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Rho GTPases与肿瘤侵袭转移相关性的研究进展

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国际遗传学杂志 2016年第1期39卷 15-21页

作者：庞博孙冬琳孙海明金焰傅松滨哈尔滨医科大学医学遗传学研究室

Rho GTPases属小分子量gtpases,作为信号传导通路中重要的"分子开关",调控细胞形态、骨架重建、迁移及细胞周期与凋亡等诸多过程。近来的研究表明,Rho GTPases家族成员在多种恶性肿瘤中异常表达,并通过调控上皮细胞极性丧失、细胞-细胞... 详细信息

Rho GTPases属小分子量gtpases,作为信号传导通路中重要的"分子开关",调控细胞形态、骨架重建、迁移及细胞周期与凋亡等诸多过程。近来的研究表明,Rho GTPases家族成员在多种恶性肿瘤中异常表达,并通过调控上皮细胞极性丧失、细胞-细胞间的连接及黏附、细胞骨架重组和迁移运动、细胞外基质的重塑、细胞进入血管淋巴管的能力等方面参与肿瘤细胞的侵袭转移。深入研究Rho GTPases在肿瘤侵袭转移中的作用意义重大,rho GT—Pases将有望成为判断肿瘤侵袭转移性发展的新指标,并为肿瘤治疗提供新靶点、新方向。

关键词： Rho GTPases 肿瘤侵袭转移

The effects of 8 weeks treadmill exercise on the Rho GTPases in rat special brain regions

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The effects of 8 weeks treadmill exercise on the Rho GTPases...

The role of rho GTPase family in cochlear hair cells and hearing

中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Yan LI,Li ZHAO~*,Li-Jing GONG Beijing Sport University,Beijing 100084,China

＜正＞Objective Recently numerous of epidemiological data have proven that physical activity or regular exercise beneficially affects brain function and increases neuronal development,but the mechanisms of exercise has not been clear.Rho GTPases act as the central regulators of cytoskeleton and function as binary molecular switches. In this family,rhoA,Racl and Cdc42 have been characterized most extensively.Recently,the role of Rho GTPases in neuronal development has been studied.Therefore,the aim of this study was to investigate the effects of 8 weeks treadmill exercise on the Rho GTPases in the rat special brain regions.Methods 3 months male Wistar rats were randomly assigned to either a sedentary or a treadmill exercise group.Exercised rats were familiarized to treadmill exercise for 5 days,then ran 30 min/day at 10m/min on a 5%slope for the first 2 weeks.The running speed and duration of the exercise were gradually increased,up to 60 min/day at 22 m/min on a 10%slope for 6 weeks.All rats were killed 48 h after the last exercise session.The levels of synaptophysin in the cortex and hippocampus were separately detected with the ICH and IF method,which has been widely used to assess synapse density.Furthermore, the protein levels of rhoA,Rac1 and Cdc42 were measured using Western blotting and the mRNA levels were measured using real time PCR.Data were analyzed using one-way analysis of variance（ANOVA）.The level of statistical significance was set at P＜0.05.Results（1） The increase of synapse density appeared in both cortex（P＜0.05） and hippocampus（P＜0.05） in exercised groups compared with sedentary groups.（2） Exercise decreased the protein levels of rhoA in both cortex（P＜0.01） and hippocampus（P＜0.01）,at the same time increased the mRNA levels in both cortex（P＜0.001） and hippocampus（PO.001）.（3） Exercise increased the protein levels of Rac1 in both cortex （P＜0.01） and hippocampus（P＜0.01）.Exercise increased the mRNA level of Racl in cortex（P＜0.01） but decreased the mRNA level in hippocampus（P＜0.01）.（4） Exercise increase

关键词： exercise Rho GTPases neuronal development

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Neural Regeneration Research 2023年第10期18卷 2167-2172页

作者： Yu-Bei Dai Xiang Gao Dong Liu Jie Gong School of Life Science Nantong Laboratory of Development and DiseasesKey Laboratory of Neuroregeneration of Jiangsu and Ministry of EducationCo-innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina

Rho GTPases are essential regulators of the actin cytoskeleton.They are involved in various physiological and biochemical processes such as the regulation of cytoskeleton dynamics,development,proliferation,survival,and regeneration.During the development of cochlear hair cells,Rho GTPases are activated by various extracellular signals through membrane receptors to further stimulate multiple downstream effectors.Specifically,rhoA,Cdc42,and Rac1,members of the classical subfamily of the rho GTPase family,regulate the development and maintenance of cilia by inducing the polymerization of actin monomers and stabilizing actin filaments.In addition,they also regulate the normal morphology orientation of ciliary bundles in auditory hair cells,which is an important element of cell polarity regulation.Moreover,the actin-related pathways mediated by rhoA,Cdc42,and Rac1 also play a role in the motility of outer hair cells,indicating that the function of Rho GTPases is crucial in the highly polar auditory sensory system.In this review,we focus on the expression of rhoA,Cdc42,and Rac1 in cochlear hair cells and how these small molecules participate in ciliary bundle morphogenesis and cochlear hair cell movement.We also discuss the progress of current research investigating the use of these small molecules as drug targets for deafness treatment.

