Biochimica et biophysica acta. General subjects最新文献_第9页

Nitric oxide in plant stress: Rewilding and restoring signaling for enhancing plant growth and development 植物胁迫中的一氧化氮：重新野生和恢复促进植物生长和发育的信号

IF 2.8 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-06-20 DOI: 10.1016/j.bbagen.2025.130837

Sumreen Amin Shah , Awdhesh Kumar Mishra , Abdul Rehaman , Sumit G. Gandhi , Arif Tasleem Jan

Plants, represented as a complex system, are continuously exposed to environmental conditions that affect their growth and development, and sometimes their survival. Being sessile, they complete their life cycle under the influence of varied environmental constraints (biotic and abiotic), that adversely affect the produce's quality and productivity. Plants have evolved several defense strategies orchestrated through phytohormones that play a pivotal role in conferring resistance to stress. Nitric oxide (NO), an endogenously produced gaseous hormone, has emerged as a saviour in plant's response to different stresses. It plays an active role in the growth and development of plants, from seed dormancy and germination to growth, differentiation, flowering, fruiting, and ripening, besides affecting key metabolic processes such as photosynthesis. Endogenous production of NO and its interaction with phytohormones across different signaling cascades helps in alleviating the cellular damage caused by free radicals during drought, salinity, and other stresses. It contributes to stress resilience by inducing the synthesis of stress hormones such as ethylene (ET), which help plants to withstand adverse environmental constraints by minimizing the damage caused by different stresses. Exogenous application of NO exerts protective effects against different stresses by breaking seed dormancy and modulating germination, enhancing acquisition of mineral nutrients, photosynthetic functioning, production of antioxidant enzymes capable of neutralizing free radicals, and maintaining membrane integrity. These multifaceted roles of NO underscore its significance in plant stress tolerance. The present study offers valuable insights into NO production methods, involvement in growth and development, and a mechanistic view of its role in alleviating different stresses. In the current scenario, continued research into NO signaling mechanisms and cross-talk with other pathways seems essential for harnessing its potential in developing crops with enhanced resilience to environmental challenges.

植物作为一个复杂的系统，不断地暴露在影响其生长发育，有时甚至影响其生存的环境条件下。它们是不稳定的，在各种环境约束（生物和非生物）的影响下完成其生命周期，这对农产品的质量和生产力产生不利影响。植物已经进化出几种防御策略，这些策略是通过植物激素来协调的，这些激素在赋予植物抵抗压力的能力中起着关键作用。一氧化氮（NO）是一种内源性气体激素，在植物对不同胁迫的反应中起着救星的作用。它在植物的生长发育过程中起着积极的作用，从种子休眠、萌发到生长、分化、开花、结果、成熟，并影响光合作用等关键代谢过程。内源性NO的产生及其通过不同的信号级联与植物激素的相互作用有助于减轻自由基在干旱、盐度和其他胁迫下造成的细胞损伤。它通过诱导乙烯（ET）等应激激素的合成，帮助植物抵御不利的环境约束，最大限度地减少不同胁迫造成的损害，从而有助于抗逆性。外源施用NO通过打破种子休眠和调节萌发、增强矿质营养物质的获取、促进光合作用、产生能够中和自由基的抗氧化酶和维持膜完整性等方式对不同胁迫产生保护作用。一氧化氮在植物抗逆性中的重要作用。本研究对NO的产生方法、参与生长和发育以及其在缓解不同应激中的作用提供了有价值的见解。在目前的情况下，对NO信号机制的持续研究以及与其他途径的相互作用似乎对于利用其潜力开发具有增强环境适应能力的作物至关重要。

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引用次数: 0

USP33-mediated stabilization of c-Myc drives glycolytic reprogramming and promotes ovarian cancer progression usp33介导的c-Myc稳定驱动糖酵解重编程并促进卵巢癌进展。

IF 2.8 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-06-16 DOI: 10.1016/j.bbagen.2025.130830

