Acute respiratory distress syndrome(ARDS)is one of the most fatal diseases worldwide.Pulmonary fibrosis occurs early in ARDS,and its severity plays a crucial role in ARDS mortality rate.Some studies suggested that fib...Acute respiratory distress syndrome(ARDS)is one of the most fatal diseases worldwide.Pulmonary fibrosis occurs early in ARDS,and its severity plays a crucial role in ARDS mortality rate.Some studies suggested that fibroproliferation is an essential mechanism in ARDS.Mitofusion2(Mfn2)overexpression plays a role in inhibiting cell proliferation.However,the role and potential mechanism of Mfn2 on the proliferation of fibroblasts is still unknown.In this study,we aimed at exploring the effect of Mfn2 on the human embryonic lung fibroblasts(HELF)and discussed its related mechanism.The HELF were treated with the Mfn2 overexpressing lentivirus(adv-Mfn2).The cell cycle was detected by flow cytometry.MTT,PCR and Western blotting were used to investigate the effect of Mfn2 on the proliferation of the HELF,collagen expression,the RAS-RAF-1-ERK1/2 pathway and the expression of cycle-related proteins(p21,p27,Rb,Raf-1,p-Raf-1,Erk1/2 and p-Erk1/2).The co-immunoprecipitation assay was used to explore the interaction between Mfn2 and Ras.The results showed that the overexpression of Mfn2 inhibited the proliferation of the HELF and induced the cell cycle arrest at the G0/G1 phase.Meanwhile,Mfn2 also inhibited the expression of collagen I,p-Erk and p-Raf-1.In addition,an interaction between Mfn2 and Ras existed in the HELF.This study suggests that the overexpression of Mfn2 can decrease the proliferation of HELF in ARDS,which was associated with the inhibition of the RAS-RAF-1-ERK1/2 pathway.The results may offer a potential therapeutic intervention for patients with ARDS.展开更多
背景:前期研究发现,电针能有效抑制凋亡调控基因p38及Fas m RNA的表达,并降低MAPK信号通路的表达,从而抑制软骨细胞凋亡,同时电针可影响骨性关节炎软骨细胞JAK-STAT信号通路,并有效增加转化生长因子β1含量及促进关节软骨中STAT3,Smad3...背景:前期研究发现,电针能有效抑制凋亡调控基因p38及Fas m RNA的表达,并降低MAPK信号通路的表达,从而抑制软骨细胞凋亡,同时电针可影响骨性关节炎软骨细胞JAK-STAT信号通路,并有效增加转化生长因子β1含量及促进关节软骨中STAT3,Smad3及Lep R m RNA的表达以延缓关节软骨退变。目的:观察电针对骨关节炎大鼠关节软骨超微结构及软骨组织中Ras,Raf,MEK1/2,ERK1/2 m RNA表达的影响。方法:建立膝骨关节炎大鼠模型,造模成功后2周随机分为4组,模型组不进行电针干预,电针15,30 min组取双侧膝关节内、外膝眼,分别经电针治疗15,30 min,PD98059组经静脉注射细胞外信号调节激酶抑制剂PD98059,另设正常饲养的大鼠作正常组对照,共干预3个月。