Biochimica et biophysica acta. General subjects最新文献_第4页

Hepatic response to ethanol feeding in a hepatocyte-specific fatty acid binding protein-4 knock out mouse model 肝细胞特异性脂肪酸结合蛋白-4敲除小鼠模型对乙醇喂养的肝脏反应。

IF 2.2 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-11-11 DOI: 10.1016/j.bbagen.2025.130885

Neha Attal , Trenton A. Pritt , Melissa Stair , Tony E. Reeves , Iain H. McKillop

Background

Early alcohol-dependent liver disease (ALD) is characterized by increased hepatic fat storage (hepatosteatosis). Fatty acid binding protein 4 (FABP4), a protein not normally expressed in liver, becomes highly expressed in ALD. This study developed a hepatocyte-specific FABP4 mouse knockout (HS-Fabp4^−/−) to study liver responses to alcohol.

Methods

An HS-Fabp4^−/− mouse was created using a Cre/loxP embryonic stem cell approach. Male and female HS-Fabp4^−/− and wildtype (WT; C57Bl/6) mice were maintained on ethanol-drinking water (EtOH-DW) for 4-weeks. Liver damage, triglyceride content and pathology were assessed. Hepatic FABP1–9 mRNA and FABP4 and FABP5 protein were measured. Human hepatoma cell proliferation in response to exogenous FABP4 or FABP5 was analyzed.

Results

Hepatocyte-specific FABP4 deletion was confirmed in HS-Fabp4^−/− mice. No gross phenotypic differences were observed between HS-Fabp4^−/− and WT. Maintenance on EtOH-DW resulted in microsteatosis, increased hepatic triglycerides, and elevated aspartate and alanine transaminases, with no differences detected between pair-matched HS-Fabp4^−/− and WT mice. Hepatic FABP1–9 mRNA analysis revealed increased FABP4 and FABP5 mRNA expression in WT mice, and elevated FABP5 mRNA in HS-Fabp4^−/− mice in response to EtOH-DW, effects that were mirrored in serum FABP4/5 protein. Exposure of hepatoma cells to FABP4 or FABP5 revealed FABP4, but not FABP5, stimulated cell proliferation.

Conclusions

Hepatocyte-specific FABP4 deletion does not alter hepatic fat accumulation in response to EtOH feeding. Hepatic FABP4 protein produced in response to EtOH is released from hepatocytes and exogenous FABP4 promotes hepatoma cell proliferation in vitro, an effect not observed for FABP5.

背景：早期酒精依赖性肝病（ALD）的特征是肝脏脂肪储存增加（肝骨赘病）。脂肪酸结合蛋白4 (FABP4)，一种在肝脏中不正常表达的蛋白，在ALD中变得高表达。本研究开发了肝细胞特异性FABP4小鼠基因敲除（HS-Fabp4-/-）来研究肝脏对酒精的反应。方法：采用Cre/loxP胚胎干细胞法建立HS-Fabp4-/-小鼠。雄性和雌性HS-Fabp4-/-和野生型（WT; C57Bl/6）小鼠在乙醇饮用水（EtOH-DW）中维持4周。评估肝损害、甘油三酯含量及病理。测定肝脏FABP1-9 mRNA和FABP4、FABP5蛋白表达。分析外源性FABP4或FABP5对人肝癌细胞增殖的影响。结果：在HS-Fabp4-/-小鼠中证实肝细胞特异性FABP4缺失。HS-Fabp4-/-和WT之间没有明显的表型差异。维持EtOH-DW导致微脂肪变性、肝脏甘油三酯升高、天冬氨酸和丙氨酸转氨酶升高，配对的HS-Fabp4-/-和WT小鼠之间没有差异。肝脏FABP1-9 mRNA分析显示，在WT小鼠中FABP4和FABP5 mRNA表达升高，在HS-Fabp4-/-小鼠中FABP5 mRNA表达升高，这反映在血清FABP4/5蛋白中。肝癌细胞暴露于FABP4或FABP5后，发现FABP4刺激细胞增殖，而FABP5没有。结论：肝细胞特异性FABP4缺失不会改变EtOH喂养后肝脏脂肪的积累。在体外实验中，外源性FABP4可促进肝癌细胞增殖，而FABP5则未观察到这种作用。

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引用次数: 0

Pharmacological inhibition of Trypanosoma cruzi aldo-keto reductase (TcAKR) and its effect on benznidazole resistance 克氏锥虫醛酮还原酶的药理抑制及其对苯并硝唑耐药性的影响

IF 2.2 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-11-08 DOI: 10.1016/j.bbagen.2025.130880

Patricia Andrea Garavaglia , Sebastián Aduviri , Pablo Trujillo , Laura Mónica Tasso , Joaquín Juan Bautista Cannata , Monica Pickholz , Gabriela Andrea García

Chagas disease, caused by the protozoan Trypanosoma cruzi, has become a global health concern due to increased globalization. Several studies suggest that the aldo-keto reductase from T. cruzi (TcAKR) is involved in resistance to benznidazole, the drug commonly used to treat this infection.

