Biochimica et biophysica acta. Molecular cell research最新文献_第2页

Hornerin expressed on endothelial cells via interacting with thrombomodulin modulates vascular inflammation and angiogenesis 内皮细胞上表达的角蛋白通过与血栓调节蛋白相互作用调节血管炎症和血管生成。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-02-01 DOI: 10.1016/j.bbamcr.2024.119891

Takayuki Okamoto , Mai Hattori , Yukiko Katsube , Junichi Ota , Kunihiro Asanuma , Haruki Usuda , Koichiro Wada , Koji Suzuki , Tetsuro Nikai

Thrombomodulin is predominantly expressed on vascular endothelial cells and modulates endothelial cell functions by interacting with multiple ligands. The specific thrombomodulin receptor or cofactor active on the endothelial cell surface remains elusive. This study aims to identify interacting partners of thrombomodulin on endothelial cells. Here, using a liquid chromatograph-tandem mass spectrometer, hornerin was identified as a candidate protein. We then investigated hornerin protein and mRNA expression in endothelial cells. Hornerin protein was detected in the mouse endothelium of the aorta and lung. Both human- and mouse-cultured endothelial cells expressed hornerin mRNA and protein. Moreover, immunoprecipitation analysis suggested the direct protein interaction between thrombomodulin and hornerin. Lipopolysaccharides administration increased serum hornerin concentrations in mice and reduced hornerin protein levels on the surface of cultured endothelial cells as same as thrombomodulin protein. Thrombomodulin-targeting siRNA decreased not only thrombomodulin protein levels but also hornerin protein levels in cultured endothelial cells. Thrombomodulin- or hornerin-targeting siRNA impaired tube formation and leukocyte adhesion to endothelial cells. Our findings reveal that hornerin is located on vascular endothelial cells in the presence of thrombomodulin and suggest that endothelial thrombomodulin and hornerin may interact, which may play an important role in endothelial cell functions such as vascular inflammation and angiogenesis.

血栓调节蛋白主要在血管内皮细胞上表达，并通过与多种配体相互作用调节内皮细胞功能。内皮细胞表面特定的血栓调节素受体或辅助因子活性尚不明确。本研究旨在确定血栓调节素对内皮细胞的相互作用伙伴。在这里，使用液相色谱-串联质谱仪，角蛋白被确定为候选蛋白。然后我们研究了内皮细胞中角蛋白和mRNA的表达。在小鼠主动脉和肺内皮中检测到Hornerin蛋白。人和小鼠内皮细胞均表达角蛋白mRNA和蛋白。此外，免疫沉淀分析表明血栓调节蛋白和角蛋白之间存在直接的蛋白相互作用。脂多糖增加小鼠血清角蛋白浓度，降低培养内皮细胞表面角蛋白水平，与血栓调节蛋白水平相同。以血栓调节蛋白为靶点的siRNA不仅能降低培养内皮细胞的血栓调节蛋白水平，还能降低角蛋白水平。以血栓调节素或角蛋白为靶点的siRNA会破坏小管的形成和白细胞与内皮细胞的粘附。我们的研究结果表明，在血栓调节蛋白存在的情况下，角蛋白位于血管内皮细胞上，表明内皮血栓调节蛋白和角蛋白可能相互作用，在血管炎症和血管生成等内皮细胞功能中发挥重要作用。

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引用次数: 0

Enhanced radiotherapy susceptibility in NSCLC through palbociclib-mediated PP5 inhibition 通过帕博西利介导的PP5抑制增强NSCLC的放疗敏感性。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-02-01 DOI: 10.1016/j.bbamcr.2024.119884

