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A Complete Sojourn of Monoclonal Antibodies: AI, Rare Diseases / Disorders And Immunotoxic Effects 单克隆抗体大全》:人工智能、罕见疾病/障碍和免疫毒性效应
IF 0.8 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-30 DOI: 10.2174/0115701646313765240610062419
Sonakshi Garg, Gurisha Garg, Preeti Patel, Ghanshyam Das Gupta, Balak Das Kurmi
: Monoclonal antibodies (mAbs) are magic bullets proved to be a wonder in the pharmaceutical as well as medical fields. These are produced by various methods like hybridoma technology, phage display technology, YAC technology, and transgenic animals and plants. Based on the percentage of animal origin, mAbs are divided into chimeric, murine, humanized, and fully human. This review covers the history and methods of mAb production, immunotoxicity (Immunosuppression, immunostimulant, autoimmunity, hypersensitivity) associated with mAbs, and targets of mAbs. It also compiles mAb production using AI, new modifications, and novel mAbs, with its various clinical trial information ensuring the use of mAbs in rare diseases and disorders.
:单克隆抗体(mAbs)是制药和医疗领域的神奇子弹。它们是通过杂交瘤技术、噬菌体展示技术、YAC 技术以及转基因动物和植物等各种方法生产出来的。根据动物来源的比例,mAbs 可分为嵌合型、鼠型、人源化和全人型。本综述涵盖 mAb 生产的历史和方法、与 mAb 相关的免疫毒性(免疫抑制、免疫刺激、自身免疫、超敏反应)以及 mAb 的靶点。它还汇编了使用人工智能生产的 mAb、新的修饰和新型 mAb,以及确保 mAb 用于罕见疾病和失调症的各种临床试验信息。
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引用次数: 0
StemRegenin-1 Reverses Drug Resistance of MCF-7/ADR Cells via AhR/ABC Transports and AhR/UGTs Pathways StemRegenin-1 通过 AhR/ABC 转运和 AhR/UGTs 途径逆转 MCF-7/ADR 细胞的耐药性
IF 0.8 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-22 DOI: 10.2174/0115701646317215240712103448
Yang Zhang, Yu-Chen Ma, Jue Song, Yong Jin, Yan-Ni Bao
Objectives: Drug resistance reduces the antitumor efficacy of chemotherapy. Therefore, it is important to know how to reverse drug resistance. In this work, we investigated drug resistance reversal by StemRegenin-1(SR-1) in MCF-7/ADR cells and the mechanism by which it exerts its drug resistance effect. Methods: MTT test and protein blot were employed as the two main in vitro cell tests. The cells were treated with SR-1 and ADM to detect the changes in their proteomics, and then the effects of AhR downstream proteins, glucuronidase, and drug-resistant proteins were verified. The accumulation of ADM in the combined cells and its effect on the cell cycle were detected by flow cytometry. In vivo, a BALB/C mice xenograft test was conducted to observe the anti-tumor effect and side effects of the drug combination. Results: SR-1 combined with ADM inhibited cell proliferation and significantly decreased the expression of CYP1A1, UGT1A6, P-gP (ABCB1), and MRP1 (ABCC1). Furthermore, SR-1 caused apoptosis and cell cycle arrest. In vivo experiments showed that SR-1 significantly enhanced the antitumor effects of ADM and reduced the toxic effects of ADM. Conclusion: SR-1 inhibited AhR activity, decreased its downstream protein CYP1A1 and the expression of UGT1A6, P-gP, and MRP1 in MCF-7/ADR cells, and reversed drug resistance in MCF-7/ADR cells through AhR/ABC transports and AhR/UGTs pathways.