Dejia Chen , Yue Zhao , Xiaobo Zhang , Xiaocheng Shi , Yiming Liu , Ge Lou

Ovarian cancer (OC) is one of the most lethal gynecological malignancies, characterized by late-stage presentation, high recurrence rates, and a lack of effective early diagnostic markers. Recent evidence suggests that deubiquitinating enzymes (DUBs) play pivotal roles in tumor development and metabolic reprogramming. Here, we identify and characterize the function of the deubiquitinase USP33 in regulating c-Myc stability and glycolytic metabolism in OC. Through quantitative PCR (qPCR) and Western blot analyses, we show that USP33 is significantly upregulated in both OC tissues and cell lines compared to normal controls. Functional assays reveal that USP33 knockdown markedly inhibits cell proliferation, migration, and invasion while promoting apoptosis. Metabolically, USP33 silencing reduces glucose uptake, lactate production, and the extracellular acidification rate, consistent with downregulation of key glycolytic enzymes (LDHA, GLUT1, and PKM2). Mechanistically, co-immunoprecipitation and ubiquitination assays demonstrate that USP33 interacts with and deubiquitinates c-Myc at K48-linked chains, thereby stabilizing c-Myc protein levels and enhancing its transcriptional activity. Moreover, c-Myc overexpression rescues the inhibitory effects of USP33 knockdown on both glycolysis and malignant phenotypes. Clinically, high USP33 expression correlates with poor prognosis, suggesting that the USP33–c-Myc axis may serve as both a prognostic biomarker and a potential therapeutic target. Taken together, our findings highlight a critical role for USP33 in OC pathogenesis by mediating c-Myc-driven glycolytic reprogramming, and they provide new insights for developing targeted treatment strategies aimed at disrupting this pathway.

卵巢癌（OC）是最致命的妇科恶性肿瘤之一，其特点是晚期出现，复发率高，缺乏有效的早期诊断标志物。最近的证据表明，去泛素化酶（DUBs）在肿瘤的发展和代谢重编程中起着关键作用。在这里，我们确定并表征了去泛素酶USP33在OC中调节c-Myc稳定性和糖酵解代谢的功能。通过定量PCR （qPCR）和Western blot分析，我们发现与正常对照相比，USP33在OC组织和细胞系中均显著上调。功能分析显示，USP33敲低显著抑制细胞增殖、迁移和侵袭，同时促进细胞凋亡。在代谢方面，USP33沉默降低了葡萄糖摄取、乳酸生成和细胞外酸化速率，这与关键糖酵解酶（LDHA、GLUT1和PKM2）的下调一致。机制上，共免疫沉淀和泛素化实验表明，USP33与k48链上的c-Myc相互作用并使其去泛素化，从而稳定c-Myc蛋白水平并增强其转录活性。此外，c-Myc过表达恢复了USP33敲低对糖酵解和恶性表型的抑制作用。临床上，USP33高表达与不良预后相关，提示USP33-c- myc轴既可作为预后生物标志物，也可作为潜在的治疗靶点。综上所述，我们的研究结果强调了USP33通过介导c- myc驱动的糖酵解重编程在OC发病机制中的关键作用，并为开发旨在破坏这一途径的靶向治疗策略提供了新的见解。

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引用次数: 0

MYO9B deficiency promoted head and neck tumor progression through HIF1α/MYC/STAT2 signaling pathway MYO9B缺乏通过HIF1α/MYC/STAT2信号通路促进头颈部肿瘤进展。

IF 2.8 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-06-13 DOI: 10.1016/j.bbagen.2025.130834

Simin Gao , Xia Gao , Ning Wang , Yuping Xie , Yuedi Tang

Background

Myosin, a protein family primarily involved in muscle contraction and cell movement, plays critical roles in various biological processes. Increasing evidence suggests its implication in tumor development and progression. However, the underlying mechanism remains poorly understood. Our study aimed to explore the role and molecular mechanism of myosin IXB (MYO9B) in head and neck squamous cell carcinoma (HNSC) development.

Methods

Utilizing The Cancer Genome Atlas (TCGA) database (n = 24) and analyzing clinical HNSC tissues (n = 24), we investigated the correlation between myosin encoding genes and HNSC progression. Cell proliferation and migration were assessed using CCK8 and Transwell assays to elucidate the tumor suppressive role of MYO9B. Additionally, Western blotting and immunostaining were conducted to evaluate the activation of HIF1α/c-Myc/STAT2 signaling. A 3D Matrigel Primary tumor cell culture model was established to assess the cisplatin resistance of HNSC cells.