关键词： actin assembly auditory sensory neurons cell polarity cell proliferation electromotility hair cell hearing loss morphogenesis Rho GTPases stereocilia

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A positive feedback circuit for ROP-mediated polar growth

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Molecular Plant 2021年第3期14卷 395-410页

作者： En Li Yu-Ling Zhang Xuelian Shi Han Li Xuefeng Yuan Sha Li Yan Zhang State Key Laboratory of Crop Biology College of Life SciencesShandong Agricultural UniversityTai'an 271018China Shandong Province Key Laboratory of Agricultural Microbiology College of Plant ProtectionShandong Agricultural UniversityTai'an 271018China

Tip growth is a special type of polarized growth in which a single and unique polarization site is established and maintained.rho of Plants(ROP)proteins,which represent the only class of Rho GTPases in plants,regulate tip growth.The dynamic and asymmetric distribution of ROPs is critical for the establishment and maintenance of tip growth,and requires at least one positive feedback loop,which is still elusive.Here,we report a positive feedback circuit essential for tip growth of root hairs,in which ROPs,ROP activators and effectors,and AGC1.5 subfamily kinases are interconnected by sequential oligomerization and phosphorylation.AGC1.5 subfamily kinases interact with and phosphorylate two guanine nucleotide exchange factors(GEFs)of ROPs,RopGEF4 and RopGEF10.They also interact with two ROP effectors,ICR2/RIP3 and MIDD1/RIP4,which bridge active ROPs with AGC1.5.Functional loss of the AGC1.5 subfamily kinases or ICR2 and MIDD1 compromised root hair growth due to reduced ROP signaling.We found that asymmetric targeting of RopGEF4 and RopGEF10 is controlled by AGC1.5-dependent phosphorylation.Interestingly,we discovered that the ROP effectors recruit AGC1.5 to active ROP domains at the plasma membrane during root hair growth and are critical for AGC1.5-dependent phosphorylation of RopGEFs.Given the large number of AGC kinases in plants,this positive feedback circuit may be a universal theme for plant cell polar growth.

关键词： Rho GTPases AGC kinase membrane recruitment phosphorylation asymmetric growth

The role of ubiquitination and deubiquitination in the regulation of cell junctions

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Protein & Cell 2018年第9期9卷 754-769页

作者： Junting Cai Miranda K. Culley Yutong Zhao Jing Zhao Acute Lung Injury Center of Excellence Division of Pulmonary Asthma and Critical Care Medicine Department of Medicine The University of Pittsburgh Pittsburgh PA 15213 USA Xiangya Hospital of Central South University Changsha 410008 China

Maintenance of cell junctions plays a crucial role in the regulation of cellular functions including cell proliferation, permeability, and cell death. Disruption of cell junctions is implicated in a variety of human disorders, such as inflammatory diseases and cancers. Understanding molecular regulation of cell junctions is important for development of therapeutic strategies for intervention of human diseases. Ubiquitination is an important type of post-translational modification that primarily regulates endogenous protein stability, recep- tor internalization, enzyme activity, and protein-protein interactions. Ubiquitination is tightly regulated by ubiq- uitin E3 ligases and can be reversed by deubiquitinating enzymes. Recent studies have been focusing on inves- tigating the effect of protein stability in the regulation of cell-cell junctions. Ubiquitination and degradation of cadherins, claudins, and their interacting proteins are implicated in epithelial and endothelial barrier disruption. Recent studies have revealed that ubiquitination is involved in regulation of Rho GTPases＇ biological activities. Taken together these studies, ubiquitination plays a critical role in modulating cell junctions and motility. In this review, we will discuss the effects of ubiquitination and deubiquitination on protein stability and expression of key proteins in the cell-cell junctions, including junction proteins, their interacting proteins, and small Rho GTPases. We provide an overview of protein stability in modulation of epithelial and endothelial barrier integrity and introduce potential future search directions to better understand the effects of ubiquitination on human disorders caused by dysfunction of cell junctions.