目的:耐药性会降低化疗的抗肿瘤疗效。因此,了解如何逆转耐药性非常重要。本研究探讨了StemRegenin-1(SR-1)在MCF-7/ADR细胞中的抗药性逆转作用及其发挥抗药性作用的机制。研究方法体外细胞检测主要采用 MTT 试验和蛋白印迹法。用 SR-1 和 ADM 处理细胞,检测其蛋白质组学的变化,然后验证 AhR 下游蛋白、葡糖醛酸酶和耐药蛋白的作用。流式细胞术检测了 ADM 在合并细胞中的积累及其对细胞周期的影响。在体内,进行了 BALB/C 小鼠异种移植试验,观察联合用药的抗肿瘤效果和副作用。结果显示SR-1 与 ADM 联用可抑制细胞增殖,并显著降低 CYP1A1、UGT1A6、P-gP (ABCB1) 和 MRP1 (ABCC1) 的表达。此外,SR-1 还能导致细胞凋亡和细胞周期停滞。体内实验表明,SR-1 能显著增强 ADM 的抗肿瘤作用,并降低 ADM 的毒性作用。结论SR-1可抑制AhR活性,降低其下游蛋白CYP1A1以及UGT1A6、P-gP和MRP1在MCF-7/ADR细胞中的表达,并通过AhR/ABC转运和AhR/UGTs途径逆转MCF-7/ADR细胞的耐药性。
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引用次数: 0
In silico Investigation on the Structural Insights into the Binding of Squalamine Inhibitor with Membrane-Bound Α-Synuclein 角鲨胺抑制剂与膜结合型Α-突触核蛋白结合的结构洞察的硅学研究
IF 0.8 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-10 DOI: 10.2174/0115701646301714240703100842
Dorothy Das, Priyam Bharadwaz, Venkata Satish Kumar Mattaparthi
Background: Parkinson's disease (PD) and its associated symptoms are closely associated with the self-assembly of α-Synuclein (α-Syn). Squalamine is a naturally occurring chemical substance with established antiviral and anticancer properties, and its profound impact on the α- Syn aggregation both in vivo and in vitro is well studied. Examining its interaction with lipid vesicles, which are known to encourage nucleation, can signify the mechanism of action of squalamine. The squalamine molecule is believed to displace α-Syn from the surfaces of the lipid vesicles, therefore preventing the initial steps in the process of aggregation. Additionally, the squalamine molecule reduces the harmful effects of α-Syn oligomers in human neuroblastoma cells by preventing them from interacting with lipid membranes. Objective: The aim of this study was to perform computational investigation of the conformational changes of membrane-bound α-Syn in the presence of squalamine inhibitor molecule objective: Computational investigation of the conformational changes of membrane-bound α-Synuclein in the presence of squalamine inhibitor molecule Method: Molecular Dynamics (MD) trajectory analysis was carried out to study the structural change of the α-Syn-squalamine conformers as a function of simulation time. The percentage of the secondary structural components of the α-Syn-squalamine complex was determined. Optimization of small molecule inhibitors was carried out using Density Functional Theory (DFT) analysis. Additionally, the values of electrophilicity (ω), nucleophilicity (N), Electron affinity (EA), and ionization potential (IP) were calculated. Results: The docking of the α-Syn-squalamine complex revealed the binding site and the best structure was selected based on the highest docking vina score (-5.8), and the contact residues were listed. From the conformational snapshots of the α-Syn-squalamine complex, it was evident that the α-Syn remained stable, maintaining its integrity throughout the simulation. The α-helical content was found to be retained from the secondary structural content analysis. The ω and N of the squalamine molecule were calculated to be -0.84 and 3.25, respectively. Conclusion: Our findings suggest that in the presence of a squalamine inhibitor molecule, α-Syn adopts a helical conformation that ensures stability and may indicate that the squalamine molecule causes gradual displacement of α-Syn across different regions within the lipid membrane.