Chao-Yuan Huang , Li-Ju Chen , Grace Chen , Cheng-Yi Wang , Shiao-Ya Hong

Radiotherapy remains a cornerstone in the treatment of non-small cell lung cancer (NSCLC), yet radioresistance often limits its efficacy. Identifying molecular targets that enhance radiosensitivity is crucial to offering both curative and palliative benefits for patients with NSCLC. Utilizing bioinformatics analysis, our study revealed significantly higher expression of PP5 in NSCLC tissues compared to normal tissues. Kaplan-Meier survival analysis also showed that high PP5 expression correlates with poorer overall survival, particularly in patients undergoing radiotherapy, suggesting a role for PP5 in radioresistance. We further demonstrated that PP5 is a critical target of palbociclib, distinct from CDK4/6, influencing radiosensitivity in NSCLC. Palbociclib enhanced radiotherapy susceptibility by inducing sustained DNA damage and AMPK activation. The subsequent cellular event is apoptosis rather than autophagy. Furthermore, the enhanced efficacy of combination therapy was counteracted by an AMPK inhibitor and PP5 activator, underscoring the importance of these pathways in mediating the response. Our findings provide compelling evidence that targeting PP5 can significantly enhance the therapeutic outcomes of radiotherapy in NSCLC. This research offers valuable insights into new combination therapy strategies, highlighting the potential of PP5 as a novel therapeutic target to overcome radioresistance.

放疗仍然是治疗非小细胞肺癌（NSCLC）的基石，但放射耐药往往限制了其疗效。确定增强放射敏感性的分子靶点对于为非小细胞肺癌患者提供治疗和姑息效益至关重要。利用生物信息学分析，我们的研究显示PP5在NSCLC组织中的表达明显高于正常组织。Kaplan-Meier生存分析还显示，PP5高表达与较差的总生存相关，特别是在接受放疗的患者中，这表明PP5在放射耐药中起作用。我们进一步证明PP5是palbociclib的关键靶点，不同于CDK4/6，影响NSCLC的放射敏感性。帕博西尼通过诱导持续DNA损伤和AMPK激活来增强放疗敏感性。随后的细胞事件是细胞凋亡而不是自噬。此外，联合治疗的增强疗效被AMPK抑制剂和PP5激活剂抵消，强调了这些途径在介导反应中的重要性。我们的研究结果提供了令人信服的证据，表明靶向PP5可以显著提高NSCLC放疗的治疗效果。这项研究为新的联合治疗策略提供了有价值的见解，突出了PP5作为克服放射耐药的新治疗靶点的潜力。

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引用次数: 0

RANKL regulates differentially breast cancer stem cell properties through its RANK and LGR4 receptors RANKL通过其RANK和LGR4受体调控乳腺癌干细胞的差异特性。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-02-01 DOI: 10.1016/j.bbamcr.2024.119888

Alejandro Ordaz-Ramos , Jorge Diaz-Blancas , Aketzalli Martínez-Cruz , Rosario Castro-Oropeza , Cecilia Zampedri , Damaris P Romero-Rodríguez , Mauricio Rodriguez-Dorantes , Jorge Melendez-Zajgla , Vilma Maldonado , Karla Vazquez-Santillan

Background

Breast cancer stem cells (BCSC) are a subpopulation responsible for cancer resistance and relapse. The receptor activator of nuclear factor kappa-Β ligand (RANKL) is a cytokine capable of activating RANK and LGR4 receptors. RANKL/RANK signaling maintains the self-renewal of BCSCs, however, the effect of RANKL via LGR4 remains unclear. Evidence from osteoclasts suggests that RANKL/LGR4 axis disrupts RANK signaling, leading to opposing cellular responses. Anti-RANKL inhibitors are potential agents for eradicating CSCs, but their effect on RANKL/LGR4 signal has not been demonstrated.

Objective

This project aimed to elucidate the role of RANKL in regulating stemness depending on the expression of its receptors.

Methods

We use in vitro and in vivo approaches to evaluate the effects of RANKL inhibition in stemness in low or high-LGR4 expressing cells. Furthermore, we analyze the effects of RANKL stimulation on the stemness of LGR4 or RANK overexpressing cells. Additionally, we evaluated the impact of RANKL/LGR4 signaling in the activity of Wnt/β-catenin and NF-κB signaling pathways.

Results

Our findings indicated that elevated RANKL expression is related to a favorable prognosis in patients with high LGR4 levels. Furthermore, RANKL inhibition decreased BCSC properties in LGR4-low cell lines, while it promoted migration in LGR4-high cells. Additionally, the RANKL/RANK axis activated NF-κB signaling and enhanced BCSCs in RANK-overexpressing cells. In contrast, in LGR4-overexpressing cells, RANKL failed to activate NF-κB but instead inhibited the Wnt/β-catenin pathway, leading to a reduction in BCSCs.