关键词： cell-cell junctions protein stability ubiquitination deubiquitination Rho GTPases

Signaling network controlling ROP-mediated tip growth in Arabidopsis and beyond

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Plant Communications 2023年第1期4卷 198-208页

作者： En Li Yu-Ling Zhang Zheng Qin Meng Xu Qian Qiao Sha Li Shan-Wei Li Yan Zhang State Key Laboratory of Crop Biology College of Life SciencesShandong Agricultural UniversityTai’anShandong 271018China College of Life Sciences Nankai UniversityTianjin 300071China

Cell polarity operates across a broad range of spatial and temporal scales and is essential for specific biological functions of polarized cells.Tip growth is a special type of polarization in which a single and unique polarization site is established and maintained,as for the growth of root hairs and pollen tubes in plants.Extensive studies in past decades have demonstrated that the spatiotemporal localization and activity of rho of Plants(ROPs),the only class of Rho GTPases in plants,are critical for tip growth.ROPs are switched on or off by different factors to initiate dynamic intracellular activities,leading to tip growth.Recent studies have also uncovered several feedback modules for ROP signaling.In this review,we summarize recent progress on ROP signaling in tip growth,focusing on molecular mechanisms that underlie the dynamic distribution and activity of ROPs in Arabidopsis.We also highlight feedback modules that control ROPmediated tip growth and provide a perspective for building a complex ROP signaling network.Finally,we provide an evolutionary perspective for ROP-mediated tip growth in Physcomitrella patens and during plant–rhizobia interaction.

关键词： Rho GTPases feedback polarization root hair pollen tube

“大黄-桃仁”药对调控子宫腺肌病rho GTPase的作用研究

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“大黄-桃仁”药对调控子宫腺肌病Rho GTPase的作用研究

作者：雷娅三峡大学

学位级别：硕士

研究背景:大黄桃仁药对(Dahuang-Taoren,DT)来源于中医经典方,作为活血化瘀类的经典植物药物组合,DT常用于缓解子宫腺肌症(adenomyosis,AM)的痛经症状。越来越多的证据表明Rho GTPases可能在AM进展中发挥重要作用,然而,DT对AM中rho GTPa... 详细信息

研究背景:大黄桃仁药对(Dahuang-Taoren,DT)来源于中医经典方,作为活血化瘀类的经典植物药物组合,DT常用于缓解子宫腺肌症(adenomyosis,AM)的痛经症状。越来越多的证据表明Rho GTPases可能在AM进展中发挥重要作用,然而,DT对AM中Rho GTPases的潜在调控作用尚不清楚。为此,我们对DT对AM Rho GTPases的干预作用进行了评估。目的:探讨DT对AM rho GTPase的调控作用。方法:(1)收集临床标本,对比AM患者及对照组子宫内膜组织中的RAS同源基因家族成员A(Ras homolog gene family,rho A)、rho效应激酶(rho kinase 1,Rock1)、Ras相关的C3肉毒素底物1(Ras-related C3 botulinum toxin substrate 1,Rac1)以及细胞分裂周期蛋白42(cell division cycle 42,Cdc42)蛋白水平。提取分离培养原代子宫内膜间质细胞(adenomyosis endometrial stromal cells,AMESCs),通过MTT增殖实验和划痕试验观察细胞的生存活力和迁移能力,Western blot实验检测rho A、Rock1、Rac1和Cdc42蛋白表达情况。(2)将30只ICR雌性小鼠随机分成对照组(n=10)、模型组(n=10)和DT组(n=10),AM小鼠模型采用同种异体垂体移植法建立。热板痛和机械痛实验检测小鼠痛觉敏感性,HE染色法检测子宫组织病理浸润程度,透射电镜观察细胞动力学特征变化,免疫荧光染色法检测蛋白rho A、Rock1、Rac1和Cdc42的表达水平和定位情况。(3)采用分子对接技术验证rho GTPase蛋白靶点与药物核心活性成分的结合能力。结果:(1)临床组织Western Blot实验结果显示,与对照组比较,AM患者rho A、Rac1、Cdc42、Rock1蛋白水平均明显上调。体外成功提取分离出原代AMESCs。MTT实验和划痕实验结果显示,DT剂量依赖性抑制AMESCs增殖和迁移。Western Blot实验结果显示,0.25 mg/ml DT和0.5 mg/ml DT显著抑制了AMESCs的rho A、Rock1、Cdc42和Rac1的蛋白水平。(2)动物实验结果显示,与模型组比较,经DT治疗后小鼠热板痛及机械痛的痛阈增高,病理浸润程度减轻。电镜结果表明,与对照组比较,模型组小鼠子宫内膜上皮细胞排列紊乱、体积缩小,细胞形态由柱状变为梭形或不规则形;细胞间紧密连接、桥粒连接及细胞周边指状突起减少,细胞膜局部形成皱褶。经DT治疗后上述动力学特征变化改善。免疫荧光染色结果显示,rho A、Rock1、Rac1、Cdc42可定位表达于子宫内膜腺上皮细胞、间质细胞及平滑肌细胞。与对照组比较,模型组小鼠rho A、Rock1和Rac1的表达显著上调,经DT治疗后回调。(3)分子对接结果显示,DT中的药物的活性成分与rho A、Rock1、Rac1和Cdc42均具有良好或者较强的结合能力。结论:本研究表明AM中存在Rho GTPases的过度激活。DT可有效缓解AM痛觉过敏反应、改善病理浸润程度,其作用可能与抑制Rho GTPases调控细胞动力学特征有关。

