目的探究负载膜联蛋白A2(ANXA2)的骨髓间充质干细胞(BMSC)来源的外泌体(Exo-ANXA2)对前列腺癌细胞增殖、侵袭与迁移以及前列腺癌移植瘤生长的影响,并研究巨噬细胞是否参与该过程。方法分离与培养BALB/c裸鼠BMSC,采用负载ANXA2的慢病毒...目的探究负载膜联蛋白A2(ANXA2)的骨髓间充质干细胞(BMSC)来源的外泌体(Exo-ANXA2)对前列腺癌细胞增殖、侵袭与迁移以及前列腺癌移植瘤生长的影响,并研究巨噬细胞是否参与该过程。方法分离与培养BALB/c裸鼠BMSC,采用负载ANXA2的慢病毒质粒感染BMSC,并分离外泌体;加入外泌体处理THP-1巨噬细胞,ELISA检测细胞上清培养液肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-6和IL-10的水平;将外泌体处理的巨噬细胞与前列腺癌细胞共培养后,CCK-8法检测细胞增殖活性,Transwell^(TM)小室检测细胞侵袭与迁移。通过注射PC-3人前列腺癌细胞构建前列腺癌裸鼠移植瘤模型,将建模后的裸鼠随机分为对照组和实验组,每组8只,实验组裸鼠尾静脉注射1 mL Exo-ANXA2,对照组注射等量PBS,于注射后第0、3、6、9、12、15、18、21天,使用游标卡尺测量计算肿瘤体积,21 d处死裸鼠并测量肿瘤组织质量,免疫组织化学染色法检测肿瘤组织中抗原KI-67(ki67)和CD163表达。结果骨髓分离的细胞表面CD90和CD44呈高表达,CD34和CD45呈低表达,且成骨成脂分化能力较强,成功获得BMSC;负载ANXA2的慢病毒质粒感染后,BMSC中有较强的绿色荧光蛋白表达,并成功分离到Exo-ANXA2;经Exo-ANXA2处理后,THP-1细胞中TNF-α和IL-6的水平显著升高而IL-10和IL-13的水平显著下降;Exo-ANXA2处理巨噬细胞后,显著抑制Exo-ANXA2促进PC-3细胞增殖活性、侵袭与迁移的作用;经过给予前列腺癌移植瘤裸鼠注射Exo-ANXA2后,裸鼠在第6、9、12、15、18、21天的肿瘤组织体积显著减小,21 d时裸鼠肿瘤体质量也显著减少,此外,肿瘤组织内ki67和CD163的阳性表达率均显著降低。结论Exo-ANXA2能够抑制前列腺癌细胞增殖、侵袭与迁移,并抑制裸鼠前列腺癌移植瘤生长,M2型巨噬细胞减少。展开更多
Germinal matrix hemorrhage is one of the leading causes of morbidity,mortality,and acquired infantile hydrocephalus in preterm infants in the United States,with little progress made in its clinical management.Blood cl...Germinal matrix hemorrhage is one of the leading causes of morbidity,mortality,and acquired infantile hydrocephalus in preterm infants in the United States,with little progress made in its clinical management.Blood clots have been shown to elicit secondary brain injury after germinal matrix hemorrhage,by disrupting normal cerebrospinal fluid circulation and absorption after germinal matrix hemorrhage causing post-hemorrhagic hydrocephalus development.Current evidence suggests that rapid hematoma resolution is necessary to improve neurological outcomes after hemorrhagic stroke.Various articles have demonstrated the beneficial effects of stimulating the polarization of microglia cells into the M2 phenotype,as it has been suggested that they play an essential role in the rapid phagocytosis of the blood clot after hemorrhagic models of stroke.N-formyl peptide receptor 2(FPR2),a G-protein-coupled receptor,has been shown to be neuroprotective after stroke.FPR2 activation has been associated with the upregulation of phagocytic macrophage clearance,yet its mechanism has not been fully explored.Recent literature suggests that FPR2 may play a role in the stimulation of scavenger receptor CD36.Scavenger receptor CD36 plays a vital role in microglia phagocytic blood clot clearance after germinal matrix hemorrhage.FPR2 has been shown to phosphorylate extracellular-signal-regulated kinase 1/2(ERK1/2),which then promotes the transcription of the dual-specificity protein phosphatase 1(DUSP1)gene.In this review,we present an intrinsic outline of the main components involved in FPR2 stimulation and hematoma resolution after germinal matrix hemorrhage.展开更多