结果与结论:(1)透射电镜检测显示,与模型组比,PD98059抑制剂组、电针15,30 min组软骨细胞形态变化较小,细胞核较大,部分内质网池扩张,线粒体结构仍较清晰;(2)ELISA结果显示,与模型组比,PD98059抑制剂组、电针15,30 min组的肿瘤坏死因子α水平降低(P<0.01);(3)RT-PCR结果显示,与模型组相比,PD98059抑制剂组、电针15,30 min组的Ras,Raf,MEK1/2及ERK1/2 m RNA表达显著降低(P<0.05或P<0.01);(4)结果说明,电针干预能够减轻膝骨关节炎模型大鼠软骨细胞损伤,降低膝关节部滑膜组织中肿瘤坏死因子α水平,下调关节软骨Ras,Raf,MEK1/2及ERK1/2 m RNA表达,从而对骨关节炎关节软骨发挥一定的保护作用。展开更多
目的探讨转移相关蛋白2(MTA2)对前列腺癌预后的预测价值及在癌转移过程中的作用。方法收集空军军医大学第一附属医院2018年1月~2020年6月收治的50例前列腺癌患者的癌组织及配对癌旁组织样本用于免疫组织化学分析。根据MTA2蛋白的表达水...目的探讨转移相关蛋白2(MTA2)对前列腺癌预后的预测价值及在癌转移过程中的作用。方法收集空军军医大学第一附属医院2018年1月~2020年6月收治的50例前列腺癌患者的癌组织及配对癌旁组织样本用于免疫组织化学分析。根据MTA2蛋白的表达水平将样本分为两组,染色评分≥4为高表达组,<4为低表达组。对人前列腺癌细胞系(PC-3)转染shMTA2(MTA2-shRNA-pLKO.1)来沉默MTA2作为shMTA2组,转染Luc-shRNA-pLKO.1的细胞作为阴性对照组(shNC组),未转染的细胞作为空白对照组(control组)。通过MTT法检测细胞活力,Transwells实验进行体外迁移和侵袭分析。通过qRT-PCR或Western Blot检测细胞中MTA2、Eotaxin-1、CCR3、p-ERK1/2、t-ERK1/2和MMP-3的表达。结果前列腺癌组织中MTA2的染色评分显著高于癌旁组织(5.16±0.87 vs 2.34±0.39,t=9.221,P<0.001)。低表达组患者中出现淋巴结转移率为14.29%(2/14),高表达组患者中出现淋巴结转移率为52.78%(19/36),两组比较差异有统计学意义(χ^(2)=6.131,P=0.013)。与低表达组相比,高表达组患者的生存时间更长(χ^(2)=4.756,P=0.029)。与空白对照组相比,shMTA2组的细胞活力降低了56.87%(P<0.001),细胞迁移数量降低了65.80%(P<0.001),细胞侵袭数量降低了56.35%(P<0.001),Eotaxin-1、CCR3、p-ERK1/2和MMP-3的蛋白相对表达量依次降低了76.03%、69.12%、72.32%和54.67%(P<0.001)。结论前列腺癌组织中MTA2的表达水平升高且与患者的预后有关,沉默MTA2可降低前列腺癌细胞的迁移和侵袭能力,并抑制Eotaxin-CCR3-ERK1/2-MMP-3轴。展开更多
目的:探讨通过阻断花生四烯酸(arachidonic acid,AA)代谢途径抑制胰腺癌细胞增殖.方法:将胰腺癌细胞SW1990分为对照组,M K886干预组、塞莱昔布(C e l e c o x i b)干预组,MK886+Celecoxib干预组,用RT-PCR法检测细胞白三烯B4受体1(leukot...目的:探讨通过阻断花生四烯酸(arachidonic acid,AA)代谢途径抑制胰腺癌细胞增殖.方法:将胰腺癌细胞SW1990分为对照组,M K886干预组、塞莱昔布(C e l e c o x i b)干预组,MK886+Celecoxib干预组,用RT-PCR法检测细胞白三烯B4受体1(leukotriene B4receptor 1,BLT1)mRNA,血管内皮生长因子(vascular endothelial growth factor,VEGF)mRNA的表达量变化,用Western blot检测磷酸化-Erk(phosphorylated-extracellular regulated protein,p-Erk)表达量变化.结果:MK886作用下,BLT1 mRNA、VEGF mRNA等表达量均减少(P<0.01),p-Erk表达量明显减少(P<0.05),Celecoxib作用下,VEGF mRNA表达量明显减少(P<0.01),BLT1 mRNA表达与对照组无明显差异,p-Erk表达量与MK886组比较明显增加(P<0.01),MK886+80?mol/L Celecoxib作用下,BLT1 mRNA、VEGF mRNA表达量明显减少(P<0.01),p-Erk表达量与对照组无明显差异.结论:花生四烯酸的两条代谢途径均与胰腺癌的发生及增殖均有密切关系,而抑制5-脂氧合酶(5-lipoxygenase)途径较环氧化酶2(cyclooxygenase 2)途径相比,抑制肿瘤细胞增殖作用更强.展开更多
基金This project was supported by Wuhan Medical Science Foundation of China(No.WX17B07,No.WX19A09,and No.WJ2019H324).