背景:帕金森病(PD)及其相关症状与α-突触核蛋白(α-Syn)的自组装密切相关。角鲨胺是一种天然化学物质,具有公认的抗病毒和抗癌特性,其对体内和体外α-突触核蛋白聚集的深远影响已得到充分研究。研究它与脂质囊泡的相互作用(众所周知,脂质囊泡会促进成核)可以揭示角鲨胺的作用机制。据信,角鲨胺分子会将α-Syn从脂质囊泡表面置换出来,从而阻止聚集过程的初始步骤。此外,角鲨胺分子还能阻止α-Syn寡聚体与脂质膜相互作用,从而减少α-Syn寡聚体对人类神经母细胞瘤细胞的有害影响。研究目的本研究的目的是对角鲨胺抑制剂分子存在时膜结合的α-Syn的构象变化进行计算研究:计算研究膜结合的α-突触核蛋白在角鲨胺抑制剂分子存在下的构象变化:通过分子动力学(MD)轨迹分析,研究α-Syn-角鲨胺构象的结构变化与模拟时间的函数关系。确定了α-Syn-喹啉胺复合物二级结构成分的百分比。利用密度泛函理论(DFT)分析对小分子抑制剂进行了优化。此外,还计算了亲电性(ω)、亲核性(N)、电子亲和力(EA)和电离电位(IP)的值。结果表明对α-Syn-草酰氨基复合物的对接揭示了其结合位点,并根据最高的对接 vina 得分(-5.8)选出了最佳结构,列出了接触残基。从α-Syn-水杨酸胺复合物的构象快照可以看出,α-Syn保持稳定,在整个模拟过程中都保持其完整性。二级结构成分分析发现,α-螺旋成分得以保留。计算得出角鲨胺分子的ω和N分别为-0.84和3.25。结论我们的研究结果表明,在角鲨胺抑制剂分子存在的情况下,α-Syn会采用螺旋构象以确保其稳定性,这可能表明角鲨胺分子会导致α-Syn在脂膜内的不同区域逐渐移位。
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引用次数: 0
Comparison of the Effects of Different Coating Matrices on Cell Binding and Nitric Oxide-Mediated Protein S-Nitrosylation in Endothelial Cellsunder Shear Flow 比较不同涂层基质对剪切流下内皮细胞中细胞结合和一氧化氮介导的蛋白 S-亚硝基化的影响
IF 0.8 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-06-13 DOI: 10.2174/0115701646300960240606093535
Ming-Chung Lin, Ming-Wei Lin, Erna Sulistyowati, Ching-Chieh Kao, Chung-Jung Liu, Shu-Ping Huang, S. C. Hsu, Bin Huang
Shear flow is a mechanical signal regulating the function of EndothelialCells (ECs). The present study aimed to investigate the effects of different matrices on cell binding,Nitric Oxide (NO) production, protein S-nitrosylation, expression of adhesion proteins, ROSgeneration, and cell viability in ECs under shear flow.The ECs growing on glass slides separately coated with poly-L-lysine (p-Lys), collagen(Colla), fibronectin (Fibro), and a combined matrix (Colla+Fibro) were exposed to shear flow (25dyne/cm2) for 0, 1, 4, 8 h. The number of ECs remaining attached on the glass slide was calculated.The expressions of endothelial Nitric Oxide Synthase (eNOS), peNOSS1177, VE-cadherin, FAK,and S-nitrosylated proteins were investigated by western blotting. The production of Nitric Oxide(NO) was measured by a specific reagent. Finally, the levels of ROS and cell viability were monitored.Under a constant shear flow for 1 h, the physiological responses of ECs were similar betweenthese four matrices. When shear flow was extended to 4 and 8 h, higher cell binding, elevatedNO production, increased S-nitrosylated proteins, enhanced expressions of FAK and VE-cadherin,mildly accumulated ROS, and cell death were observed in the matrix of Fibro and Colla+Fibro.We have concluded fibronectin to be the optimal matrix facilitating NO-mediated Snitrosylationthat might be essential for superior binding efficiency, thereby preventing the strippingof ECs under shear flow. The results can be broadly applied to diverse biomechanicalstudies.