Conclusion

Our findings suggest that RANKL exerts different responses according to the expression of its receptors.

背景：乳腺癌干细胞（BCSC）是一个负责癌症抵抗和复发的亚群。核因子κ κ -Β配体受体激活因子（RANKL）是一种能够激活RANK和LGR4受体的细胞因子。RANKL/RANK信号维持BCSCs的自我更新，然而，RANKL通过LGR4的作用尚不清楚。来自破骨细胞的证据表明，RANKL/LGR4轴破坏RANK信号，导致相反的细胞反应。抗RANKL抑制剂是清除CSCs的潜在药物，但其对RANKL/LGR4信号的影响尚未得到证实。目的：本项目旨在阐明RANKL通过其受体的表达调控干细胞的作用。方法：采用体外和体内两种方法，对低表达或高表达lgr4的细胞进行RANKL抑制对干细胞的影响。此外，我们分析了RANKL刺激对LGR4或RANK过表达细胞的干性的影响。此外，我们评估了RANKL/LGR4信号通路对Wnt/β-catenin和NF-κB信号通路活性的影响。结果：我们的研究结果表明，在高LGR4水平的患者中，RANKL表达升高与良好的预后有关。此外，RANKL抑制降低了lgr4低的细胞系的BCSC特性，而促进了lgr4高的细胞的迁移。此外，RANKL/RANK轴激活NF-κB信号通路，并在RANK过表达的细胞中增强BCSCs。相比之下，在lgr4过表达的细胞中，RANKL不能激活NF-κB，而是抑制Wnt/β-catenin通路，导致BCSCs减少。结论：RANKL根据其受体的表达而产生不同的反应。

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引用次数: 0

GATA3: Orchestrating cellular fate through differentiation and proliferation GATA3：通过分化和增殖协调细胞命运。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-02-01 DOI: 10.1016/j.bbamcr.2024.119893

Rim Bacha , Shona Pedersen , Rana Ismail , Nouran Alwisi , Layla Al-Mansoori

Cell proliferation and differentiation are two fundamental biological processes that occur in biological systems, tightly regulated by various factors such as transcription factors (TFs). Zinc finger proteins are TFs responsible for maintaining the biological balance via coordinating development and functionality within the living cells. GATA binding protein 3 (GATA3), one of the zinc finger proteins, plays an essential role in driving differentiation and proliferation-related processes, thereby contributing to the regulation of the dynamism and productivity of living cells. By elucidating the complex interactions governed by GATA3, this underscores its significance in maintaining cellular homeostasis. Thus, the current review delves into the molecular pathways influenced by GATA3, highlighting its involvement in multiple developmental processes of various tissues and body sites, particularly in the hematopoietic system (T-cell differentiation), neural tissue differentiation, adipose tissue, as well as epithelial cell maturation.

细胞增殖和分化是发生在生物系统中的两个基本生物学过程，受到转录因子等多种因素的严格调控。锌指蛋白是通过协调活细胞内的发育和功能来维持生物平衡的tf。GATA结合蛋白3 （GATA binding protein 3, GATA3）是锌指蛋白之一，在细胞分化和增殖相关过程中发挥重要作用，对细胞的活力和生产力具有调节作用。通过阐明GATA3控制的复杂相互作用，这强调了它在维持细胞稳态中的重要性。因此，本综述深入探讨了受GATA3影响的分子途径，强调其参与多种组织和身体部位的多种发育过程，特别是造血系统（t细胞分化）、神经组织分化、脂肪组织以及上皮细胞成熟。

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引用次数: 0

Dedicator of cytokinesis protein 2 activates the epithelial–mesenchymal transition in renal fibrosis through the Rac1/PI3K/AKT pathway 细胞分裂献身蛋白2通过Rac1/PI3K/AKT通路激活肾纤维化的上皮-间质转化。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-02-01 DOI: 10.1016/j.bbamcr.2024.119894

Yuanyuan Jia , Jing Sun , Sha Chen , Yu Bian , Anni Jiang , Haihai Liang , Xuanyi Du