To further support the role of TcAKR in drug resistance, we evaluated its interaction with four compounds —quercetin, flufenamic acid, phenolphthalein, and menadione—previously reported as inhibitors of other AKRs. Molecular docking was performed to assess affinity and molecular specific interactions, and the inhibitory effects of these compounds on both TcAKR activities —aldo-keto reductase and quinone-oxidoreductase— were experimentally determined.

Binding affinities, in decreasing order, were: quercetin > flufenamic acid > phenolphthalein > menadione. Both quercetin and flufenamic acid interacted with amino acid residues located outside the enzyme's active site. Quercetin completely inhibited both TcAKR activities, while flufenamic acid inhibited approximately 50 %. Phenolphthalein and menadione showed low levels of inhibition. The inhibition profiles of quercetin and flufenamic acid were consistent with a noncompetitive mechanism.

The effect of quercetin on benznidazole resistance was evaluated in transfected parasites overexpressing TcAKR, which are 1.8-fold more resistant to this drug. Quercetin treatment restored benznidazole sensitivity in these parasites, reducing the IC₅₀ to levels comparable to those of control parasites. These results provide further evidence of TcAKR's involvement in benznidazole resistance and suggest that its inhibition can enhance treatment efficacy.

恰加斯病是由原生动物克氏锥虫引起的，由于全球化的加剧，该病已成为一个全球性的健康问题。几项研究表明，克氏锥虫的醛酮还原酶（TcAKR）参与了对苯并硝唑的耐药性，苯并硝唑是通常用于治疗这种感染的药物。为了进一步支持TcAKR在耐药中的作用，我们评估了它与四种化合物的相互作用——槲皮素、氟芬那酸、酚酞和美萘醌——之前被报道为其他akr的抑制剂。通过分子对接来评估亲和力和分子特异性相互作用，并通过实验确定了这些化合物对TcAKR活性-醛酮还原酶和醌氧化还原酶的抑制作用。结合亲和度由高到低依次为：槲皮素；氟胺酸；酚酞；美萘酮。槲皮素和氟芬那酸都与酶活性位点外的氨基酸残基相互作用。槲皮素完全抑制两种taccr活性，而氟芬那酸抑制约50%。酚酞和美萘醌的抑制作用较低。槲皮素和氟芬那酸的抑制作用与非竞争性机制一致。研究了槲皮素对过表达TcAKR的疟原虫对苯并硝唑耐药性的影响，结果表明槲皮素对该药物的耐药性提高了1.8倍。槲皮素处理恢复了这些寄生虫对苯并硝唑的敏感性，将IC₅0降低到与对照寄生虫相当的水平。这些结果进一步证明了TcAKR参与苯并硝唑耐药，并提示其抑制可以提高治疗效果。

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引用次数: 0

Nanobody-based imaging: Advancing precision in molecular diagnostics 基于纳米体的成像：提高分子诊断的精度。

IF 2.2 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-11-08 DOI: 10.1016/j.bbagen.2025.130884

Zohre Eftekhari, Fatemeh Kazemi-Lomedasht

Molecular imaging is a cornerstone of modern medicine, enabling non-invasive visualization of biological processes at the molecular level. The emergence of nanobodies (Nbs), small single-domain antibody fragments derived from camelids, has transformed this field due to their superior tissue penetration, rapid clearance, and high target specificity compared to conventional antibodies. This review focuses on the integration of Nbs with positron emission tomography (PET) and single-photon emission computed tomography (SPECT) two complementary molecular imaging modalities known for their high sensitivity, quantitative potential, and clinical relevance. Nb-based PET and SPECT imaging probes are emerging as powerful tools for detecting disease-associated molecular targets with exceptional precision. Their unique properties, including high affinity, specificity, and stability, make them ideal candidates for developing advanced radiotracers that enable early disease detection, monitoring of therapeutic responses, and evaluation of novel treatment strategies. Despite these advantages, several challenges remain, such as scalable Nb production, reduction of immunogenicity in clinical applications, and optimization of radiolabeling methods that preserve Nb integrity and function. This review highlights recent advances in Nb engineering, radiolabeling strategies, and preclinical and clinical applications of Nb-based PET and SPECT imaging, while outlining critical directions for future research. By addressing current limitations, Nb-based molecular imaging holds great promise for improving diagnostic accuracy, advancing personalized medicine, and expanding its clinical impact across diverse disease areas.