剪切流是一种调节内皮细胞(EC)功能的机械信号。本研究旨在探讨不同基质对剪切流下内皮细胞的细胞结合、一氧化氮(NO)产生、蛋白质 S-亚硝基化、粘附蛋白表达、ROS 生成和细胞活力的影响。将分别涂有聚-L-赖氨酸(p-Lys)、胶原(Colla)、纤连蛋白(Fibro)和组合基质(Colla+Fibro)的玻璃载玻片上生长的心肌细胞暴露于剪切流(25dne/cm2)中 0、1、4、8 小时。用 Western 印迹法检测内皮一氧化氮合酶(eNOS)、peNOSS1177、VE-cadherin、FAK 和 S-亚硝基化蛋白的表达。一氧化氮(NO)的产生是通过特定试剂测定的。在恒定剪切流 1 小时的条件下,四种基质对 EC 的生理反应相似。当剪切流延长到4小时和8小时时,在纤维和阿胶+纤维基质中观察到更高的细胞结合率、NO产生量增加、S-亚硝基化蛋白增加、FAK和VE-cadherin表达增强、ROS轻度积累以及细胞死亡。这些结果可广泛应用于各种生物力学研究。
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引用次数: 0
An Immunoinformatic Approach to Designing a Multi-epitope Vaccine against Helicobacter pylori with the VacA Toxin and BabA Adhesion 利用 VacA 毒素和 BabA 吸附设计幽门螺旋杆菌多表位疫苗的免疫形式化方法
IF 0.8 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-05-31 DOI: 10.2174/0115701646302487240524103934
Viana Dayhimi, Fatemeh Ziadlou, Simin Nafian, Fatemeh Nafian
Objective: Helicobacter pylori, as a carcinogen, has been related to the development of gastric cancer, particularly in developing countries. The main challenge with therapy is the recurrence of antibiotic-resistant bacteria, and vaccination is still a problem. Therefore, the objective of the current study was to rationally design a multi-epitope vaccine using two immunogenic proteins found in H. pylori. Methods: Promising epitopes for the Leb-binding adhesin A (BabA) and vacuolating cytotoxin (VacA) proteins were characterized through an immunoinformatics approach. Epitope-rich fragments were selected based on high-binding affinities with HLA classes I and II to be specifically presented to B and T lymphocytes and to selectively elicit both humoral and cellular immune responses. Results: Six constructs were planned by fusing these fragments in different arrangements with the help of GPGPG linkers. The most stable three-dimensional structure was found in Construct 6 during molecular dynamics. To improve immunogenicity and stability, an adjuvant called human β- defensin 2 (hBD-2) was joined to the N-terminus of Construct 6. Following molecular docking, the final vaccine reacted appropriately with each toll-like receptor 2 (TLR-2), TLR3, and TLR-4. The final DNA sequence was optimized for expression in E. coli K12 and in silico cloned into a pET-28a(+) plasmid. As a result of the vaccination in silico, substantial responses were developed against H. pylori. Conclusion: According to the immune response simulation, activated B and T lymphocytes and memory cell production increased. Macrophages and dendritic cells proliferated continuously, and IFN-γ and Cytokines, such as IL-2 were raised.
目的:幽门螺杆菌是一种致癌物质,与胃癌的发病有关,尤其是在发展中国家。治疗的主要挑战是耐抗生素细菌的复发,而疫苗接种仍是一个问题。因此,本研究的目的是利用幽门螺杆菌中的两种免疫原蛋白合理设计一种多表位疫苗。方法通过免疫信息学方法确定了Leb结合粘附素A(BabA)和空泡细胞毒素(VacA)蛋白的有望表位。根据与 HLA I 类和 II 类的高结合亲和力筛选出表位丰富的片段,以便特异性地呈现给 B 淋巴细胞和 T 淋巴细胞,并选择性地引起体液免疫和细胞免疫反应。结果:在 GPGPG 连接器的帮助下,将这些片段以不同的排列方式融合在一起,规划出了六种构建体。在分子动力学中,构建体 6 的三维结构最为稳定。为了提高免疫原性和稳定性,在构建体 6 的 N 端加入了一种名为人 β 防御素 2(hBD-2)的佐剂。经过分子对接,最终的疫苗与收费样受体 2(TLR-2)、TLR3 和 TLR-4 均有适当的反应。最终的 DNA 序列经过优化,可在大肠杆菌 K12 中表达,并被克隆到 pET-28a(+)质粒中。由于在硅学中进行了疫苗接种,对幽门螺杆菌产生了实质性的反应。结论根据免疫反应模拟,活化的 B 淋巴细胞和 T 淋巴细胞以及记忆细胞的生成都有所增加。巨噬细胞和树突状细胞不断增殖,IFN-γ 和细胞因子(如 IL-2)升高。
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引用次数: 0
Investigating PI3P Binding with Plasmodium Falciparum HSP70 Proteins 研究 PI3P 与恶性疟原虫 HSP70 蛋白的结合
IF 0.8 4区 生物学 Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-04-17 DOI: 10.2174/0115701646297476240408042556