Results

We found that MYO9B deficiency predicted poor prognosis in HNSC patients. In vitro, inhibiting MYO9B enhanced the proliferative and migratory characteristics of HNSC cell lines. Mechanistically, our study showed that MYO9B deficiency upregulated HIF1α signaling, leading to c-Myc upregulation, which induced stem-like phenotypes in cancer cells and HNSC progression. Furthermore, c-Myc upregulated downstream STAT2 signaling, contributing to cisplatin resistance in HNSC cells. Blocking STAT2 signaling improved cisplatin outcomes in primary MYO9B-high HNSC tissues from patients.

Conclusion

Our study highlights the tumor suppressive role of MYO9B through HI1F1α/c-Myc/STAT2 in HNSC, suggesting its potential as a diagnostic indicator and therapeutic target for clinical intervention.

背景：肌球蛋白是一个主要参与肌肉收缩和细胞运动的蛋白家族，在各种生物过程中起着关键作用。越来越多的证据表明它与肿瘤的发生和发展有关。然而，其潜在的机制仍然知之甚少。本研究旨在探讨肌球蛋白IXB （MYO9B）在头颈部鳞状细胞癌（HNSC）发生发展中的作用及分子机制。方法：利用肿瘤基因组图谱（Cancer Genome Atlas， TCGA）数据库（n = 24），分析HNSC临床组织（n = 24），探讨肌球蛋白编码基因与HNSC进展的相关性。采用CCK8和Transwell检测细胞增殖和迁移，以阐明MYO9B的抑瘤作用。此外，通过Western blotting和免疫染色来评估HIF1α/c-Myc/STAT2信号的激活情况。建立三维Matrigel原代肿瘤细胞培养模型，评估HNSC细胞对顺铂的耐药性。结果：我们发现MYO9B缺乏预示HNSC患者预后不良。在体外，抑制MYO9B可增强HNSC细胞系的增殖和迁移特性。在机制上，我们的研究表明MYO9B缺乏上调HIF1α信号，导致c-Myc上调，从而诱导癌细胞的干细胞样表型和HNSC进展。此外，c-Myc上调下游STAT2信号，促进HNSC细胞的顺铂耐药。阻断STAT2信号可改善患者原发性myo9b高HNSC组织的顺铂治疗结果。结论：本研究突出了MYO9B通过HI1F1α/c-Myc/STAT2在HNSC中的抑瘤作用，提示其可能作为临床干预的诊断指标和治疗靶点。

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引用次数: 0

The polyphenols phloretin and quercetin are potent horseradish peroxidase (HRP) inhibitors 多酚类根皮素和槲皮素是有效的辣根过氧化物酶（HRP）抑制剂。

IF 2.8 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-06-13 DOI: 10.1016/j.bbagen.2025.130833

Ben Faerman , Olivia Chalifoux , Marek Michalak , Luis B. Agellon , Ryan J. Mailloux

The discovery of horseradish peroxidase (HRP) has been highly advantageous because its unique chemistry can be applied to diagnostic tools, including the detection of oxidative distress markers like hydrogen peroxide (H₂O₂) in various experimental systems. Here, we made the surprising and compelling finding that the flavonoids, phloretin and quercetin, which are usually described in the literature as potent antioxidants, strongly inhibit the activity of HRP. Using the amplex ultrared (AUR) assay, we discovered that phloretin at a concentration as low as 50 μM abolishes the detection of H₂O₂ production by isolated liver mitochondria oxidizing pyruvate and malate. Phloretin also nullified the detection of H₂O₂ produced by liver mitochondria oxidizing succinate or dihydroorotate. Moreover, phloretin at 100 μM completely abolished the direct detection of H₂O₂ by AUR and quenched the detection of purified xanthine oxidase (XO) activity, but did not interfere with dichlorodihydrofluorescein diacetate (H₂-DCFDA) or dihydroethidine (DHE) fluorescent assays. Dose response assays revealed quercetin is a more potent inhibitor for HRP when compared to phloretin. Indeed, quercetin abolished resorufin fluorescence in AUR assays in the nM range whereas phloretin had no effect when detecting H₂O₂ in vitro or when it is formed by isolated liver mitochondria or cultured Huh-7 hepatoma and Mia-PaCa2 cells. Collectively, our findings demonstrate phloretin and quercetin, and potentially other polyphenols, potently interfere with HRP-dependent assays, which have strong implications for designing experiments that interrogate the antioxidant potential of flavonoids. Our results also indicate phloretin and quercetin could be applied as controls for HRP reporter assays.