Renal fibrosis is the most important feature of the progression of chronic kidney disease (CKD), and epithelial–mesenchymal transition (EMT) plays an important role in renal fibrosis. Dedicator of cytokinesis protein 2 (Dock2) is involved in the immune system and the development of a variety of fibrotic diseases. However, its specific role in renal fibrosis remains unclear. Therefore, in this study, we investigated the role and mechanism of Dock2 in renal fibrosis. We constructed an in vivo mouse model of unilateral ureteral obstruction (UUO) and an in vitro model of recombinant human transforming growth factor-β1 (TGF-β1)-induced HK-2 cells. The function and regulatory mechanism of Dock2 were studied via Western blotting, qRT-PCR, immunohistochemistry and immunofluorescence. First, Dock2 was more highly expressed in the kidneys of UUO mice than in those of sham-operated mice. A reduction in Dock2 can improve pathological changes in the kidney tissue of UUO mice, reduce the deposition of the extracellular matrix (ECM), and alleviate EMT. Silencing Dock2 reduced the activation of both the Rac1 pathway and the PI3K/AKT pathway. TGF-β1 promoted Dock2 expression in HK-2 cells in vitro. A decrease in Dock2 can inhibit the expression of Fibronectin, Collagen I, α-SMA and Vimentin and increase the level of E-cadherin. Treatment of HK-2 cells with the Rac1 activator 8-CPT or the PI3K/AKT pathway activator YS-49 inhibited the above changes induced by siDock2, indicating that Dock2 activates EMT in renal fibrosis through the Rac1/PI3K/AKT pathway. Our data suggest that Dock2 may be a potential target for renal fibrosis treatment.

肾纤维化是慢性肾病（CKD）进展的最重要特征，上皮间质转化（epithelial-mesenchymal transition， EMT）在肾纤维化中起重要作用。细胞分裂献身蛋白2 （Dock2）参与免疫系统和多种纤维化疾病的发展。然而，其在肾纤维化中的具体作用尚不清楚。因此，在本研究中，我们研究了Dock2在肾纤维化中的作用和机制。建立小鼠单侧输尿管梗阻（UUO）体内模型和重组人转化生长因子-β1 （TGF-β1）诱导HK-2细胞体外模型。采用Western blotting、qRT-PCR、免疫组织化学和免疫荧光等方法研究Dock2的功能和调控机制。首先，Dock2在UUO小鼠肾脏中的表达高于假手术小鼠。Dock2的减少可以改善UUO小鼠肾组织的病理改变，减少细胞外基质（ECM）的沉积，减轻EMT。沉默Dock2降低了Rac1通路和PI3K/AKT通路的激活。TGF-β1在体外促进HK-2细胞Dock2的表达。Dock2降低可抑制纤维连接蛋白、I型胶原、α-SMA和Vimentin的表达，升高E-cadherin的水平。用Rac1激活剂8-CPT或PI3K/AKT通路激活剂YS-49处理HK-2细胞可抑制siDock2诱导的上述变化，表明Dock2通过Rac1/PI3K/AKT通路激活肾纤维化中的EMT。我们的数据表明Dock2可能是肾纤维化治疗的潜在靶点。

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引用次数: 0

Vasoactive intestinal peptide induces metabolic rewiring of human-derived cytotrophoblast cells to promote cell migration 血管活性肠肽诱导人源性细胞滋养层细胞代谢重组，促进细胞迁移。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-02-01 DOI: 10.1016/j.bbamcr.2024.119886

Fátima Merech, Brenda Lara, Daiana Rios, Daniel Paparini, Rosanna Ramhorst, Vanesa Hauk, Claudia Pérez Leirós , Daiana Vota