文摘目的探究负载膜联蛋白A2(ANXA2)的骨髓间充质干细胞(BMSC)来源的外泌体(Exo-ANXA2)对前列腺癌细胞增殖、侵袭与迁移以及前列腺癌移植瘤生长的影响,并研究巨噬细胞是否参与该过程。方法分离与培养BALB/c裸鼠BMSC,采用负载ANXA2的慢病毒质粒感染BMSC,并分离外泌体;加入外泌体处理THP-1巨噬细胞,ELISA检测细胞上清培养液肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-6和IL-10的水平;将外泌体处理的巨噬细胞与前列腺癌细胞共培养后,CCK-8法检测细胞增殖活性,Transwell^(TM)小室检测细胞侵袭与迁移。通过注射PC-3人前列腺癌细胞构建前列腺癌裸鼠移植瘤模型,将建模后的裸鼠随机分为对照组和实验组,每组8只,实验组裸鼠尾静脉注射1 mL Exo-ANXA2,对照组注射等量PBS,于注射后第0、3、6、9、12、15、18、21天,使用游标卡尺测量计算肿瘤体积,21 d处死裸鼠并测量肿瘤组织质量,免疫组织化学染色法检测肿瘤组织中抗原KI-67(ki67)和CD163表达。结果骨髓分离的细胞表面CD90和CD44呈高表达,CD34和CD45呈低表达,且成骨成脂分化能力较强,成功获得BMSC;负载ANXA2的慢病毒质粒感染后,BMSC中有较强的绿色荧光蛋白表达,并成功分离到Exo-ANXA2;经Exo-ANXA2处理后,THP-1细胞中TNF-α和IL-6的水平显著升高而IL-10和IL-13的水平显著下降;Exo-ANXA2处理巨噬细胞后,显著抑制Exo-ANXA2促进PC-3细胞增殖活性、侵袭与迁移的作用;经过给予前列腺癌移植瘤裸鼠注射Exo-ANXA2后,裸鼠在第6、9、12、15、18、21天的肿瘤组织体积显著减小,21 d时裸鼠肿瘤体质量也显著减少,此外,肿瘤组织内ki67和CD163的阳性表达率均显著降低。结论Exo-ANXA2能够抑制前列腺癌细胞增殖、侵袭与迁移,并抑制裸鼠前列腺癌移植瘤生长,M2型巨噬细胞减少。
基金supported in part by the National Institutes of Health grant 5R01NS117364-02(to JT)。
文摘Germinal matrix hemorrhage is one of the leading causes of morbidity,mortality,and acquired infantile hydrocephalus in preterm infants in the United States,with little progress made in its clinical management.Blood clots have been shown to elicit secondary brain injury after germinal matrix hemorrhage,by disrupting normal cerebrospinal fluid circulation and absorption after germinal matrix hemorrhage causing post-hemorrhagic hydrocephalus development.Current evidence suggests that rapid hematoma resolution is necessary to improve neurological outcomes after hemorrhagic stroke.Various articles have demonstrated the beneficial effects of stimulating the polarization of microglia cells into the M2 phenotype,as it has been suggested that they play an essential role in the rapid phagocytosis of the blood clot after hemorrhagic models of stroke.N-formyl peptide receptor 2(FPR2),a G-protein-coupled receptor,has been shown to be neuroprotective after stroke.FPR2 activation has been associated with the upregulation of phagocytic macrophage clearance,yet its mechanism has not been fully explored.Recent literature suggests that FPR2 may play a role in the stimulation of scavenger receptor CD36.Scavenger receptor CD36 plays a vital role in microglia phagocytic blood clot clearance after germinal matrix hemorrhage.FPR2 has been shown to phosphorylate extracellular-signal-regulated kinase 1/2(ERK1/2),which then promotes the transcription of the dual-specificity protein phosphatase 1(DUSP1)gene.In this review,we present an intrinsic outline of the main components involved in FPR2 stimulation and hematoma resolution after germinal matrix hemorrhage.