辣根过氧化物酶（HRP）的发现是非常有利的，因为它独特的化学性质可以应用于诊断工具，包括在各种实验系统中检测氧化应激标志物，如过氧化氢（H2O2）。在这里，我们做出了令人惊讶和令人信服的发现，黄酮类化合物，根皮素和槲皮素，通常在文献中被描述为有效的抗氧化剂，强烈抑制HRP的活性。利用紫外红外（AUR）分析，我们发现低至50 μM的根皮素可以消除分离的肝脏线粒体氧化丙酮酸和苹果酸产生H2O2的检测。根皮素也使肝脏线粒体氧化琥珀酸盐或二氢乙酸盐产生的H2O2检测无效。此外，100 μM下的根皮素完全消除了AUR对H2O2的直接检测，猝灭了纯化黄嘌呤氧化酶（XO）活性的检测，但对二氯二氢荧光素（H2-DCFDA）或二氢乙胺（DHE）的荧光检测不产生干扰。剂量反应试验显示，槲皮素是一个更有效的抑制剂HRP时，与根皮素。事实上，槲皮素在nM范围内的AUR检测中消除了间苯二酚荧光，而根皮素在检测体外h2o2或分离的肝线粒体或培养的Huh-7肝癌和Mia-PaCa2细胞形成的h2o2时没有作用。总的来说，我们的研究结果表明，根皮素和槲皮素以及潜在的其他多酚类物质可能会干扰酶依赖性测定，这对设计黄酮类化合物抗氧化潜力的实验具有重要意义。我们的结果还表明，根皮素和槲皮素可以作为HRP报告检测的对照。

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引用次数: 0

Antibiotic-induced mitochondrial dysfunction: Exploring tissue-specific effects on HEI-OC1 cells and peripheral blood mononuclear cells 抗生素诱导的线粒体功能障碍：探索对HEI-OC1细胞和外周血单核细胞的组织特异性影响。

IF 2.8 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-06-11 DOI: 10.1016/j.bbagen.2025.130832

Tianshi Liu , Imen Chamkha , Eskil Elmér , Fredrik Sjövall , Johannes K. Ehinger

Antibiotics are crucial in treating infectious diseases, particularly in intensive care unit patients, but they can lead to side effects such as ototoxicity. A mechanism for this is antibiotics targeting mitochondrial components in eucaryotic cells, due to their resemblance of those in bacteria. Here we investigate how five classes of antibiotics (carbapenems, fluoroquinolones, aminoglycosides, glycopeptides, and oxazolidinones) affect mitochondrial respiratory function, ATP levels, mitochondrial membrane potential and levels of reactive oxygen species in an inner-ear derived epithelial cell line (HEI-OC1) and human primary blood cells (PBMCs) at clinically relevant concentrations.

Mitochondrial respiration in intact HEI-OC1 cells was suppressed in response to the majority of the tested antibiotics. This effect was lost when the HEI-OC1 cells were permeabilized and substrate supply controlled. Further in these cells, ROS levels were increased and ATP levels reduced. In contrast, no measure of mitochondrial function of PBMCs was affected by any antibiotics at the same concentration. We show that HEI-OC1 cells are sensitive to a broad range of antibiotics, and that the mechanism of toxicity to mitochondrial respiration is upstream of the mitochondrial respiratory chain, with downstream effects on mitochondrial respiration, ATP levels and ROS levels.