分子成像是现代医学的基石，能够在分子水平上对生物过程进行无创可视化。纳米体（Nbs）的出现，源自骆驼的小单域抗体片段，由于其与传统抗体相比具有优越的组织穿透性，快速清除和高靶向特异性，已经改变了这一领域。这篇综述的重点是Nbs与正电子发射断层扫描（PET）和单光子发射计算机断层扫描（SPECT）的整合，这两种互补的分子成像方式以其高灵敏度、定量潜力和临床相关性而闻名。基于nb的PET和SPECT成像探针正在成为检测疾病相关分子靶标的强大工具，具有极高的精度。它们独特的特性，包括高亲和力、特异性和稳定性，使它们成为开发先进放射性示踪剂的理想候选者，可以用于早期疾病检测、监测治疗反应和评估新的治疗策略。尽管有这些优势，但仍存在一些挑战，例如可扩展的铌生产，临床应用中免疫原性的降低，以及保持铌完整性和功能的放射性标记方法的优化。本文综述了铌工程、放射性标记策略以及基于铌的PET和SPECT成像的临床前和临床应用方面的最新进展，同时概述了未来研究的关键方向。通过解决当前的局限性，基于nb的分子成像在提高诊断准确性、推进个性化医疗和扩大其在不同疾病领域的临床影响方面具有很大的前景。

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引用次数: 0

Structural characterization of a L-dehydroascorbic acid–L-homocysteine thiolactone reaction product: Intracellular formation in neuronal cells l -脱氢抗坏血酸- l -同型半胱氨酸硫内酯反应产物的结构表征：神经元细胞内形成

IF 2.2 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-11-05 DOI: 10.1016/j.bbagen.2025.130882

Ghizlane Loubane , Gabriel Robert , Syed Benazir Firdaus , Raphaël Robidas , Christian Comeau , Pierre-Luc Boudreault , Jeampy E. Komba , Hugo Gagnon , J Richard Wagner , Stephen Naylor , Klaus Klarskov

Homocysteine thiolactone (HTL) has been implicated in cardiovascular and neurological pathologies. While homocysteine can S-homocysteinylate thiol groups in proteins, the chemical properties of HTL facilitates covalent binding to protein ε-amino groups on lysine residues, which can initiate protein aggregation. Ascorbate plays an important role in the prevention of oxidative stress. Ascorbate is easily oxidized by the loss of two electrons to dehydroascorbate (DHA), which can be reduced back to ascorbate by thiol-containing smaller and larger molecules. In the present study, reaction products between two aldehydes, formaldehyde and propionaldehyde as well as DHA with the physiological relevant isomer of homocysteine thiolactone i.e. L-HTL were purified, and their structure was determined by 1D and 2D-nuclear magnetic resonance. In all three cases the reaction products are likely formed by initial imine condensation, subsequent formation of a hemiaminal product followed by HTL ring opening and intramolecular nucleophilic attack of the resulting thiol anion to form a six-member thiazinane ring with a carboxylic acid group. The structure of the DHA, L-HTL reaction product was confirmed by high resolution accurate ESIMS/MS in negative mode. Formation of the reaction product between DHA and HTL prevented N-homocysteinylation of cytochrome c by HTL, confirming earlier observations. The reaction product is formed in human neuroblastoma cells (SH-SY5Y) when exposed to HTL and DHA or ascorbate, potentially preventing protein aggregation. The consequences associated with formation of a reaction product between DHA and HTL suggest that DHA could protect against protein N-homocysteinylation.