Vipul Upadhyay, Satinder Kaur, Rachna Hora, Prakash Chandra Mishra
aims: Investigating the binding of phosphatidylinositol 3 phosphate (PI3P) with Plasmodium falciparum heat shock protein 70 homologs background: The 70 kDa Plasmodium falciparum (Pf) heat shock proteins (PfHSP70s) are an important class of molecules that are critically involved in parasite survival through periods of stress. Interaction between a crucial lipid modulatorPI3P and the C terminal lid domain of cytosolic PfHSP70-1, stabilizes the parasite digestive vacuole (DV) to facilitate hemoglobin trafficking and breakdown in turn impacting parasite survival. PfHSP70 homologs and PI3P are amply expressed together in various subcellular compartments of the parasite providing these with an opportunity to interact and affect biological processes. objective: The objectives of this study included identification of the PI3P binding pockets on PfHSP70s, understanding the binding affinity of each homolog for PI3P and identifying residues involved in different types of interactions. method: We have analyzed PI3P binding of all four PfHSP70s by using bioinformatics tools like sequence analysis, molecular docking and LigPlot+ analysis. result: Our results show that differently localized PfHSP70 homologs bind PI3P with variable affinity. The mitochondrial and ER localized PfHSP70s bind more strongly with PI3P than their cytosolic counterpart PfHSP70-1. The PI3P binding region on all PfHSP70 homologs lies at the interface of helices 1 and 3 of their substrate binding domain . Analysis of these results has also helped to pinpoint specific residues on PfHSP70s that may be engaged in these interactions. conclusion: PI3P preferentially binds to a particular pocket on PfHSP70 homologs. Different PfHSP70s bind to PI3P with variable binding affinity. Specific residues of PfHSP70 homologs involved in these interactions have been identified. other: The present study may therefore form the basis for designing interventions that hinder PfHSP70-PI3P interaction and influence parasite survival.
目的研究磷脂酰肌醇 3 磷酸酯(PI3P)与恶性疟原虫热休克蛋白 70 同源物的结合背景:70 kDa 的恶性疟原虫(Pf)热休克蛋白(PfHSP70s)是一类重要的分子,对寄生虫在应激期的生存起着至关重要的作用。一种重要的脂质调节剂PI3P与细胞质PfHSP70-1的C端盖状结构域相互作用,稳定了寄生虫消化泡(DV),促进了血红蛋白的运输和分解,进而影响寄生虫的存活。PfHSP70 同源物和 PI3P 在寄生虫的各种亚细胞区室中大量表达,为它们提供了相互作用和影响生物过程的机会:本研究的目的包括鉴定 PfHSP70s 上的 PI3P 结合口袋,了解每个同源物与 PI3P 的结合亲和力,以及鉴定参与不同类型相互作用的残基:我们利用序列分析、分子对接和 LigPlot+ 分析等生物信息学工具分析了所有四种 PfHSP70s 的 PI3P 结合情况:我们的结果表明,不同定位的 PfHSP70 同源物以不同的亲和力结合 PI3P。线粒体和 ER 定位的 PfHSP70 与 PI3P 的结合比其细胞质对应物 PfHSP70-1 更强。所有 PfHSP70 同源物上的 PI3P 结合区都位于其底物结合域  的第 1 和第 3 螺旋的界面上。对这些结果的分析还有助于确定 PfHSP70 上可能参与这些相互作用的特定残基:PI3P 优先与 PfHSP70 同源物上的一个特定口袋结合。不同的 PfHSP70 与 PI3P 的结合亲和力不同。参与这些相互作用的 PfHSP70 同源物的特定残基已被确定:因此,本研究可为设计阻碍 PfHSP70-PI3P 相互作用并影响寄生虫存活的干预措施奠定基础。
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引用次数: 0
Designing a Novel Multi-Epitope Peptide as a Potential Serodiagnosis Marker for the Diagnosis of Acinetobacter baumannii: An In silico Approach 设计一种新型多表位肽作为诊断鲍曼不动杆菌的潜在血清诊断标记物:硅学方法
IF 0.8 4区 生物学 Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-04-02 DOI: 10.2174/0115701646297689240325062145
Maryam Rezaee, Mohsen Mohammadi, Amir Savardashtaki, Mohammad Reza Rahbar, Navid Nezafat