抗生素对治疗传染病至关重要，特别是对重症监护病房的病人，但它们可能导致耳毒性等副作用。其中一种机制是抗生素靶向真核细胞中的线粒体成分，因为它们与细菌中的线粒体成分相似。在这里，我们研究了五类抗生素——碳绿霉烯类、氟喹诺酮类、氨基糖苷类、糖肽类和恶唑烷酮类——在临床相关浓度下如何影响内耳上皮细胞系（HEI-OC1）和人原代血细胞（PBMCs）的线粒体呼吸功能、ATP生成、线粒体膜电位和活性氧的产生。完整的HEI-OC1细胞中的线粒体呼吸作用受到大多数抗生素的抑制。当HEI-OC1细胞渗透并控制底物供应时，这种效应就消失了。在这些细胞中，ROS生成增加，ATP水平降低。相比之下，相同浓度的抗生素对PBMCs的线粒体功能没有影响。我们发现HEI-OC1细胞对多种抗生素敏感，对线粒体呼吸的毒性机制是在线粒体呼吸链的上游，对线粒体呼吸、ATP水平和ROS水平有下游影响。

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引用次数: 0

Baicalein benefits amyotrophic lateral sclerosis via reduction of Intraneuronal misfolded protein 黄芩素通过减少神经元内错误折叠蛋白对肌萎缩性侧索硬化症有益

IF 2.8 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-06-06 DOI: 10.1016/j.bbagen.2025.130831

Chen-Hung Ting , Shao-Ting Tai , Hsiang-Yu Chang , Po-Ya Huang , Lo-Fan Cheng , Hsing-Jung Lai , Yih-Chih Kuo , Chia-Hsin Kao , I-Fan Wang , Li-Kai Tsai

Amyotrophic lateral sclerosis (ALS) is a rapidly progressive neurodegenerative disease characterized by muscle weakness and atrophy, with limited treatment options. The accumulation of misfolded proteins, such as misfolded superoxide dismutase 1 (mSOD1), contributes significantly to neuronal degeneration in ALS. Therapies targeting misfolded proteins represent a promising strategy. Baicalein, a flavonoid compound with neuroprotective properties, has shown efficacy in clearing misfolded proteins and improving behaviors in rodent models of Alzheimer's and Parkinson's diseases. However, its effects in ALS remain largely unexplored. This study demonstrated that baicalein treatment reduced total and misfolded SOD1 protein levels in both soluble and insoluble fractions of a motor neuron cell line overexpressing mutant SOD1. Baicalein also reduced intracellular SOD1 aggregates in cultured motor neurons transfected with SOD1/G93A, preserving neurite length. In an ALS mouse model expressing the SOD1/G93A transgene, baicalein treatment decreased mSOD1 aggregation, increased spinal motor neuron density, and reduced neuromuscular junction denervation. Furthermore, baicalein partially improved motor behaviors, as assessed by the rotarod test. These findings highlight baicalein's potential as a therapeutic agent for ALS, targeting intraneuronal misfolded proteins to ameliorate pathological changes and preserve motor function.

肌萎缩性侧索硬化症（ALS）是一种以肌肉无力和萎缩为特征的快速进展性神经退行性疾病，治疗选择有限。错误折叠蛋白的积累，如错误折叠的超氧化物歧化酶1 (mSOD1)，对ALS的神经元变性有重要作用。靶向错误折叠蛋白的治疗是一种很有前途的策略。黄芩素是一种具有神经保护作用的类黄酮化合物，在阿尔茨海默病和帕金森病的啮齿动物模型中显示出清除错误折叠蛋白质和改善行为的功效。然而，它对ALS的影响在很大程度上仍未被探索。该研究表明，黄芩素处理降低了过表达SOD1突变体的运动神经元细胞系的可溶性和不溶性部分的总SOD1蛋白和错误折叠的SOD1蛋白水平。黄芩苷还能降低转染SOD1/G93A的运动神经元细胞内SOD1聚集，保持神经突长度。在表达SOD1/G93A转基因的ALS小鼠模型中，黄芩苷处理降低了mSOD1聚集，增加了脊髓运动神经元密度，减少了神经肌肉接点的去神经控制。此外，黄芩素部分改善运动行为，通过旋转测试评估。这些发现突出了黄芩素作为ALS治疗药物的潜力，它可以靶向神经元内错误折叠蛋白来改善病理改变并保持运动功能。

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引用次数: 0

Tryptophan enforced alg-iron oxide nanoconjugates: A potential evalution for synergistic cancer therapy 色氨酸强化海藻氧化铁纳米缀合物：对协同癌症治疗的潜在评价