The placenta has an extraordinary metabolic rate with high oxygen consumption. Extravillous cytotrophoblast cells (EVT) metabolism and function are critical to sustain their invasive phenotype supporting fetal development. Deficient EVT function underlies pregnancy complications as preeclampsia (PE) and fetal growth restriction (FGR). The vasoactive intestinal peptide (VIP) promotes human cytotrophoblast cell migration and invasion through mTOR signaling pathways suggesting its crucial role during placentation. Here we explored fatty acid uptake as well as lipid and glucose metabolism in human-derived cytotrophoblast cell function upon VIP stimulation. We found that VIP induced long chain fatty acid (LCFAs) uptake along with the expression of FATP2 transporter, CPT1 fatty acid oxidation (FAO)-rate limiting step importer, and lipid droplet accumulation. VIP induced the expression of glucose 6-P-dehydrogenase, a rate-limiting enzyme of the pentose phosphate pathway (PPP) and pyruvate dehydrogenase complex enzyme DLAT E2, without altering lactate secretion. This metabolic rewiring of trophoblast cells induced by VIP takes place without compromising mitochondrial function or reactive oxygen species (ROS) production. Moreover, cytotrophoblast cell migration induced by VIP required the three glycolysis, oxidative phosphorylation (OXPHOS) and FAO pathways. Our results provide evidence supporting VIP as a metabolic regulatory peptide in cytotrophoblast cells sustaining proper placentation and fetal growth.

胎盘的代谢率非常高，耗氧量也很高。体外细胞滋养层细胞（EVT）的代谢和功能是维持其侵袭性表型支持胎儿发育的关键。EVT功能缺陷是妊娠并发症如先兆子痫（PE）和胎儿生长受限（FGR）的基础。血管活性肠肽（VIP）通过mTOR信号通路促进人细胞滋养层细胞迁移和侵袭，提示其在胎盘形成过程中起重要作用。本研究探讨了VIP刺激对人源性细胞滋养层细胞功能的脂肪酸摄取、脂质和糖代谢的影响。我们发现VIP诱导长链脂肪酸（LCFAs）摄取，同时表达FATP2转运蛋白、CPT1脂肪酸氧化（FAO）速率限制步骤入口蛋白和脂滴积累。VIP诱导葡萄糖6- p -脱氢酶（戊糖磷酸途径的限速酶）和丙酮酸脱氢酶复合物DLAT E2的表达，但不改变乳酸分泌。这种由VIP诱导的滋养细胞的代谢重新布线在不影响线粒体功能或活性氧（ROS）产生的情况下发生。此外，VIP诱导的细胞滋养层细胞迁移需要糖酵解、氧化磷酸化（OXPHOS）和FAO三种途径。我们的研究结果提供了支持VIP作为细胞滋养层细胞维持正常胎盘和胎儿生长的代谢调节肽的证据。

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引用次数: 0

A fluorescent protein C-terminal fusion knock-in is functional with TRPA1 but not TRPC5 荧光蛋白c端融合敲入对TRPA1起作用，但对TRPC5不起作用。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-02-01 DOI: 10.1016/j.bbamcr.2024.119887

Aaron Tragl , Alexandra Ptakova , Viktor Sinica , Rathej Meerupally , Christine König , Carolina Roza , Ivan Barvík , Viktorie Vlachova , Katharina Zimmermann

Objective

Transgenic mice with fluorescent protein (FP) reporters take full advantage of new in vivo imaging technologies. Therefore, we generated a TRPC5- and a TRPA1-reporter mouse based on FP C-terminal fusion, providing us with better alternatives for studying the physiology, interaction and coeffectors of these two TRP channels at the cellular and tissue level.

Methods

We generated transgenic constructs of the murine TRPC5- and TRPA1-gene with a 3*GGGGS linker and C-terminal fusion to mCherry and mTagBFP, respectively. We microinjected zygotes to generate reporter mice. Reporter mice were examined for visible fluorescence in trigeminal ganglia with two-photon microscopy, immunohistochemistry and calcium imaging.

Results

Both TRPC5-mCherry and TRPA1-mTagBFP knock-in mouse models were successful at the DNA and RNA level. However, at the protein level, TRPC5 resulted in no mCherry fluorescence. In contrast, sensory neurons derived from the TRPA1-reporter mice exhibited visible mTag-BFP fluorescence, although TRPA1 had apparently lost its ion channel function.

Conclusions

Creating transgenic mice with a TRP channel tagged at the C-terminus with a FP requires detailed investigation of the structural and functional consequences in a given cellular context and fine-tuning the design of specific constructs for a given TRP channel subtype. Different degrees of functional impairment of TRPA1 and TRPC5 constructs suggest a specific importance of the distal C-terminus for the regulation of these two channels in trigeminal neurons.

