目的:探讨次乌头碱(HA)通过环磷酸鸟苷-腺苷酸合成酶(cGAS)/干扰素基因刺激因子(STING)信号通路对胃癌细胞增殖、迁移、侵袭和免疫逃逸的影响。方法:将BGC-823细胞分为对照组(NC组)、HA低剂量组(HA-L组,2μmol/L)、HA中剂量组(HA-M组,4...目的:探讨次乌头碱(HA)通过环磷酸鸟苷-腺苷酸合成酶(cGAS)/干扰素基因刺激因子(STING)信号通路对胃癌细胞增殖、迁移、侵袭和免疫逃逸的影响。方法:将BGC-823细胞分为对照组(NC组)、HA低剂量组(HA-L组,2μmol/L)、HA中剂量组(HA-M组,4μmol/L)、HA高剂量组(HA-H组,8μmol/L)、RU.521(cGAS抑制剂)组(1μmol/L)、HA-H+RU.521组(8μmol/L+1μmol/L),CCK-8、克隆形成实验检测细胞增殖;划痕愈合实验检测细胞迁移;Transwell实验检测细胞侵袭;ELISA检测细胞上清中趋化因子配体(CXCL)2、CXCL8水平;Western blot检测细胞中增殖细胞核抗原(PCNA)、基质金属蛋白酶(MMP)-9、cGAS、STING蛋白表达。将上述各组细胞分别与NK细胞共培养24 h,命名为NC共培养组、HA-L共培养组、HA-M共培养组、HA-H共培养组、RU.521共培养组、HA-H+RU.521共培养组,检测NK细胞杀伤力。结果:与NC组比较,HA-L组、HA-M组、HA-H组BGC-823细胞OD450(24 h)(0.87±0.08 vs 0.75±0.06、0.62±0.06、0.41±0.03)、克隆形成率[(47.75±2.13)%vs(41.12±1.81)%、(33.58±1.61)%、(19.95±0.84)%]、划痕愈合率[(43.37±2.08)%vs(36.65±1.54)%、(27.74±1.03)%、(13.36±0.62)%]、侵袭细胞数(78.85±3.67 vs 65.59±2.49、51.52±2.01、22.23±1.36)、CXCL2、CXCL8水平、PCNA、MMP-9蛋白表达降低,cGAS、STING蛋白表达升高(P<0.05);与NC组比较,RU.521组对应指标变化趋势与上述相反(P<0.05);与NC共培养组比较,HA-L共培养组、HA-M共培养组、HA-H共培养组NK细胞杀伤力增强,且呈剂量依赖性(P<0.05);与NC共培养组比较,RU.521共培养组对应指标变化趋势与上述相反(P<0.05);RU.521减弱了高剂量HA对BGC-823细胞增殖、迁移、侵袭、免疫逃逸的抑制作用。结论:HA可能通过激活cGAS-STING信号通路抑制胃癌细胞增殖、迁移、侵袭和免疫逃逸。展开更多
Cyclic GMP-AMP(cGAMP)synthase(cGAS)plays a pivotal role in the innate immune system.As the primary DNA sensor in cells,cGAS binds to dsDNA in the cytoplasm and forms cGAS-DNA liquid-liquid phase separation(LLPS)and ac...Cyclic GMP-AMP(cGAMP)synthase(cGAS)plays a pivotal role in the innate immune system.As the primary DNA sensor in cells,cGAS binds to dsDNA in the cytoplasm and forms cGAS-DNA liquid-liquid phase separation(LLPS)and activates its catalytic activity.This activation triggers the cGAS-stimulator of interferon genes(STING)signaling pathway,establishing an efficient system for pathogen detection.Beyond pathogen surveillance,cGAS performs a diverse range of roles,involved in inflammatory response,metabolic homeostasis,DNA damage repair,and cell death.These biological functions regulate cellular physiological homeostasis and influence the occurrence and development of diseases.This review provides an overview of the structure,localization,and intracellular biological functions of the cGAS-STING signaling pathway and cGAS-DNA LLPS.Furthermore,we discuss their contribution to the development of tumors,autoimmune diseases,and inflammatory diseases and highlight the innovative strategies in modulating cGAS activity,either through activation or inhibition,as a promising therapeutic approach.展开更多
Pattern recognition receptors arecritical forthe sensing of pathogen-associated molecular patterns or danger-associated molecular patterns and subsequent mounting of innate immunityandshaping ofadaptive immunity.The i...Pattern recognition receptors arecritical forthe sensing of pathogen-associated molecular patterns or danger-associated molecular patterns and subsequent mounting of innate immunityandshaping ofadaptive immunity.The identification of 2'3'-cyclic guanosine monophosphate-adenosine monophosphate(cGAMP)synthase(cGAS)as a major cytosolic DNA receptor is a milestone in the field of DNA sensing.The engagement of cGAS by double-stranded DNA from different origins,including invading pathogens,damaged mitochondria,ruptured micronuclei,and genomic DNA results in the generation of cGAMP and activation of stimulator of interferon genes,which thereby activates innate immunity mainly characterized by the activation of type I interferon response.In recent years,great progress has been made in understanding the subcellular localization and novel functions of cGAS.In this review,we particularlyfocus on summarizingthe multifaceted roles ofcGAS in regulating senescence,autophagy,cell stemness,apoptosis,angiogenesis,cell proliferation,antitumor effect,DNA replication,DNA damage repair,micronucleophagy,as well as cell metabolism.展开更多
