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Anti-proliferative effect of Annona extracts on breast cancer cells. 番荔枝提取物对乳腺癌细胞的抗增殖作用。
M. Veisaga, M. Ahumada, Stacy Soriano, L. Acuña, W. Zhang, Ivy Leung, Robert P. Barnum, Manuel A. BARBIERI
BackgorundFruits and seed extracts of Annona montana have significant cytotoxic potential in several cancer cells. This study evaluates the effect of A. montana leaves hexane extract on several signaling cascades and gene expression in metastatic breast cancer cells upon insulin-like growth factor-1 (IGF-1) stimulation.MethodsMTT assay was performed to determine the proliferation of cancer cells. Propidium iodide staining and flow cytometry analysis of Annexin V binding was utilized to measure the progression of the cell cycle and the induction of apoptosis. Protein expression and phosphorylation were determined by western blotting analysis to examine the underlying cellular mechanism triggered upon treatment with A. montana leaves hexane extract.ResultsA. montana leaves hexane (sub-fraction V) blocked the constitutive stimulation of the PI3K/mTOR signaling pathways. This inhibitory effect was associated with apoptosis induction as evidenced by the positivity with Annexin V and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNNEL) staining, activation of caspase-3, and cleavage of PPAR. It also limited the expression of various downstream genes that regulate proliferation, survival, metastasis, and angiogenesis (i.e., cyclin D1, survivin, COX-2, and VEGF). It increased the expression of p53 and p21. Interestingly, we also observed that this extract blocked the activation of AKT and ERK without affecting the phosphorylation of the IGF-1 receptor and activation of Ras upon IGF-1 stimulation.ConclusionOur study indicates that A. montana leaves (sub-fraction V) extract exhibits a selective anti-proliferative and proapoptotic effect on the metastatic MDA-MB-231 breast cancer cells through the involvement of PI3K/AKT/mTOR/S6K1 pathways.
背景蒙大拿州番荔枝果实和种子提取物对几种癌细胞具有显著的细胞毒性。本研究评估了蒙大拿树叶己烷提取物对转移性乳腺癌细胞中胰岛素样生长因子-1 (IGF-1)刺激的信号级联反应和基因表达的影响。方法采用smtt法检测肿瘤细胞增殖情况。采用碘化丙啶染色和Annexin V结合的流式细胞术检测细胞周期的进展和诱导凋亡的情况。western blotting检测蛋白表达和磷酸化水平,探讨蒙大拿叶己烷提取物诱发的潜在细胞机制。蒙大拿叶己烷(亚组分V)阻断了PI3K/mTOR信号通路的组成性刺激。Annexin V和末端脱氧核苷酸转移酶dUTP缺口末端标记(TUNNEL)染色阳性,caspase-3激活,PPAR裂解,证明了这种抑制作用与细胞凋亡诱导有关。它还限制了调节增殖、生存、转移和血管生成的各种下游基因(即cyclin D1、survivin、COX-2和VEGF)的表达。增加p53和p21的表达。有趣的是,我们还观察到该提取物阻断了AKT和ERK的激活,而不影响IGF-1受体的磷酸化和IGF-1刺激后Ras的激活。结论蒙大纳叶提取物通过参与PI3K/AKT/mTOR/S6K1通路,对转移性乳腺癌MDA-MB-231细胞具有选择性的抗增殖和促凋亡作用。
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引用次数: 0
Recent biomedical advances enabled by HaloTag technology HaloTag技术使生物医学取得了最新进展
Weiyu Chen, Muhsin H Younis, Zhongkuo Zhao, W. Cai
The knowledge of interactions among functional proteins helps researchers understand disease mechanisms and design potential strategies for treatment. As a general approach, the fluorescent and affinity tags were employed for exploring this field by labeling the Protein of Interest (POI). However, the autofluorescence and weak binding strength significantly reduce the accuracy and specificity of these tags. Conversely, HaloTag, a novel self-labeling enzyme (SLE) tag, could quickly form a covalent bond with its ligand, enabling fast and specific labeling of POI. These desirable features greatly increase the accuracy and specificity, making the HaloTag a valuable system for various applications ranging from imaging to immobilization of POI. Notably, the HaloTag technique has already been successfully employed in a series of studies with excellent efficiency. In this review, we summarize the development of HaloTag and recent advanced investigations associated with HaloTag, including in vitro imaging (e.g., POI imaging, cellular condition monitoring, microorganism imaging, system development), in vivo imaging, biomolecule immobilization (e.g., POI collection, protein/nuclear acid interaction and protein structure analysis), targeted degradation (e.g., L-AdPROM), and more. We also present a systematic discussion regarding the future direction and challenges of the HaloTag technique.