目的：荧光蛋白（FP）报告基因转基因小鼠充分利用新的体内成像技术。因此，我们基于FP c端融合构建了TRPC5-和trpa1报告小鼠，为我们在细胞和组织水平上研究这两个TRP通道的生理、相互作用和影响因子提供了更好的选择。方法：构建小鼠TRPC5-和trpa1基因的3*GGGGS连接体，分别与mCherry和mTagBFP进行c端融合。我们微注射受精卵来产生报告小鼠。采用双光子显微镜、免疫组织化学和钙显像检测三叉神经节可见荧光。结果：TRPC5-mCherry和TRPA1-mTagBFP敲入小鼠模型在DNA和RNA水平上均成功。然而，在蛋白水平上，TRPC5不产生mCherry荧光。相比之下，来自TRPA1报告小鼠的感觉神经元显示出可见的mTag-BFP荧光，尽管TRPA1明显失去了其离子通道功能。结论：在c端用FP标记TRP通道的转基因小鼠需要在给定的细胞环境中对结构和功能后果进行详细的研究，并对特定TRP通道亚型的特定结构进行微调设计。TRPA1和TRPC5结构的不同程度的功能损伤表明，远端c端对三叉神经元中这两个通道的调节具有特殊的重要性。

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引用次数: 0

Blockage of polycystin-2 alleviates myocardial ischemia/reperfusion injury by inhibiting autophagy through the Ca2+/Akt/Beclin 1 pathway 通过Ca2+/Akt/Beclin 1途径抑制自噬，阻断多囊卵巢素-2可减轻心肌缺血再灌注损伤。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-02-01 DOI: 10.1016/j.bbamcr.2024.119892

Xiao-dan Qin , Jian-feng Liang , Lin-yu Gan , Ke-shan Peng , Xue-hong Huang , Xiao-ting Li , Jin-li Chen , Wan Li , Lei Zhang , Jie Jian , Jun Lu

Autophagy is a well-conserved self-protection process that plays an important role in cardiovascular diseases. Excessive autophagy during myocardial ischemia/reperfusion injury (MIRI) induces calcium overload and the overactivation of an autophagic response, thereby aggravating cardiomyocyte damage. Polycystin-2 (PC2) is a Ca²⁺-permeable nonselective cation channel implicated in the regulation of autophagy. In the present study, autophagy was upregulated in myocardial ischemia/reperfusion in vivo and in vitro. PC2 knockdown using adeno-associated virus 9 particles containing Pkd2 short hairpin RNA infection markedly ameliorated MIRI, evidenced by reduced infarct size, diminished morphological changes, decreased cTnI levels, and improved cardiac function. Silencing PC2 reduced the autophagic flux in H9c2 cells. PC2 overexpression-mediated autophagic flux was inhibited by intracellular Ca²⁺ chelation with BAPTA-AM. Furthermore, PC2 ablation upregulated p-Akt (Ser473) and downregulated Beclin 1 in H/R. BAPTA-AM downregulated p-Akt(Ser473) and upregulated Beclin 1in PC2-overexpressing H9c2 cells. Moreover, the Akt inhibitor MK2206 abolished the BAPTA-AM-blunted PC2-dependent control of autophagy. Collectively, these results indicated that blockade of PC2 may be associated with the Ca²⁺/Akt/Beclin 1 signaling, thereby inhibiting excessive autophagy and serving as a potential strategy for mitigating MIRI.

自噬是一种保守的自我保护过程，在心血管疾病中起重要作用。心肌缺血/再灌注损伤（MIRI）时过度自噬诱导钙超载和自噬反应过度激活，从而加重心肌细胞损伤。多囊蛋白-2 （PC2）是一种参与自噬调节的Ca2+渗透性非选择性阳离子通道。在本研究中，自噬在体内和体外心肌缺血/再灌注中上调。使用含有Pkd2短发夹RNA感染的腺相关病毒9颗粒敲低PC2可显著改善MIRI，表现为梗死面积减小、形态学改变减少、cTnI水平降低和心功能改善。沉默PC2可降低H9c2细胞的自噬通量。细胞内Ca2+与BAPTA-AM螯合可抑制PC2过表达介导的自噬通量。此外，PC2消融可上调H/R中的p-Akt (Ser473)，下调Beclin 1。在pc2过表达的H9c2细胞中，BAPTA-AM下调p-Akt(Ser473)，上调Beclin 1。此外，Akt抑制剂MK2206消除了bapta - am钝化的pc2依赖性自噬控制。总之，这些结果表明，阻断PC2可能与Ca2+/Akt/Beclin 1信号传导有关，从而抑制过度自噬，并作为减轻MIRI的潜在策略。