关键词：大黄-桃仁药对子宫腺肌病 Rho GTPases 分子对接

Rac1及Cdc42在肾上腺髓质肽(AM)缓解嘌呤霉素氨基核苷(PAN)所诱导足细胞损伤中作用机制的初步探讨

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复旦学报（医学版） 2020年第4期47卷 496-505页

作者：覃乔静常凯利孟利霞董楠赵仲华刘学光复旦大学附属上海市第五人民医院肾内科上海200240 复旦大学基础医学院病理学系上海200032 上海市肾脏疾病与血液净化重点实验室上海200032

目的观察肾上腺髓质肽(adrenomedullin,AM)对嘌呤霉素氨基核苷(puromycin aminonucleoside,PAN)所诱导受损足细胞的作用以及Rac1/Cdc42表达变化,初步探讨其可能机制。方法将小鼠肾足细胞株分为对照组、PAN处理组(100μg/mL)、PAN联合AM... 详细信息

目的观察肾上腺髓质肽(adrenomedullin,AM)对嘌呤霉素氨基核苷(puromycin aminonucleoside,PAN)所诱导受损足细胞的作用以及Rac1/Cdc42表达变化,初步探讨其可能机制。方法将小鼠肾足细胞株分为对照组、PAN处理组(100μg/mL)、PAN联合AM处理组(10-7mol/L)及PAN联合AM和蛋白激酶A(PKA)抑制剂H89(10-6mol/L)处理组。经一次性腹腔注射PAN(150 mg/kg)建立大鼠足细胞损伤模型,每天经尾静脉注射AM蛋白(66μg/kg)进行干预。SDS-PAGE法检测尿蛋白水平,电镜观察足细胞足突宽度变化,双重免疫荧光染色及/或Western blot观察足细胞特异性骨架蛋白(synaptopodin、nephrin)、Rac1及Cdc42的蛋白表达水平,实时定量PCR(qRT-PCR)检测synaptopodin、nephrin的mRNA表达,谷胱甘肽巯基转移酶-拉下实验(GST-pull down assay)法检测Rac1及Cdc42活性变化。结果PAN在体内、外均显著降低synaptopodin、nephrin的蛋白或/及mRNA表达水平,且显著升高desmin表达;大鼠PAN肾病模型的尿蛋白水平显著升高,足突宽度显著增加;PAN显著降低Rac1、Cdc42总蛋白及活性水平。上述由PAN所介导的效应被AM显著抑制;H89显著阻断AM的体外作用。结论AM主要通过PKA通路促进受损足细胞的修复,该保护机制与AM调控Rac1/Cdc42表达密切相关。

关键词：肾上腺髓质肽(AM) Rho GTPases 足细胞嘌呤霉素氨基核苷(PAN) 细胞骨架蛋白大鼠

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条件恐惧记忆的分子机制研究进展

生命科学 2021年第7期33卷 791-800页

作者：杨飞韩笑刘华东西安交通大学生线粒体生物医学研究所西安710049

恐惧位于记忆和情感的交汇处,既是触发防御机制以适应危险的一种情绪反应,又是研究记忆的模型。恐惧调节障碍时常会导致焦虑。该文综述了条件恐惧记忆形成及维持的主要机制,总结了在恐惧记忆产生后,神经元响应钙离子内流时相关的分子生... 详细信息

恐惧位于记忆和情感的交汇处,既是触发防御机制以适应危险的一种情绪反应,又是研究记忆的模型。恐惧调节障碍时常会导致焦虑。该文综述了条件恐惧记忆形成及维持的主要机制,总结了在恐惧记忆产生后,神经元响应钙离子内流时相关的分子生物学变化,如CaMKs、PKA、PKM-ζ、Rho GTPases等激酶的激活,以及下游MAPK信号的转导、转录因子的调控、相关基因的表达等方面的现有研究结果,试图理清恐惧记忆相关分子机制。鉴于恐惧调节与诸如创伤后应激障碍(PTSD)之类的某些情绪障碍有密切关系,条件恐惧记忆形成的分子机制研究可能为精神障碍药物的开发提供潜在的靶标。

关键词：条件恐惧记忆长时程增强 CaMKs PKA PKM-ζ CREB Rho GTPases

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