功能蛋白之间相互作用的知识有助于研究人员了解疾病机制并设计潜在的治疗策略。一般采用荧光和亲和标签标记感兴趣蛋白(POI)来探索这一领域。然而,自身荧光和弱结合强度显著降低了这些标签的准确性和特异性。相反,HaloTag是一种新型的自标记酶(SLE)标签,它可以快速地与其配体形成共价键,从而实现对POI的快速和特异性标记。这些理想的特性大大提高了准确性和特异性,使HaloTag成为从成像到POI固定的各种应用的有价值的系统。值得注意的是,HaloTag技术已经成功地应用于一系列高效的研究中。在这篇综述中,我们总结了HaloTag的发展以及与HaloTag相关的最新研究进展,包括体外成像(例如POI成像、细胞状态监测、微生物成像、系统开发)、体内成像、生物分子固定化(例如POI收集、蛋白质/核酸相互作用和蛋白质结构分析)、靶向降解(例如L-AdPROM)等。我们还对HaloTag技术的未来方向和挑战进行了系统的讨论。
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引用次数: 3
Synergy of single-cell sequencing analyses and in vivo lineage-tracing approaches: A new opportunity for stem cell biology 单细胞测序分析和体内谱系追踪方法的协同作用:干细胞生物学的新机遇
Yuki Matsushita, W. Ono, N. Ono
Single-cell sequencing technologies have rapidly progressed in recent years, and been applied to characterize stem cells in a number of organs. Somatic (postnatal) stem cells are generally identified using combinations of cell surface markers and transcription factors. However, it has been challenging to define micro-heterogeneity within “stem cell” populations, each of which stands at a different level of differentiation. As stem cells become defined at a single-cell level, their differentiation path becomes clearly defined. Here, this viewpoint discusses the potential synergy of single-cell sequencing analyses with in vivo lineage-tracing approaches, with an emphasis on practical considerations in stem cell biology.
单细胞测序技术近年来发展迅速,已被应用于许多器官的干细胞鉴定。体细胞(出生后)干细胞通常使用细胞表面标记和转录因子的组合来鉴定。然而,定义“干细胞”群体中的微观异质性一直具有挑战性,每个群体都处于不同的分化水平。随着干细胞在单细胞水平上被定义,它们的分化路径也变得清晰。在这里,这一观点讨论了单细胞测序分析与体内谱系追踪方法的潜在协同作用,重点是干细胞生物学中的实际考虑。
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引用次数: 2
New paradigms in regenerative engineering: Emerging role of extracellular vesicles paired with instructive biomaterials 再生工程的新范例:细胞外囊泡与指导性生物材料配对的新作用
W. Swanson, Yuji Mishina
Mesenchymal stem cells (MSCs) have long been regarded as critical components of regenerative medicine strategies, given their multipotency and persistence in a variety of tissues. Recently, the specific role of MSCs in mediating regenerative outcomes has been attributed (in part) to secreted factors from transplanted cells, namely extracellular vesicles. This viewpoint manuscript highlights the promise of cell-derived extracellular vesicles as agents of regeneration, enhanced by synergy with appropriate biomaterials platforms. Extracellular vesicles are a potentially interesting regenerative tool to enhance the synergy between MSCs and biomaterials. As a result, we believe these technologies will improve patient outcomes through efficient therapeutic strategies resulting in predictable patient outcomes.
间充质干细胞(MSCs)长期以来被认为是再生医学策略的关键组成部分,因为它们具有多能性和在多种组织中的持久性。最近,MSCs在介导再生结果中的特殊作用(部分)归因于移植细胞的分泌因子,即细胞外囊泡。这一观点手稿强调了细胞来源的细胞外囊泡作为再生剂的前景,通过与适当的生物材料平台协同作用增强。细胞外囊泡是一种潜在的有趣的再生工具,可以增强间充质干细胞和生物材料之间的协同作用。因此,我们相信这些技术将通过有效的治疗策略改善患者的治疗效果,从而产生可预测的患者结果。
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引用次数: 4
Stem cells in intervertebral disc regeneration–more talk than action? 干细胞在椎间盘再生中的应用——空谈少行动?
P. Kraus, Ankit Samanta, Sina Lufkin, T. Lufkin
Pain and lifestyle changes are common consequences of intervertebral disc degeneration (IVDD) and affect a large part of the aging population. The stemness of cells is exploited in the field of regenerative medicine as key to treat degenerative diseases. Transplanted cells however often face delivery and survival challenges, especially in tissues with a naturally harsh microniche environment such as the intervertebral disc. Recent interest in the secretome of stem cells, especially cargo protected from microniche-related decay as frequently present in degenerating tissues, provides new means of rejuvenating ailing cells and tissues. Exosomes, a type of extracellular vesicles with purposeful cargo gained particular interest in conveying stem cell related attributes of rejuvenation, which will be discussed here in the context of IVDD.