同型半胱氨酸硫内酯（HTL）与心血管和神经系统疾病有关。同型半胱氨酸可以使蛋白质中的巯基s -同型半胱氨酸化，而HTL的化学性质有助于与赖氨酸残基上的蛋白质ε-氨基共价结合，从而引发蛋白质聚集。抗坏血酸在预防氧化应激中起着重要作用。抗坏血酸很容易因失去两个电子而氧化为脱氢抗坏血酸（DHA），后者可以通过含有较小和较大硫醇的分子还原为抗坏血酸。本研究纯化了甲醛与丙醛两种醛以及DHA与同型半胱氨酸硫内酯生理相关异构体L-HTL的反应产物，并通过1D和2d核磁共振测定了它们的结构。在所有三种情况下，反应产物可能是由最初的亚胺缩合形成，随后形成半胺产物，然后是HTL环打开和产生的硫醇阴离子的分子内亲核攻击，形成具有羧基的六元噻嗪环。采用高精度ESIMS/MS负模式对反应产物DHA、L-HTL的结构进行了确证。DHA和HTL之间的反应产物的形成阻止了HTL对细胞色素c的n -同型半胱氨酸化，证实了早期的观察。当暴露于HTL和DHA或抗坏血酸时，反应产物在人神经母细胞瘤细胞（SH-SY5Y）中形成，可能阻止蛋白质聚集。DHA和HTL之间形成反应产物的结果表明，DHA可以防止蛋白质n -同型半胱氨酸化。

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引用次数: 0

Screening of biomarkers in autoimmune thyroid diseases (AITDs): preliminary study based on Raman spectroscopy and MALDI-ToF mass spectrometry 自身免疫性甲状腺疾病（AITDs）生物标志物的筛选：基于拉曼光谱和MALDI-ToF质谱的初步研究

IF 2.2 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-11-05 DOI: 10.1016/j.bbagen.2025.130883

Sara Trzos , Sylwia Orzechowska , Paweł Link-Lenczowski , Grzegorz Sokołowski , Malgorzata Baranska , Ewa Pocheć

Hashimoto's thyroiditis (HT) and Graves' disease (GD), autoimmune thyroid diseases (AITDs), are among the most commonly reported autoimmune disorders. Early and unambiguous diagnosis and monitoring of the disease development are crucial to obtain the most effective treatment results. Matrix-assisted laser desorption/ionization with a time-of-flight analyzer mass spectrometry (MALDI-ToF MS) and Raman spectroscopy are promising analytical methods in this regard. These non-invasive and sensitive techniques provide information on specific features of proteins, including their post-translational modifications and the level of a wide range of biomolecules in the bloodstream, which are useful for assessing the health status of patients. This study screened for potential biomarkers in AITDs using both methods. Sera from HT patients at two stages of disease progression and GD patients before and after normalization of thyrotropic hormone following immunosuppressive treatment were used in the study. Serum biomolecule changes were analyzed using Raman spectroscopy. N-glycans released from serum glycoproteins were detected by MALDI-ToF mass spectrometry. We observed that the levels of phenylalanine, carotenoids, and phospholipids in HT sera correlate with increased inflammation accompanying this disease. The GD group showed a lower amount of serum collagen compared to the HT patients. Quantitative comparison of N-glycans revealed several differences between the study groups and healthy donors. The higher galactosylation and α2,6-sialylation of serum proteins in HT2 relative to GD patients are the main differences in N-glycan profiles of AITDs. The obtained preliminary results demonstrate that these analytical techniques have potential in diagnosing and monitoring AITDs, and can be a good alternative to the currently used methods. Analysis of a larger number of samples, as well as a more detailed MS methodology for precise decoding of glycan structures, is necessary for further research.

桥本甲状腺炎（HT）和格雷夫斯病（GD），自身免疫性甲状腺疾病（AITDs），是最常报道的自身免疫性疾病。早期和明确的诊断和监测疾病发展是至关重要的，以获得最有效的治疗结果。基质辅助激光解吸/电离与飞行时间分析仪质谱（MALDI-ToF MS）和拉曼光谱是这方面有前途的分析方法。这些非侵入性和敏感的技术提供了蛋白质的特定特征信息，包括它们的翻译后修饰和血液中各种生物分子的水平，这对评估患者的健康状况很有用。本研究使用这两种方法筛选AITDs的潜在生物标志物。本研究采用HT患者两期病程和GD患者经免疫抑制治疗后促甲状腺激素恢复正常前后的血清。用拉曼光谱分析血清生物分子变化。采用MALDI-ToF质谱法检测血清糖蛋白释放的n -聚糖。我们观察到HT血清中苯丙氨酸、类胡萝卜素和磷脂的水平与此病伴随的炎症增加相关。GD组血清胶原蛋白含量低于HT组。n -聚糖的定量比较揭示了研究组与健康供者之间的一些差异。与GD患者相比，HT2患者血清蛋白的半乳糖基化和α2,6-唾液基化水平较高是AITDs n -聚糖谱的主要差异。初步结果表明，这些分析技术在诊断和监测AITDs方面具有一定的潜力，可以作为现有方法的一个很好的替代方案。进一步的研究需要分析大量的样品，以及更详细的质谱方法来精确解码多糖结构。

