亡、细胞周期进展、血管生成、肿瘤细胞的转移以及免疫逃逸等过程。STAT1可能与肿瘤的侵袭性生物学行为相关[1],在不同肿瘤或者同种肿瘤的不同阶段具有差异性表达,推测肿瘤细胞会根据特定的遗传背景选择性激活或抑制STAT1的表达[2]。本文对STAT1在乳腺癌中主要的促癌和抑癌功能及其机制进行概述。