疼痛和生活方式的改变是椎间盘退变(IVDD)的常见后果,影响了很大一部分老龄化人口。在再生医学领域,细胞的干细胞性是治疗退行性疾病的关键。然而,移植细胞往往面临递送和存活的挑战,特别是在具有自然恶劣微微环境的组织中,如椎间盘。最近对干细胞分泌组的研究,特别是对经常出现在退化组织中的微颗粒相关衰变的保护,提供了使患病细胞和组织恢复活力的新方法。外泌体是一种具有目的货物的细胞外囊泡,在传递干细胞相关的年轻化属性方面获得了特别的兴趣,这将在IVDD的背景下进行讨论。
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引用次数: 3
Proteogenomics for pediatric brain cancer 儿童脑癌的蛋白质基因组学研究
M. Simonian
Pediatric central nervous system tumors are the most common tumors in children, it constitute 15%–20% of all malignancies in children and are the leading cause of cancer related deaths in children. Proteogenomics is an emerging field of biological research that utilizes a combination of proteomics, genomics, and transcriptomics to aid in the discovery and identification of biomarkers for diagnosis and therapeutic purposes. Integrative proteogenomics analysis of pediatric tumors identified underlying biological processes and potential treatments as well as the functional effects of somatic mutations and copy number variation driving tumorigenesis.
小儿中枢神经系统肿瘤是儿童中最常见的肿瘤,占儿童所有恶性肿瘤的15%-20%,是儿童癌症相关死亡的主要原因。蛋白质基因组学是生物学研究的一个新兴领域,它利用蛋白质组学、基因组学和转录组学的结合来帮助发现和鉴定用于诊断和治疗目的的生物标志物。儿童肿瘤的综合蛋白质基因组学分析确定了潜在的生物学过程和潜在的治疗方法,以及体细胞突变和拷贝数变异驱动肿瘤发生的功能影响。
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引用次数: 0
Aneuploidy: An Opportunity Within Single-Cell RNA Sequencing Analysis 非整倍体:单细胞RNA测序分析的机会
J. Delaney
Single-cell sequencing data has transformed the understanding of biological heterogeneity. While many flavors of single-cell sequencing have been developed, single-cell RNA sequencing (scRNA-seq) is currently the most prolific form in published literature. Bioinformatic analysis of differential biology within the population of cells studied relies on inferences and grouping of cells due to the spotty nature of data within individual cell scRNA-seq gene counts. One biologically relevant variable is readily inferred from scRNA-seq gene count tables regardless of individual gene representation within single cells: aneuploidy. Since hundreds of genes are present on chromosome arms, high-quality inferences of aneuploidy can be made from scRNA-seq datasets. This viewpoint summarizes how utilization of these bioinformatic pipelines can benefit scRNA-seq studies, particularly in oncology wherein aneuploidy is both rampant and a hallmark of the studied disease. Awareness and use of these analytical pipelines will improve each field’s ability to understand the studied diseases. Authors are encouraged to attempt these aneuploid analyses when reporting scRNA-seq data, much like copy-number variants are commonly reported in bulk genome sequencing data.