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引用次数: 0

The role of ATM in sodium butyrate-mediated inhibition of macrophage polarization ATM在丁酸钠介导的巨噬细胞极化抑制中的作用。

IF 2.2 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-11-04 DOI: 10.1016/j.bbagen.2025.130881

Yan-Yan Guo, Xia-Nan Chu, Pei-Hua Wang, Ya-Qian Li, Li Xing

DNA damage response (DDR) signaling not only maintains genomic integrity but also plays a role in the activation of immune cells, including macrophages. In response to the stimuli, macrophages can be polarized into a proinflammatory phenotype, M1. In the monocytoid THP-1 cell-derived macrophage model, sodium butyrate was found to inhibit the expression of M1 biomarkers TNF-α, IL-6, IL-1β, and CXCL10, and downregulate DDR-associated proteins, including the apical Ataxia-Telangiectasia mutated protein kinase (ATM). However, siRNA-mediated knockdown of ATM did not reduce the expression of M1 biomarkers, but still downregulated the expression of DDR-associated proteins such as RAD50, p53, CHK1, NBS1, and γH2AX. Moreover, ATM knockdown modulated the expression of innate immunity regulatory genes, including sialic acid binding immunoglobulin type lectins 14 (Siglec14), Siglec15, signaling lymphocyte activation molecule family 1 (Siamf1), Siamf7, and guanylate binding protein 2 in response to the infection of Mycobacterium tuberculosis H37Ra. The results suggest that ATM may serve as a regulator to couple the DDR and innate immune response of macrophages, but barely contributes to the sodium butyrate-mediated inhibition of certain M1 biomarkers.

DNA损伤反应（DDR）信号不仅维持基因组的完整性，而且在包括巨噬细胞在内的免疫细胞的激活中发挥作用。在对刺激的反应中，巨噬细胞可以极化成促炎表型M1。在单核细胞THP-1细胞来源的巨噬细胞模型中，发现丁酸钠抑制M1生物标志物TNF-α、IL-6、IL-1β和CXCL10的表达，下调ddr相关蛋白，包括顶端失调性毛细血管扩张突变蛋白激酶（ATM）。然而，sirna介导的ATM敲低并不会降低M1生物标志物的表达，但仍会下调ddr相关蛋白如RAD50、p53、CHK1、NBS1和γH2AX的表达。此外，ATM敲低可调节天然免疫调节基因的表达，包括唾液酸结合免疫球蛋白型凝集素14 （Siglec14）、Siglec15、信号淋巴细胞激活分子家族1 （Siamf1）、Siamf7和鸟苷酸结合蛋白2，以响应结核分枝杆菌H37Ra感染。结果表明，ATM可能作为巨噬细胞DDR和先天免疫应答的调节因子，但几乎不参与丁酸钠介导的对某些M1生物标志物的抑制。

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引用次数: 0

Copper transporter 1 contributes to the progression of cholangiocarcinoma 铜转运蛋白1参与胆管癌的进展。

IF 2.2 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-11-02 DOI: 10.1016/j.bbagen.2025.130876

Yanpeng Ma , Longlong Liu , Jiayue Duan , Xiaojian Wang

Background

Cholangiocarcinoma (CCA) is characterized by poor prognosis and a lack of effective biomarkers and therapeutic targets. Although copper dysregulation has been implicated in CCA, the role of the copper transporter SLC31A1 (CTR1) in its progression remains poorly understood.

Methods

The mRNA and protein levels of SLC31A1 in tumor and adjacent non-tumor tissues was evaluated by RT-PCR and enzyme-linked immunosorbent assay, respectively. Tissue copper concentrations were determined using a colorimetric complexation assay. Correlations between SLC31A1 expression, copper accumulation, and clinicopathological parameters were evaluated using Pearson's analysis. Cell proliferation and colony formation were assessed by the cell counting kit 8 and colony formation assays, respectively. Mitochondrial function was assessed by measuring intracellular ATP levels, reactive oxygen species (ROS) production, and the glutathione redox ratio.