IF 2.8 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-06-04 DOI: 10.1016/j.bbagen.2025.130829

Amy Sarah Benjamin , Sunita Nayak

The contribution of magnetic nanoparticles towards the diagnosis and therapy of cancer has seen an upward graph for the past decade owing to their excellent superparamagnetic properties and contrast imaging. The surface functionalization of these iron oxide nanoparticles plays a pivotal role in the toxicity, circulation, and agglomeration parameters of clinical translation. Natural-source-based sodium alginate is a very profound biomaterial used in all areas of tissue engineering. This paper aims to study the properties of alginate-coated iron oxide nanoparticles conjugated to the aromatic amino acid tryptophan for cancer targeting. The investigation involves fabrication of the conjugated nano-system followed by analysis of the physico-chemical properties using XRD and FT-IR, evaluation of its magnetic property using VSM which shows a good superparamagnetic behaviour, along with an excellent thermal stability as shown by TGA analysis. The hyperthermia activity of these particles shows a very good specific absorption rate followed by the antioxidant property of the nano-conjugate, which shows potential scavenging activity. The biocompatibility of these nanoparticles was studied on NIH-3T3 cell lines, which showed no toxic effects, thus making the nano-conjugate an efficient cancer-targeting and therapeutic agent for future cancer nanomedicine.

磁性纳米颗粒对癌症诊断和治疗的贡献在过去十年中呈上升趋势，因为它们具有优异的超顺磁性和对比成像。这些氧化铁纳米颗粒的表面功能化在临床翻译的毒性、循环和团聚参数中起着关键作用。天然海藻酸钠是一种用途广泛的生物材料，广泛应用于组织工程的各个领域。本文旨在研究海藻酸盐包被的芳香氨基酸色氨酸共轭氧化铁纳米粒子的抗癌特性。研究包括共轭纳米体系的制备，随后使用XRD和FT-IR分析其物理化学性质，使用VSM评估其磁性能，显示出良好的超顺磁性行为，以及TGA分析显示的优异的热稳定性。这些颗粒的热疗活性显示出非常好的比吸收率，其次是纳米缀合物的抗氧化性能，显示出潜在的清除活性。在NIH-3T3细胞株上进行了生物相容性研究，结果表明纳米偶联物无毒性作用，是未来纳米肿瘤药物的有效靶向和治疗药物。

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引用次数: 0

Ferroptosis induction, androgen biosynthesis disruption and prostate cancer suppression by androgen and vitamin D combination 雄激素和维生素D联合诱导铁下垂、雄激素生物合成中断和前列腺癌抑制。

IF 2.8 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-05-28 DOI: 10.1016/j.bbagen.2025.130828

Bobae Park, Subash Khadka, Jungmi Ahn, Jacqueline Vaquiz, Okunola Igbekoyi, Chung S. Song, Bandana Chatterjee

Prostate cancer regression by hormonal vitamin D₃(calcitriol) is clinically untenable despite its well-documented anticancer effects in experimental models, since supraphysiologic dosing of calcitriol required in clinical settings causes hypercalcemia and other toxicities. We show that subnanomolar/low-nanomolar calcitriol, while non-inhibitory on its own, inhibited CRPC cells upon co-treatment with androgen (5α-DHT) at a physiologic level, evident from G1/S cell cycle arrest, induction of p21/Cip1, blockade of clonal cell growth, retardation of proliferation, migration and invasion of cells in vitro, and xenograft growth suppression in vivo. AKR1D1, an androgen-inactivating 5β-reductase, was upregulated >30-fold upon calcitriol/DHT co-treatment, while each hormone individually did not induce AKR1D1. In contrast, HSD3β1, a key enzyme for androgen biosynthesis, was upregulated >12-fold by calcitriol, which was blocked by DHT co-treatment. Elevated AKR1D1 and reduced HSD3β1 would lower intracellular androgens and contribute to CRPC repression. Autophagy mediated ferroptosis (ferritinophagy) paralleled CRPC growth inhibition by the hormone combination –indicated by LC3B induction, ferritin reduction, and upregulation of mRNAs encoding NCOA4, ALOX-5 and PTGS2 which are core participants in ferroptosis. The combination treatment caused lipid peroxidation, evident from the fluorescence shift of C11-BODIPY(581/591)-labeled cells from red to green, and from elevated malondialdehyde. GPX4 - an antioxidant peroxidase and ferroptosis regulator – was downregulated. Bipolar androgen therapy employing supraphysiologic testosterone is under clinical evaluation for efficacy against treatment-resistant metastatic CRPC, although the high androgen dose raises safety concerns. Vitamin D and androgen administered concurrently at physiologic levels may offer a superior alternative for arresting CRPC.