文摘目的:探讨次乌头碱(HA)通过环磷酸鸟苷-腺苷酸合成酶(cGAS)/干扰素基因刺激因子(STING)信号通路对胃癌细胞增殖、迁移、侵袭和免疫逃逸的影响。方法:将BGC-823细胞分为对照组(NC组)、HA低剂量组(HA-L组,2μmol/L)、HA中剂量组(HA-M组,4μmol/L)、HA高剂量组(HA-H组,8μmol/L)、RU.521(cGAS抑制剂)组(1μmol/L)、HA-H+RU.521组(8μmol/L+1μmol/L),CCK-8、克隆形成实验检测细胞增殖;划痕愈合实验检测细胞迁移;Transwell实验检测细胞侵袭;ELISA检测细胞上清中趋化因子配体(CXCL)2、CXCL8水平;Western blot检测细胞中增殖细胞核抗原(PCNA)、基质金属蛋白酶(MMP)-9、cGAS、STING蛋白表达。将上述各组细胞分别与NK细胞共培养24 h,命名为NC共培养组、HA-L共培养组、HA-M共培养组、HA-H共培养组、RU.521共培养组、HA-H+RU.521共培养组,检测NK细胞杀伤力。结果:与NC组比较,HA-L组、HA-M组、HA-H组BGC-823细胞OD450(24 h)(0.87±0.08 vs 0.75±0.06、0.62±0.06、0.41±0.03)、克隆形成率[(47.75±2.13)%vs(41.12±1.81)%、(33.58±1.61)%、(19.95±0.84)%]、划痕愈合率[(43.37±2.08)%vs(36.65±1.54)%、(27.74±1.03)%、(13.36±0.62)%]、侵袭细胞数(78.85±3.67 vs 65.59±2.49、51.52±2.01、22.23±1.36)、CXCL2、CXCL8水平、PCNA、MMP-9蛋白表达降低,cGAS、STING蛋白表达升高(P<0.05);与NC组比较,RU.521组对应指标变化趋势与上述相反(P<0.05);与NC共培养组比较,HA-L共培养组、HA-M共培养组、HA-H共培养组NK细胞杀伤力增强,且呈剂量依赖性(P<0.05);与NC共培养组比较,RU.521共培养组对应指标变化趋势与上述相反(P<0.05);RU.521减弱了高剂量HA对BGC-823细胞增殖、迁移、侵袭、免疫逃逸的抑制作用。结论:HA可能通过激活cGAS-STING信号通路抑制胃癌细胞增殖、迁移、侵袭和免疫逃逸。
基金supported by the National Natural Science Foundation of China(Nos.22274166,22021003,22234008,21927807)the Project of the Youth Innovation Promotion Association,Chinese Academy of Sciences(No.2023047).
文摘Cyclic GMP-AMP(cGAMP)synthase(cGAS)plays a pivotal role in the innate immune system.As the primary DNA sensor in cells,cGAS binds to dsDNA in the cytoplasm and forms cGAS-DNA liquid-liquid phase separation(LLPS)and activates its catalytic activity.This activation triggers the cGAS-stimulator of interferon genes(STING)signaling pathway,establishing an efficient system for pathogen detection.Beyond pathogen surveillance,cGAS performs a diverse range of roles,involved in inflammatory response,metabolic homeostasis,DNA damage repair,and cell death.These biological functions regulate cellular physiological homeostasis and influence the occurrence and development of diseases.This review provides an overview of the structure,localization,and intracellular biological functions of the cGAS-STING signaling pathway and cGAS-DNA LLPS.Furthermore,we discuss their contribution to the development of tumors,autoimmune diseases,and inflammatory diseases and highlight the innovative strategies in modulating cGAS activity,either through activation or inhibition,as a promising therapeutic approach.
基金This work was supported by the National Natural Science Foundation of China(81922030 and 81770006 to H.L,32188101,32030038,91842303,and 31730025 to B.G.)the Major Research Plan of National Natural Science Foundation of China(2017YFA0505900 to B.G.),Shanghai Shu Guang Program(205G19)Shanghai Science and Technology Fund(19140900600 and 22S11900700 to H.L.).
文摘Pattern recognition receptors arecritical forthe sensing of pathogen-associated molecular patterns or danger-associated molecular patterns and subsequent mounting of innate immunityandshaping ofadaptive immunity.The identification of 2'3'-cyclic guanosine monophosphate-adenosine monophosphate(cGAMP)synthase(cGAS)as a major cytosolic DNA receptor is a milestone in the field of DNA sensing.The engagement of cGAS by double-stranded DNA from different origins,including invading pathogens,damaged mitochondria,ruptured micronuclei,and genomic DNA results in the generation of cGAMP and activation of stimulator of interferon genes,which thereby activates innate immunity mainly characterized by the activation of type I interferon response.In recent years,great progress has been made in understanding the subcellular localization and novel functions of cGAS.In this review,we particularlyfocus on summarizingthe multifaceted roles ofcGAS in regulating senescence,autophagy,cell stemness,apoptosis,angiogenesis,cell proliferation,antitumor effect,DNA replication,DNA damage repair,micronucleophagy,as well as cell metabolism.