Results

Elevated SLC31A1 expression was significantly associated with microvascular invasion, larger tumor size (≥5 cm), and advanced AJCC stage (III–IV). Both SLC31A1 mRNA and protein levels were significantly upregulated in CCA tissues compared with adjacent non-tumor tissues and correlated with copper accumulation. In vitro, SLC31A1 was highly expressed across multiple CCA cell lines. Silencing SLC31A1 in RBE cells impaired cell proliferation and colony formation. Mechanistically, depletion of SLC31A1 increased intracellular ROS, reduced the GSH/GSSG ratio, and decreased ATP production, indicating disruption of mitochondrial function and redox homeostasis.

Conclusions

SLC31A1 promotes cholangiocarcinoma progression by maintaining copper homeostasis, mitochondrial integrity, and redox balance. These findings suggest that SLC31A1 may serve as a potential prognostic biomarker and a candidate therapeutic target.

背景：胆管癌（CCA）的特点是预后差，缺乏有效的生物标志物和治疗靶点。尽管铜的失调与CCA有关，但铜转运体SLC31A1 （CTR1）在其进展中的作用仍然知之甚少。方法：分别采用RT-PCR和酶联免疫吸附法检测肿瘤组织和癌旁非肿瘤组织中SLC31A1 mRNA和蛋白水平。用比色络合法测定组织铜浓度。使用Pearson分析评估SLC31A1表达、铜积累和临床病理参数之间的相关性。分别用细胞计数试剂盒8和菌落形成试验评估细胞增殖和菌落形成。通过测量细胞内ATP水平、活性氧（ROS）产生和谷胱甘肽氧化还原比来评估线粒体功能。结果：SLC31A1表达升高与微血管侵犯、较大肿瘤大小（≥5 cm）、晚期AJCC （III-IV）相关。与邻近非肿瘤组织相比，CCA组织中SLC31A1 mRNA和蛋白水平均显著上调，并与铜积累相关。在体外，SLC31A1在多个CCA细胞系中高表达。在RBE细胞中沉默SLC31A1会损害细胞增殖和集落形成。从机制上讲，SLC31A1的缺失增加了细胞内ROS，降低了GSH/GSSG比率，减少了ATP的产生，表明线粒体功能和氧化还原稳态受到破坏。结论：SLC31A1通过维持铜稳态、线粒体完整性和氧化还原平衡来促进胆管癌的进展。这些发现表明，SLC31A1可能作为潜在的预后生物标志物和候选治疗靶点。

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引用次数: 0

Rationally designed peptide induces apoptosis and cell cycle modulation in resistant melanoma 合理设计多肽诱导耐药黑色素瘤细胞凋亡和细胞周期调节。

IF 2.2 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-11-02 DOI: 10.1016/j.bbagen.2025.130877

Layza Sá Rocha , Ana Cristina Jacobowski , Eduarda Thiburcio , Rafael Araujo Pereira , Claudiane Vilharroel Almeida , Camila De Oliveira Gutierrez , Thaís de Andrade Farias Rodrigues , Rodrigo Juliano Oliveira , Gabriel B. Taveira , Priscila Aiko Hiane , Ana Paula de Araújo Boleti , Octávio Luiz Franco , Marlon Henrique Cardoso , Maria Ligia Rodrigues Macedo

The ineffectiveness of conventional therapies against melanoma necessitates the development of novel approaches characterized by high selectivity and low systemic toxicity. Using computational optimization, KW18, a synthetic α-helical peptide, was rationally engineered. It incorporates conserved bioactive motifs from patented peptides. The Joker algorithm was used to refine the peptide. This process enhanced its positive charge, hydrophobicity, and α-helical stability. In vitro, KW18 displayed potent cytotoxicity against melanoma cells (IC₅₀ = 11.41–21.75 μM) with 84–91 % inhibition at 128 μM, minimal toxicity to FN1 fibroblasts, hemolysis below 5 %, and stability exceeding 80 %. Functional assays revealed mitochondrial damage and caspase-dependent apoptosis (86.5 % late apoptosis at 24 h). Cell cycle analysis in B16F10-Nex2 cells revealed a moderate accumulation in G0/G1 phase (62.9 %) alongside a substantial S-phase population (25.8 %), with minimal Sub-G0 (5.2 %) and reduced G2/M (6.1 %). This profile suggests a cytostatic-like effect through partial cell cycle slowdown. Structural characterization via helical wheel projections and circular dichroism confirmed its α-helical conformation, consistent with membrane-targeting activity. In vivo, Galleria mellonella larvae exposed to KW18 exhibited 100 % survival, supporting a favorable safety profile. Collectively, KW18 combines apoptosis induction with cell cycle modulation, offering a dual mechanism against melanoma while sparing healthy cells. These findings designate KW18 as a stable, selective, and safe therapeutic option for drug-resistant melanoma and a beneficial element for combined treatments.