激素维生素D3（骨化三醇）的前列腺癌消退在临床上是站不起来的，尽管它在实验模型中具有良好的抗癌作用，因为临床需要的骨化三醇的高生理剂量会导致高钙血症和其他毒性。我们发现，亚纳摩尔/低纳摩尔骨化三醇虽然本身没有抑制作用，但在与雄激素（5α-DHT）共同作用时，在生理水平上抑制CRPC细胞，表现为G1/S细胞周期阻滞、诱导p21/Cip1、阻断克隆细胞生长、体外细胞增殖迟缓、迁移和侵袭，以及体内异种移植物生长抑制。AKR1D1是一种雄激素失活的5β-还原酶，骨化三醇/二氢睾酮共处理后，AKR1D1的表达上调了30倍，而每种激素都没有单独诱导AKR1D1。相比之下，雄激素生物合成的关键酶HSD3β1在骨化三醇的作用下上调了10倍，而DHT共处理可阻断HSD3β1的表达。升高的AKR1D1和降低的hsd3 β1会降低细胞内雄激素，并有助于抑制CRPC。自噬介导的铁细胞凋亡（铁蛋白噬）与CRPC生长抑制平行，通过激素组合-通过LC3B诱导，铁蛋白还原和编码ncoa4， ALOX-5和ptgs2的mrna上调，这些mrna是铁细胞凋亡的主要参与者。联合处理引起脂质过氧化，从C11-BODIPY（581/591）标记细胞的荧光从红色变为绿色以及丙二醛升高可以看出。GPX4 -一种抗氧化过氧化物酶和铁下垂调节因子-被下调。尽管高雄激素剂量引起了安全性问题，但采用生理上睾酮的双极雄激素治疗对治疗抵抗性转移性CRPC的疗效仍未得到临床评估。在生理水平上同时给予维生素D和雄激素可能为阻止CRPC提供更好的选择。

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引用次数: 0

Plasma-activated medium disrupts intercellular barrier function in HaCaT cells by suppressing claudin-1 expression via clathrin-dependent endocytosis 血浆活化培养基通过网格蛋白依赖内吞作用抑制claudin-1的表达，破坏HaCaT细胞间屏障功能

IF 2.8 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-05-27 DOI: 10.1016/j.bbagen.2025.130826

Chika Miyamoto , Yuta Yoshino , Hiromasa Tanaka , Hirokazu Hara , Satoshi Endo , Akira Ikari

The skin plays a critical role in protecting against water loss from the inside and pathogen invasion from the outside. The expression levels and localization of claudin-1 (CLDN1) are responsible for the tight junction (TJ) barrier function in the epidermis. Nonthermal atmospheric pressure plasma (NTAPP) has recently received attention as a novel tool in life sciences, including dermatology. NTAPP application showed useful effects on the skin, including antimicrobial activity, wound healing promotion, and anticancer activity for melanoma. However, it remains unknown how NTAPP indirect irradiation affects skin cells. In this study, we used the human epidermal keratinocyte HaCaT cells to clarify the effect of NTAPP-irradiated medium (PAM) on the epidermal TJ barrier function. Treatment with 30 % of the medium irradiated no distance from NTAPP (PAM0) significantly decreased the expression levels of CLDN1 protein. PAM0 significantly decreased the localization of CLDN1 in the cell-cell contact area. After PAM0 treatment, further culture without PAM0 significantly restored the expression and localization of CLDN1 to the same level as in the control cells. The PAM0-induced changes in protein expression and localization of CLDN1 involve lysosome degradation via a clathrin-dependent endocytosis. Treatment with PAM0 decreases transepithelial electrical resistance and increases the intercellular permeability of low-molecular-weight compounds but not high-molecular-weight compounds. The present study shows that treatment with PAM0 weakens intercellular permeability by decreasing the TJ localization of CLDN1 protein in human epidermal keratinocytes. The technology using NTAPP may be useful to promote transdermal absorption of drugs that are difficult to permeate into the body.