文摘Acute respiratory distress syndrome(ARDS)is one of the most fatal diseases worldwide.Pulmonary fibrosis occurs early in ARDS,and its severity plays a crucial role in ARDS mortality rate.Some studies suggested that fibroproliferation is an essential mechanism in ARDS.Mitofusion2(Mfn2)overexpression plays a role in inhibiting cell proliferation.However,the role and potential mechanism of Mfn2 on the proliferation of fibroblasts is still unknown.In this study,we aimed at exploring the effect of Mfn2 on the human embryonic lung fibroblasts(HELF)and discussed its related mechanism.The HELF were treated with the Mfn2 overexpressing lentivirus(adv-Mfn2).The cell cycle was detected by flow cytometry.MTT,PCR and Western blotting were used to investigate the effect of Mfn2 on the proliferation of the HELF,collagen expression,the RAS-RAF-1-ERK1/2 pathway and the expression of cycle-related proteins(p21,p27,Rb,Raf-1,p-Raf-1,Erk1/2 and p-Erk1/2).The co-immunoprecipitation assay was used to explore the interaction between Mfn2 and Ras.The results showed that the overexpression of Mfn2 inhibited the proliferation of the HELF and induced the cell cycle arrest at the G0/G1 phase.Meanwhile,Mfn2 also inhibited the expression of collagen I,p-Erk and p-Raf-1.In addition,an interaction between Mfn2 and Ras existed in the HELF.This study suggests that the overexpression of Mfn2 can decrease the proliferation of HELF in ARDS,which was associated with the inhibition of the RAS-RAF-1-ERK1/2 pathway.The results may offer a potential therapeutic intervention for patients with ARDS.
文摘背景:前期研究发现,电针能有效抑制凋亡调控基因p38及Fas m RNA的表达,并降低MAPK信号通路的表达,从而抑制软骨细胞凋亡,同时电针可影响骨性关节炎软骨细胞JAK-STAT信号通路,并有效增加转化生长因子β1含量及促进关节软骨中STAT3,Smad3及Lep R m RNA的表达以延缓关节软骨退变。目的:观察电针对骨关节炎大鼠关节软骨超微结构及软骨组织中Ras,Raf,MEK1/2,ERK1/2 m RNA表达的影响。方法:建立膝骨关节炎大鼠模型,造模成功后2周随机分为4组,模型组不进行电针干预,电针15,30 min组取双侧膝关节内、外膝眼,分别经电针治疗15,30 min,PD98059组经静脉注射细胞外信号调节激酶抑制剂PD98059,另设正常饲养的大鼠作正常组对照,共干预3个月。结果与结论:(1)透射电镜检测显示,与模型组比,PD98059抑制剂组、电针15,30 min组软骨细胞形态变化较小,细胞核较大,部分内质网池扩张,线粒体结构仍较清晰;(2)ELISA结果显示,与模型组比,PD98059抑制剂组、电针15,30 min组的肿瘤坏死因子α水平降低(P<0.01);(3)RT-PCR结果显示,与模型组相比,PD98059抑制剂组、电针15,30 min组的Ras,Raf,MEK1/2及ERK1/2 m RNA表达显著降低(P<0.05或P<0.01);(4)结果说明,电针干预能够减轻膝骨关节炎模型大鼠软骨细胞损伤,降低膝关节部滑膜组织中肿瘤坏死因子α水平,下调关节软骨Ras,Raf,MEK1/2及ERK1/2 m RNA表达,从而对骨关节炎关节软骨发挥一定的保护作用。