传统治疗方法对黑色素瘤无效，需要开发具有高选择性和低全身毒性的新方法。通过计算优化，对合成α-螺旋肽KW18进行了合理的工程化设计。它结合了来自专利肽的保守的生物活性基序。使用Joker算法对肽进行细化。该工艺增强了其正电荷性、疏水性和α-螺旋稳定性。在体外，KW18对黑色素瘤细胞（IC₅₀ = 11.41-21.75 μM）显示出强大的细胞毒性，在128 μM处抑制84-91 %，对FN1成纤维细胞的毒性最小，溶血率低于5 %，稳定性超过80 %。功能分析显示线粒体损伤和caspase依赖性凋亡（24 h时86.5 %晚期凋亡）。B16F10-Nex2细胞的细胞周期分析显示，在G0/G1期（62.9 %）和大量s期群体（25.8 %）中有适度的积累，亚G0期（5.2 %）最小，G2/M降低（6.1 %）。这表明细胞静态样效应通过部分细胞周期减慢。通过螺旋轮投影和圆二色性的结构表征证实了其α-螺旋构象，与膜靶向活性一致。在体内，暴露于KW18的mellongalleria幼虫的存活率为100% %，支持良好的安全性。总的来说，KW18结合了细胞凋亡诱导和细胞周期调节，在保护健康细胞的同时提供了抗黑色素瘤的双重机制。这些发现表明KW18是一种稳定、选择性和安全的耐药黑色素瘤治疗选择，也是联合治疗的有益元素。

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引用次数: 0

Elevated cyclic hydrostatic pressure enhances the transfection activity of lipoplexes by activating clathrin-mediated endocytosis 升高的循环静水压力通过激活网格蛋白介导的内吞作用来增强脂丛的转染活性。

IF 2.2 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-11-01 DOI: 10.1016/j.bbagen.2025.130878

Weichen Zhan , Xiaowei Ding , Zhongrui Cui , Yizhuo Wu , Yiwen Gu , Hanxiao Cheng , Xinxin Ge , Yun Wang , Jiangyun Luo , Bing Xiao

Despite significant advancements in liposome-mediated transfection technology over the past decades, achieving optimal transfection efficiency with lipoplex remains challenging in certain primary cells, such as vascular smooth muscle cells, endothelial cells, and suspension cells. Here, we present an innovative approach to significantly enhance Lipofectamine-based transient transfection efficiency in hard-to-transfect cells by applying elevated cyclic hydrostatic pressure (CHP). The plasmids encoding the enhanced green fluorescent protein (EGFP) were transfected using Lipofectamine 3000 reagent, and the transfection efficiency was evaluated by Western blot or flow cytometry. Our results demonstrate that CHP (0.0083 Hz, 0–100 mmHg) significantly enhanced the transfection efficiency of lipoplex in primary human aortic smooth muscle cells (HASMCs) and other difficult-to-transfect cell types. Mechanistic studies revealed that the enhancement of liposome-mediated transfection by CHP was dependent on the activation of clathrin-dependent endocytic pathways. Importantly, this mechanical stimulation did not affect the proliferative or migratory capacities of HASMCs, despite the identification of significantly modulated proteins (5.8 % of the total proteome) by proteomic analysis. This study establishes a novel, safe strategy to enhance lipoplex-mediated nucleic acid delivery in challenging-to-transfect cell types.

尽管在过去的几十年里，脂质体介导的转染技术取得了重大进展，但在某些原代细胞中，如血管平滑肌细胞、内皮细胞和悬浮细胞，脂质体的转染效率仍然具有挑战性。在这里，我们提出了一种创新的方法，通过提高循环静水压力（CHP），显著提高脂蛋白胺在难以转染的细胞中的瞬时转染效率。用Lipofectamine 3000试剂转染编码增强绿色荧光蛋白（EGFP）的质粒，用Western blot或流式细胞术评价转染效率。我们的研究结果表明，CHP（0.0083 Hz, 0-100 mmHg）显著提高了脂质体在人主动脉平滑肌细胞（HASMCs）和其他难以转染的细胞类型中的转染效率。机制研究表明，脂质体介导的CHP转染的增强依赖于网格蛋白依赖的内吞途径的激活。重要的是，尽管通过蛋白质组学分析鉴定出显著调节的蛋白质（占总蛋白质组的5.8% %），但这种机械刺激并未影响HASMCs的增殖或迁移能力。本研究建立了一种新的、安全的策略来增强脂质体介导的核酸在挑战转染细胞类型中的传递。