Background: Acinetobacter baumannii is an opportunistic pathogen that causes many infections, including nosocomial infections; this bacterium has a high mortality rate among other bacteria. A. baumannii has an elastic genome that changes rapidly when exposed to harsh environmental conditions, leading to widespread bacterial resistance to various disinfectants and antibiotics. The high ability of bacteria to bind to all surfaces and survive in different conditions has caused the spread of bacteria in various environments. Rapid detection is very important in preventing the spread and even treatment of the infection. Methods: Currently, the Polymerase Chain Reaction (PCR) method is the only effective method used for diagnosis, which has some pros and cons. Results and Conclusion: This study aimed to design a new recombinant multi-epitope protein from Acinetobacter baumannii that can be used in ELISA for rapid diagnosis. The unique feature of this study from others is the use of patient serum for antibody monitoring.
背景:鲍曼不动杆菌(Acinetobacter baumannii)是一种机会性病原体,可引起多种感染,包括医院内感染;在其他细菌中,这种细菌的死亡率很高。鲍曼不动杆菌的基因组具有弹性,当暴露在恶劣的环境条件下时,基因组会迅速发生变化,从而导致细菌对各种消毒剂和抗生素产生广泛的抗药性。细菌具有很强的与各种表面结合的能力,并能在不同的条件下存活,这就造成了细菌在各种环境中的传播。快速检测对于防止感染扩散甚至治疗都非常重要。方法:目前,聚合酶链反应(PCR)法是唯一有效的诊断方法,但这种方法有利有弊。结果与结论:本研究旨在设计一种新的重组鲍曼不动杆菌多表位蛋白,可用于 ELISA 快速诊断。与其他研究相比,本研究的独特之处在于使用患者血清进行抗体监测。
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引用次数: 0
Proteomic Analysis of the Molecular Mechanisms of Chlorpromazine Inhibiting Migration of Oral Squamous Cell Carcinoma 氯丙嗪抑制口腔鳞状细胞癌迁移的分子机制蛋白质组学分析
IF 0.8 4区 生物学 Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-02-27 DOI: 10.2174/0115701646291510240212091951
Nannan Zhang, Junzhi Liu, Qiuping Dong, Chen Liu, Xinyu Liang, Peiyuan Tang, Zheng Liang
Background: Oral squamous cell carcinoma (OSCC) is a common malignant tumor of the head and neck region known for its high metastatic and invasive potential. Chlorpromazine (CPZ) has been shown to inhibit the growth of oral cancer cells. However, the effects of CPZ on OSCC migration and its underlying molecular mechanisms remain unclear. Objective: We aimed to identify global protein changes and potential core proteins involved in CPZ-mediated inhibition of migration in SCC-15 cells using proteomics. Methods: We assessed the effect of CPZ on SCC-15 using CCK-8 assays and wound healing experiments. Next, we performed LC-MS-based proteomic analysis to identify protein alterations in SCC-15 cells treated with CPZ at different times. Differential expression proteins (DEPs) were identified and subjected to bioinformatics analysis using GO, KEGG, and PPI tools. Key candidate proteins were selected and validated using the TCGA-HNSCC database and molecular docking. Results: It was found that 20μm of CPZ had no effect on cell proliferation, but inhibited cell migration. A total of 4748 proteins were identified by Proteomics, among which 56 DEPs were identified, including 34 upregulated proteins and 22 downregulated proteins. Three proteins (RPF2, ACTB, and TGFBI) were identified as key candidate proteins associated with cell adhesion and migration in oral cancer cells. Conclusion: CPZ may affect the expression of RPF2, ACTB, and TGFBI proteins and change the extracellular matrix and cell adhesion function, thus inhibiting the migration of SCC-15 cells. The results of this study provide a robust basis for further research on the molecular mechanism of CPZ to inhibit the migration of OSCC.