文摘目的探讨转移相关蛋白2(MTA2)对前列腺癌预后的预测价值及在癌转移过程中的作用。方法收集空军军医大学第一附属医院2018年1月~2020年6月收治的50例前列腺癌患者的癌组织及配对癌旁组织样本用于免疫组织化学分析。根据MTA2蛋白的表达水平将样本分为两组,染色评分≥4为高表达组,<4为低表达组。对人前列腺癌细胞系(PC-3)转染shMTA2(MTA2-shRNA-pLKO.1)来沉默MTA2作为shMTA2组,转染Luc-shRNA-pLKO.1的细胞作为阴性对照组(shNC组),未转染的细胞作为空白对照组(control组)。通过MTT法检测细胞活力,Transwells实验进行体外迁移和侵袭分析。通过qRT-PCR或Western Blot检测细胞中MTA2、Eotaxin-1、CCR3、p-ERK1/2、t-ERK1/2和MMP-3的表达。结果前列腺癌组织中MTA2的染色评分显著高于癌旁组织(5.16±0.87 vs 2.34±0.39,t=9.221,P<0.001)。低表达组患者中出现淋巴结转移率为14.29%(2/14),高表达组患者中出现淋巴结转移率为52.78%(19/36),两组比较差异有统计学意义(χ^(2)=6.131,P=0.013)。与低表达组相比,高表达组患者的生存时间更长(χ^(2)=4.756,P=0.029)。与空白对照组相比,shMTA2组的细胞活力降低了56.87%(P<0.001),细胞迁移数量降低了65.80%(P<0.001),细胞侵袭数量降低了56.35%(P<0.001),Eotaxin-1、CCR3、p-ERK1/2和MMP-3的蛋白相对表达量依次降低了76.03%、69.12%、72.32%和54.67%(P<0.001)。结论前列腺癌组织中MTA2的表达水平升高且与患者的预后有关,沉默MTA2可降低前列腺癌细胞的迁移和侵袭能力,并抑制Eotaxin-CCR3-ERK1/2-MMP-3轴。
文摘目的:探讨通过阻断花生四烯酸(arachidonic acid,AA)代谢途径抑制胰腺癌细胞增殖.方法:将胰腺癌细胞SW1990分为对照组,M K886干预组、塞莱昔布(C e l e c o x i b)干预组,MK886+Celecoxib干预组,用RT-PCR法检测细胞白三烯B4受体1(leukotriene B4receptor 1,BLT1)mRNA,血管内皮生长因子(vascular endothelial growth factor,VEGF)mRNA的表达量变化,用Western blot检测磷酸化-Erk(phosphorylated-extracellular regulated protein,p-Erk)表达量变化.结果:MK886作用下,BLT1 mRNA、VEGF mRNA等表达量均减少(P<0.01),p-Erk表达量明显减少(P<0.05),Celecoxib作用下,VEGF mRNA表达量明显减少(P<0.01),BLT1 mRNA表达与对照组无明显差异,p-Erk表达量与MK886组比较明显增加(P<0.01),MK886+80?mol/L Celecoxib作用下,BLT1 mRNA、VEGF mRNA表达量明显减少(P<0.01),p-Erk表达量与对照组无明显差异.结论:花生四烯酸的两条代谢途径均与胰腺癌的发生及增殖均有密切关系,而抑制5-脂氧合酶(5-lipoxygenase)途径较环氧化酶2(cyclooxygenase 2)途径相比,抑制肿瘤细胞增殖作用更强.
文摘目的探讨细丝蛋白C(filamin C,FLNC)对前列腺癌细胞迁移和侵袭的影响。方法通过免疫组织化学分析了30例前列腺癌患者的癌组织及配对癌旁组织中FLNC的表达。根据FLNC蛋白的表达水平将30例前列腺癌患者分为FLNC低表达组(0~3分,n=9)和FLNC高表达组(4~12分,n=21),采用Kaplan-Meier法进行生存分析。对人前列腺癌细胞系(PC-3)转染抑制FLNC表达的小干扰RNA(siRNA-FLNC组)和阴性对照(siRNA-NC组),未转染的细胞作为对照组。通过MTT法检测细胞活力,Transwell实验进行迁移和侵袭分析。通过qRT-PCR或Western blot检测细胞中FLNC、Eotaxin-1、CCR3、p-ERK1/2、t-ERK1/2和MMP-3的表达。结果与癌旁组织相比,前列腺癌组织中FLNC的染色评分升高(1.43±0.86 vs 5.63±2.91,P<0.001)。FLNC低表达组和高表达组患者的TNM分期、淋巴结转移和生存时间差异有统计学意义(P<0.05)。与对照组相比,siRNA-FLNC组PC-3细胞中FLNC的mRNA和蛋白水平均降低(P<0.05),细胞活力降低(P<0.05),细胞迁移和侵袭数量降低(P<0.05)。与对照组相比,siRNA-FLNC组Eotaxin-1、CCR3、p-ERK1/2和MMP-3的蛋白相对表达量均降低(P<0.05),而t-ERK1/2的蛋白相对表达量无明显变化(P>0.05)。结论FLNC在前列腺癌组织中高表达并且与患者预后相关。沉默FLNC可能通过抑制Eotaxin-1-CCR3-ERK1/2-MMP-3通路降低前列腺癌细胞的迁移和侵袭能力。