皮肤在防止内部水分流失和外部病原体入侵方面起着关键作用。CLDN1的表达水平和定位与表皮的紧密连接屏障功能有关。近年来，非热大气压等离子体（NTAPP）作为一种新的生命科学工具受到了包括皮肤科学在内的广泛关注。NTAPP的应用显示出对皮肤的有益作用，包括抗菌活性、促进伤口愈合和黑色素瘤的抗癌活性。然而，NTAPP间接照射对皮肤细胞的影响尚不清楚。本研究以人表皮角质形成细胞HaCaT细胞为实验材料，研究了ntapp辐照介质（PAM）对表皮TJ屏障功能的影响。与NTAPP （PAM0）无距离辐照的30%培养基显著降低了CLDN1蛋白的表达水平。PAM0显著降低了CLDN1在细胞-细胞接触区域的定位。经PAM0处理后，不加PAM0的进一步培养，CLDN1的表达和定位明显恢复到与对照细胞相同的水平。pam0诱导的CLDN1蛋白表达和定位的变化涉及溶酶体通过网格蛋白依赖的内吞作用降解。PAM0降低了经上皮电阻，增加了低分子量化合物的细胞间渗透性，但对高分子量化合物没有作用。本研究表明，PAM0通过降低人表皮角质形成细胞中CLDN1蛋白的TJ定位而减弱细胞间渗透性。使用NTAPP的技术可能有助于促进难以渗透到体内的药物的透皮吸收。

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引用次数: 0

Computational modelling of olfactory receptors 嗅觉受体的计算模型

IF 2.8 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-05-27 DOI: 10.1016/j.bbagen.2025.130825

Chiemela S. Odoemelam, Volker Steuber, Michael Schmuker

Olfactory receptors (ORs), the largest subfamily of G protein-coupled receptors, are essential for detecting and interpreting environmental odorants in animals. Understanding their function is crucial for deciphering olfactory perception and exploring emerging roles in non-olfactory systems. With the recent surge in available sequence data and AI-based structural predictions, computational modelling has become indispensable for investigating OR structure, ligand binding, and activation mechanisms. This review provides a comprehensive overview of computational approaches used in OR research, including homology modelling, molecular docking, molecular dynamics simulations, free energy calculations, pharmacophore modelling, virtual screening, and machine learning-based predictions. Both ligand-based and structure-based pharmacophore modelling are discussed in detail, highlighting their respective applications, strengths, and limitations. While structure-based approaches have gained prominence due to advances in receptor structure prediction tools like AlphaFold, ligand-based pharmacophore modelling remains valuable in scenarios where structural data are limited or uncertain. Case studies illustrate how these techniques have been applied to identify novel OR–ligand interactions, explore receptor dynamics, and support drug discovery. Collectively, these computational strategies offer powerful tools for decoding OR function, guiding experimental validation, and expanding our understanding of olfactory signalling in health and disease.

嗅觉受体（ORs）是最大的G蛋白偶联受体亚家族，对动物检测和解释环境气味至关重要。了解它们的功能对于破译嗅觉感知和探索非嗅觉系统中的新角色至关重要。随着最近可用的序列数据和基于人工智能的结构预测的激增，计算建模已经成为研究OR结构、配体结合和激活机制不可或缺的工具。本文综述了用于OR研究的计算方法，包括同源建模、分子对接、分子动力学模拟、自由能计算、药效团建模、虚拟筛选和基于机器学习的预测。本文详细讨论了基于配体和基于结构的药效团建模，重点介绍了它们各自的应用、优势和局限性。虽然基于结构的方法由于AlphaFold等受体结构预测工具的进步而获得突出地位，但基于配体的药效团建模在结构数据有限或不确定的情况下仍然有价值。案例研究说明了这些技术如何应用于识别新的or配体相互作用、探索受体动力学和支持药物发现。总的来说，这些计算策略为解码OR功能提供了强大的工具，指导实验验证，并扩展了我们对健康和疾病嗅觉信号的理解。

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引用次数: 0