背景:口腔鳞状细胞癌(OSCC)是头颈部常见的恶性肿瘤,具有高转移性和侵袭性。氯丙嗪(CPZ)已被证明能抑制口腔癌细胞的生长。然而,CPZ 对 OSCC 迁移的影响及其潜在的分子机制仍不清楚。研究目的我们的目的是利用蛋白质组学鉴定 CPZ 介导的 SCC-15 细胞迁移抑制所涉及的全局蛋白变化和潜在核心蛋白。方法: 我们评估了 CPZ 对 SCC-15 细胞迁移的影响:我们使用 CCK-8 试验和伤口愈合实验评估了 CPZ 对 SCC-15 的影响。接着,我们进行了基于 LC-MS 的蛋白质组学分析,以确定不同时间用 CPZ 处理的 SCC-15 细胞中的蛋白质变化。我们利用 GO、KEGG 和 PPI 工具鉴定了差异表达蛋白(DEPs)并对其进行了生物信息学分析。利用 TCGA-HNSCC 数据库和分子对接技术筛选并验证了关键候选蛋白。结果发现结果发现,20μm 的 CPZ 对细胞增殖没有影响,但抑制了细胞迁移。蛋白质组学共鉴定出4748个蛋白质,其中56个DEPs,包括34个上调蛋白质和22个下调蛋白质。三个蛋白质(RPF2、ACTB和TGFBI)被确定为与口腔癌细胞粘附和迁移相关的关键候选蛋白质。结论CPZ 可能会影响 RPF2、ACTB 和 TGFBI 蛋白的表达,改变细胞外基质和细胞粘附功能,从而抑制 SCC-15 细胞的迁移。本研究结果为进一步研究 CPZ 抑制 OSCC 迁移的分子机制提供了有力依据。
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引用次数: 0
Immunoprotective Potential of Adenylosuccinate Synthetase Protein (PurA) in Streptococcus equi ssp. zooepidemicus Infections 腺苷酸合成酶蛋白(PurA)在马链球菌感染中的免疫保护潜力
IF 0.8 4区 生物学 Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-02-23 DOI: 10.2174/0115701646284439240218063821
Xiaoling Zhang, Sihuan Chen, Shuji Gao, Weiping Yang, Yuxin Wang, Yang Wang, Li Yi
Background: Streptococcus equi ssp. zooepidemicus(SEZ) is one important pathogen. There are still sporadic outbreaks in China, northern United States and the Netherlands. Adenylosuccinate synthetase PurA, a newly discovered protein in prior research, requires further assessment of its protective effectiveness. Methods: In this study, we focused on the expression of recombinant PurA from SEZ ATCC 35246. We evaluated the immunoreactivity of this recombinant protein using convalescent minipig sera. Additionally, we conducted experiments in mice to assess its immunogenic properties. Results: Our findings revealed that the recombinant PurA triggered a substantial antibody response in mice, resulting in an 80% protection rate against SEZ infection. Notably, mice immunized with PurA exhibited significantly reduced bacterial colonization in all organs compared to the PBS control group. Furthermore, the levels of IL-6, IL-8, IL-1β, and TNF-α in mouse serum were significantly elevated in the PurA-immunized group compared to the control group. Hyperimmune sera targeting PurA effectively eliminated SEZ in bactericidal tests. Remarkably, antibodies against PurA demonstrated a significant inhibitory effect on developing SEZ biofilm. Conclusion: Immunization with PurA elicited robust humoral and cellular immune responses in mice. These promising results suggest the potential utility of PurA in developing SEZ vaccine immunogens, providing a valuable avenue for further research into SEZ infection prevention and control.