单细胞测序数据已经改变了对生物异质性的理解。虽然已经开发了许多单细胞测序方法,但单细胞RNA测序(scRNA-seq)是目前已发表文献中最丰富的形式。由于单个细胞scRNA-seq基因计数数据的不均匀性,对所研究细胞群体内差异生物学的生物信息学分析依赖于对细胞的推断和分组。从scRNA-seq基因计数表中可以很容易地推断出一个生物学上相关的变量:非整倍性,而不管单个基因在单细胞中的表现如何。由于染色体臂上存在数百个基因,因此可以从scRNA-seq数据集获得高质量的非整倍性推断。这一观点总结了利用这些生物信息管道如何使scRNA-seq研究受益,特别是在非整倍体既猖獗又被研究疾病的肿瘤中。了解和使用这些分析管道将提高每个领域了解所研究疾病的能力。在报告scRNA-seq数据时,鼓励作者尝试这些非整倍体分析,就像在大量基因组测序数据中通常报告的拷贝数变异一样。
{"title":"Aneuploidy: An Opportunity Within Single-Cell RNA Sequencing Analysis","authors":"J. Delaney","doi":"10.32604/biocell.2021.017296","DOIUrl":"https://doi.org/10.32604/biocell.2021.017296","url":null,"abstract":"Single-cell sequencing data has transformed the understanding of biological heterogeneity. While many flavors of single-cell sequencing have been developed, single-cell RNA sequencing (scRNA-seq) is currently the most prolific form in published literature. Bioinformatic analysis of differential biology within the population of cells studied relies on inferences and grouping of cells due to the spotty nature of data within individual cell scRNA-seq gene counts. One biologically relevant variable is readily inferred from scRNA-seq gene count tables regardless of individual gene representation within single cells: aneuploidy. Since hundreds of genes are present on chromosome arms, high-quality inferences of aneuploidy can be made from scRNA-seq datasets. This viewpoint summarizes how utilization of these bioinformatic pipelines can benefit scRNA-seq studies, particularly in oncology wherein aneuploidy is both rampant and a hallmark of the studied disease. Awareness and use of these analytical pipelines will improve each field’s ability to understand the studied diseases. Authors are encouraged to attempt these aneuploid analyses when reporting scRNA-seq data, much like copy-number variants are commonly reported in bulk genome sequencing data.","PeriodicalId":342778,"journal":{"name":"Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al","volume":"40 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121672343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 1
Post-treatment with plant extracts used in Brazilian folk medicine caused a partial reversal of the antiproliferative effect of glyphosate in the Allium cepa test. 用巴西民间医学中使用的植物提取物进行后处理,导致葱试验中草甘膦的抗增殖作用部分逆转。
V. Frescura, A. W. Kuhn, H. D. Laughinghouse, J. T. Paranhos, S. Tedesco
Species of the genus Psychotria are used for multiple purposes in Brazilian folk medicine, either as water infusions, baths or poultices. This study was aimed to evaluate the genotoxic and antiproliferative effects of infusions of Psychotria brachypoda and P. birotula on the Allium cepa test. Exposure to distilled water was used as a negative control, while exposure to glyphosate was used as a positive control. The interaction of extracts (as a post-treatment) with the effects of glyphosate was also studied. Results showed that glyphosate and the extracts of both P. brachypoda and P. birotula reduced the mitotic index as compared with the negative control (distilled water). Surprisingly, however, both extracts from P. brachypoda and P. birotula caused a partial reversal of the antiproliferative effect of glyphosate when used as a post-treatment. Glyphosate also induced the highest number of cells with chromosomal alterations, which was followed by that of P. birotula extracts. However, the extracts from P. brachypoda did not show any significant genotoxic effect. Post-treatment of glyphosate-treated samples with distilled water allowed a partial recovery of the genotoxic effect of glyphosate, and some of the Psychotria extracts also did so. Notably, post-treatment of glyphosate-treated samples with P. brachypoda extracts induced a statistically significant apoptotic effect. It is concluded that P. brachypoda extracts show antiproliferative effects and are not genotoxic, while extracts of P. birotula show a less potent antiproliferative effect and may induce chromosomal abnormalities. The finding of a partial reversion of the effects of glyphosate by a post-treatment with extracts from both plants should be followed up.
在巴西民间医学中,精神病属的物种被用于多种用途,无论是作为水输液,沐浴还是膏药。本研究旨在探讨短足精神草和小圆藤对大蒜试验的遗传毒性和抗增殖作用。暴露于蒸馏水被用作阴性对照,而暴露于草甘膦被用作阳性对照。还研究了提取物(作为后处理)与草甘膦作用的相互作用。结果表明,与阴性对照(蒸馏水)相比,草甘膦与短足假虾和小圆尾假虾提取物均降低了有丝分裂指数。然而,令人惊讶的是,当用作后处理时,短足假蓟马和小轮假蓟马的提取物都能部分逆转草甘膦的抗增殖作用。草甘膦诱导的染色体改变细胞数量最多,其次是小轮草提取物。然而,短足草提取物没有表现出明显的遗传毒性作用。用蒸馏水对经过草甘膦处理的样本进行后处理,可以部分恢复草甘膦的基因毒性作用,一些精神科提取物也有这种作用。值得注意的是,草甘膦处理后的样品与短足草提取物诱导了统计学上显著的凋亡效应。由此可见,短足棘提取物具有抗增殖作用,且不具有遗传毒性,而小轮棘提取物的抗增殖作用较弱,且可能引起染色体异常。用这两种植物的提取物进行后处理后,草甘膦的作用部分恢复的发现应予以跟进。
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引用次数: 14
Confocal laser scanning microscopy and immunohistochemistry of cerebellar Lugaro cells. 小脑Lugaro细胞的共聚焦激光扫描显微镜和免疫组织化学。
O. Castejón
The present paper shows by means of confocal laser scanning microscopy the immunoreactivity of rat cerebellar Lugaro cells for calbindin, synapsin-I, PSD-95, GluR1, CaMKII alpha, and N-cadherin. Lugaro cells were easily characterized by their location beneath Purkinje cells. Calbindin revealed immunoreactivity in the cell body, and the axonal and dendritic processes. Synapsin-I labelled the presynaptic endings on Lugaro cells. Synapsin-I and PSD-95 immunoreactivity demonstrated the localization of presynaptic and postsynaptic endings surrounding cell soma, corresponding to afferent extrinsic and intrinsic cerebellar fibers. GluR1 immunoreactivity of the soma and cell processes indicates that Lugaro cells have functional ionotropic glutamate receptors that regulate calcium levels. CaMKII alpha immunoreactivity of Lugaro cell soma and processes suggest its participation as a molecular switch for long-term information storage, and serving as a molecular basis of long-term synaptic memory. N-cadherin immunoreactivity was correlated with somato-somatic and somato-dendritic junctions between Lugaro cells and their synaptic connections.