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引用次数: 0

Hyperglycemia accelerated the metastasis of triple-negative breast cancer via promoting TNFα/Gli-1 axis in endothelial cells 高血糖通过促进内皮细胞中TNFα/ gli1轴的表达加速三阴性乳腺癌的转移。

IF 2.2 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. General subjects

Pub Date : 2025-11-01 DOI: 10.1016/j.bbagen.2025.130875

Xiyu Mei , Chuang Ke , Ziyun Gao , Fan Yang , Zhenlin Huang , Bin Lu , Lili Ji

Diabetes mellitus (DM) is associated with a poor prognosis of aggressive breast cancer. Vascular dysfunction is commonly found during the development of both cancer and diabetes. We previously reported that the disruption of vascular endothelial phenotype induced by tumor necrosis factor-α (TNFα) accelerated the trans-endothelial metastasis of triple-negative breast cancer (TNBC). Herein, we explored the role of vascular endothelial cells in diabetes-induced TNBC metastasis. Both type 2 DM (T2DM) and type 1 DM (T1DM) enhanced the metastasis of TNBC in vivo. T2DM increased the expression of endothelial phenotype vascular endothelial cadherin (VE-cadherin), platelet-endothelial cell adhesion molecule (PECAM-1/CD31), and mesenchymal markers including vimentin and fibroblast specific protein-1 (FSP-1/S100A4) in tumor vessels. T1DM increased the expression of vimentin and FSP-1, but suppressed the expression of VE-cadherin in tumor vessels. Hyperglycemia elevated the production of TNFα in vivo and in vitro. TNFα reduced the trans-endothelial electrical resistance (TEER) value of both human mammary microvascular endothelial cells (HMMECs) and human umbilical vein endothelial cells (HUVECs). Expressions of vimentin and α-smooth muscle actin (α-SMA) were also increased in TNFα-treated both HMMECs and HUVECs. The number of trans-endothelial migrated MDA-MB-231 cells through TNFα-treated HMMECs or HUVECs monolayer was elevated. Moreover, glioma-associated oncogene 1 (Gli-1) was remarkably accumulated in the nucleus of TNFα-stimulated HMMECs and DM-induced tumor vessels. Both Gli-1 siRNA and GANT61 (an inhibitor of Gli-1) could abrogate the increased TNBC trans-endothelial migration through TNFα-treated ECs. We demonstrated that DM might promote TNBC metastasis via activating the TNFα/Gli-1 axis initiated vascular endothelial mesenchymal-like phenotype.

糖尿病（DM）与侵袭性乳腺癌预后不良相关。血管功能障碍在癌症和糖尿病的发展过程中都很常见。我们之前报道过肿瘤坏死因子-α (tnf -α)诱导的血管内皮表型破坏加速了三阴性乳腺癌（TNBC）的跨内皮转移。在此，我们探讨了血管内皮细胞在糖尿病诱导的TNBC转移中的作用。2型糖尿病（T2DM）和1型糖尿病（T1DM）均可促进TNBC在体内的转移。T2DM增加了内皮型血管内皮钙粘蛋白（VE-cadherin）、血小板-内皮细胞粘附分子（PECAM-1/CD31）以及血管entin和成纤维细胞特异性蛋白-1 （FSP-1/S100A4）等间充质标志物在肿瘤血管中的表达。T1DM增加了vimentin和FSP-1的表达，抑制了VE-cadherin在肿瘤血管中的表达。在体内和体外，高血糖升高了TNFα的产生。TNFα降低了人乳腺微血管内皮细胞（hmmes）和人脐静脉内皮细胞（HUVECs）的跨内皮电阻（TEER）值。在tnf α处理的hmmes和huvec中，波形蛋白和α-平滑肌肌动蛋白（α-SMA）的表达也增加。通过tnf α处理的hmmec或huvec单层，跨内皮迁移的MDA-MB-231细胞数量增加。此外，胶质瘤相关癌基因1 （gli1）在tnf α刺激的hmmec和dm诱导的肿瘤血管的细胞核中显著积累。gli1 siRNA和GANT61（一种gli1抑制剂）都可以通过tnf α处理的内皮细胞消除TNBC跨内皮迁移的增加。我们证明了糖尿病可能通过激活TNFα/ gli1轴启动的血管内皮间充质样表型来促进TNBC转移。

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引用次数: 0