背景:马链球菌(Streptococcus equi ssp. zooepidemicus,SEZ)是一种重要的病原体。在中国、美国北部和荷兰仍有零星爆发。腺苷琥珀酸合成酶 PurA 是之前研究中新发现的一种蛋白质,需要进一步评估其保护效果。方法:在本研究中,我们重点研究了来自 SEZ ATCC 35246 的重组 PurA 的表达。我们使用痊愈小鼠血清评估了这种重组蛋白的免疫活性。此外,我们还在小鼠身上进行了实验,以评估其免疫原性。结果:我们的研究结果表明,重组 PurA 在小鼠体内引发了大量抗体反应,使小鼠对 SEZ 感染的保护率达到 80%。值得注意的是,与 PBS 对照组相比,用 PurA 免疫的小鼠在所有器官中的细菌定植率都明显降低。此外,与对照组相比,PurA 免疫组小鼠血清中的 IL-6、IL-8、IL-1β 和 TNF-α 水平明显升高。在杀菌试验中,针对 PurA 的超免疫血清能有效清除 SEZ。值得注意的是,PurA 抗体对 SEZ 生物膜的形成有明显的抑制作用。结论用 PurA 免疫可引起小鼠强有力的体液和细胞免疫反应。这些令人鼓舞的结果表明,PurA 在开发 SEZ 疫苗免疫原方面具有潜在的实用性,为 SEZ 感染预防和控制的进一步研究提供了宝贵的途径。
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引用次数: 0
Knockdown of miR-135a-5p Promotes Mitophagy by Regulating FoxO1/PINK1/Parkin Signaling in Hepatoma Cells Exposed to Oxidative Stress 敲除 miR-135a-5p 可通过调控暴露于氧化应激的肝癌细胞中的 FoxO1/PINK1/Parkin 信号促进有丝分裂
IF 0.8 4区 生物学 Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2023-12-11 DOI: 10.2174/0115701646258315231102070151
Wang Zhenchang, Zhang Wenf, Wu Shanshan, Yang Lei
Introduction: Excessive oxidative stress is always associated with hepatic disease, including hepatitis, liver fibrosis, and hepatocellular carcinoma (HCC). Despite this, the intricate molecular processes driving hepatocyte apoptosis due to oxidative stress remain incompletely comprehended. Aim: Consequently, we aimed to explore the role of miR-135a-5p in hepatoma cells (HepG2/3B). Methods: The assessment of protein expression was conducted through western blotting. Furthermore, miR-135a-5p expression was evaluated through RT-qPCR, and apoptosis detection was performed using a flow cytometry assay. Result: The findings suggest a connection between miR-135a-5p and mitochondrial-driven apoptosis through caspase signaling pathways. Furthermore, miR-135a-5p suppression inhibited the apoptotic response triggered by H2O2, reactive oxygen species (ROS) generation, as well as the decrease in mitochondrial membrane potential. Conclusion: Additionally, miR-135a-5p knockdown promoted mitophagy by regulating FoxO1/PINK1/Parkin signaling via targeting FoxO1. To conclude, our study implied that miR135a-5p might function as a probable regulator that protects cells against oxidative stress.
导言:过度的氧化应激总是与肝脏疾病相关,包括肝炎、肝纤维化和肝细胞癌(HCC)。尽管如此,人们对氧化应激导致肝细胞凋亡的复杂分子过程仍不甚了解。目的:因此,我们旨在探索 miR-135a-5p 在肝癌细胞(HepG2/3B)中的作用。方法通过 Western 印迹法评估蛋白表达。此外,还通过 RT-qPCR 评估了 miR-135a-5p 的表达,并使用流式细胞术检测了细胞凋亡。结果研究结果表明,miR-135a-5p 与线粒体通过 caspase 信号通路驱动的细胞凋亡有关。此外,抑制 miR-135a-5p 可抑制 H2O2 引发的凋亡反应、活性氧(ROS)生成以及线粒体膜电位的降低。结论此外,miR-135a-5p 的敲除还能通过靶向 FoxO1 调节 FoxO1/PINK1/Parkin 信号转导来促进有丝分裂。总之,我们的研究表明,miR135a-5p 可能是一种保护细胞免受氧化应激的调节因子。
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引用次数: 0
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Current Proteomics
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