本文用激光共聚焦扫描显微镜观察了大鼠小脑Lugaro细胞对calbindin、synapsin-I、PSD-95、GluR1、CaMKII α和N-cadherin的免疫反应性。卢加罗细胞的特征在于它们位于浦肯野细胞之下。Calbindin在细胞体、轴突和树突中显示免疫反应性。突触素- 1标记Lugaro细胞的突触前末梢。突触素- 1和PSD-95的免疫反应性表明突触前和突触后终末在细胞体周围的定位,对应于传入的外源性和内源性小脑纤维。胞体和细胞过程的GluR1免疫反应性表明Lugaro细胞具有调节钙水平的功能性离子性谷氨酸受体。Lugaro细胞体细胞和过程的CaMKII α免疫反应性表明其参与长期信息存储的分子开关,并作为长期突触记忆的分子基础。n -钙粘蛋白免疫反应性与Lugaro细胞之间的体-体和体-树突连接及其突触连接相关。
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引用次数: 3
Creatine supplementation in trained rats causes changes in myenteric neurons and intestinal wall morphometry. 在训练大鼠中补充肌酸可引起肌神经细胞和肠壁形态的改变。
S. M. D. de Moraes, Thais Andréia Brogio, J. Zanoni, M. Zapater, S. B. Peres, L. Hernandes
Creatine is widely used by athletes as an ergogenic resource. The aim of this study was to evaluate the influence of creatine supplementation on the duodenum of rats submitted to physical training. The number and myenteric neuronal cell bodies as well mucosal and muscular tunic morphometry were evaluated. Control animals received a standard chow for 8 weeks, and the treated ones received the standard chow for 4 weeks and were later fed with the same chow but added with 2% creatine. Animals were divided in groups: sedentary, sedentary supplemented with creatine, trained and trained supplemented with creatine. The training consisted in treadmill running for 8 weeks. Duodenal samples were either processed for whole mount preparations or for paraffin embedding and hematoxylin-eosin staining for histological and morphometric studies of the mucosa, the muscular tunic and myenteric neurons. It was observed that neither creatine nor physical training alone promoted alterations in muscular tunic thickness, villus height or crypts depth, however, a reduction in these parameters was observed when both were associated. The number of myenteric neurons was unchanged, but the neuronal cell body area was reduced in trained animals but not when training and creatine was associated, suggesting a neuroprotector role of this substance.
肌酸是运动员广泛使用的一种能量来源。本研究的目的是评价补充肌酸对体育训练大鼠十二指肠的影响。测定了肌肠神经元胞体的数量及粘膜和肌束形态。对照组饲喂标准饲料8周,治疗组饲喂标准饲料4周,再饲喂添加2%肌酸的标准饲料。动物被分成几组:久坐的,久坐的补充肌酸,训练的和训练的补充肌酸。训练包括在跑步机上跑步8周。对十二指肠标本进行全载或石蜡包埋和苏木精-伊红染色,对粘膜、肌束膜和肌肠神经元进行组织学和形态计量学研究。我们观察到,肌酸和体能训练都不能单独促进肌肉束膜厚度、绒毛高度或隐窝深度的改变,然而,当两者都相关时,可以观察到这些参数的减少。在训练的动物中,肌肠神经元的数量没有变化,但神经元细胞的体面积减少了,但当训练和肌酸相关联时,这种情况没有发生,这表明这种物质具有神经保护作用。
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引用次数: 3
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Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al
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