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引用次数: 0

Fidgetin binds spastin to attenuate the microtubule-severing activity Fidgetin结合spastin减弱微管切断活性。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-02-01 DOI: 10.1016/j.bbamcr.2024.119890

Ying Sun , Skandha Ramakrishnan , Xiaona Lai , Ronghua Wu , Zhangji Dong , Liang Qiang , Mei Liu

Microtubule-severing enzymes such as spastin, katanin, and fidgetin, characterized by their AAA ATPase domains, are pivotal in modulating microtubule dynamics and behavior across various cellular processes. While spastin and katanin are recognized for their predominant and robust severing of stable microtubules, thereby enhancing microtubule turnover, fidgetin exhibits comparatively weaker severing activity and selectively targets labile microtubules. The interplay among these enzymes and their mutual regulatory mechanisms remains inadequately understood. In this study, we elucidate the functional interaction between spastin and fidgetin, focusing on their roles in microtubule severing and neurite outgrowth. Our findings demonstrate that fidgetin serves as a negative regulator of spastin's severing activity. Co-expression assays revealed that fidgetin significantly attenuates spastin's severing efficiency, as confirmed by fluorescence-based microtubule polymerization assays and quantitative imaging of microtubule dynamics. Co-immunoprecipitation and Förster Resonance Energy Transfer (FRET) analyses further established a direct interaction between fidgetin and spastin, suggesting that fidgetin modulates spastin's activity through direct binding, possibly contributing to forming the hetero-hexmeric ring for their severing activities. Functionally, spastin overexpression in neuronal cells enhances neurite outgrowth, an effect that is suppressed upon co-expression with fidgetin, indicating that fidgetin counterbalances spastin's activity to regulate neurite extension. Therefore, this study uncovers a previously unrecognized mechanism by which fidgetin modulates spastin's function, providing critical insights into the intricate regulation of microtubule severing. These findings have significant implications for therapeutic strategies targeting microtubule-severing activities, particularly in neurodevelopmental and neurodegenerative disorders where microtubule dysregulation is a hallmark.

微管切断酶，如spastin、katanin和fidgetin，以其AAA atp酶结构域为特征，在调节微管动力学和各种细胞过程中的行为中起关键作用。spastin和katanin主要切断稳定的微管，从而增强微管的周转，而fidgetin表现出相对较弱的切断活性，选择性地靶向不稳定的微管。这些酶之间的相互作用及其相互调节机制仍不充分了解。在这项研究中，我们阐明了痉挛蛋白和烦躁蛋白之间的功能相互作用，重点研究了它们在微管切断和神经突生长中的作用。我们的研究结果表明，烦躁素对痉挛素的切断活性起负调节作用。共表达实验显示，荧光微管聚合实验和微管动力学定量成像证实，fidgetin显著减弱了spastin的切断效率。共免疫沉淀和Förster共振能量转移（FRET）分析进一步建立了fidgetin和spastin之间的直接相互作用，表明fidgetin通过直接结合调节spastin的活性，可能有助于形成其切断活性的异六聚环。在功能上，神经细胞中痉挛蛋白的过表达增强了神经突的生长，这一作用在与烦躁蛋白共表达时被抑制，表明烦躁蛋白抵消了痉挛蛋白的活性来调节神经突的延伸。因此，这项研究揭示了一个以前未被认识到的机制，通过该机制，坐立不安素调节spastin的功能，为微管切断的复杂调节提供了重要的见解。这些发现对针对微管切断活动的治疗策略具有重要意义，特别是在微管失调是标志的神经发育和神经退行性疾病中。

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引用次数: 0

GALNT6, transcriptionally inhibited by KLF9, promotes osteosarcoma progression by increasing EFEMP1 expression via O-glycosylation modification 受 KLF9 转录抑制的 GALNT6 可通过 O 型糖基化修饰增加 EFEMP1 的表达，从而促进骨肉瘤的进展。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-02-01 DOI: 10.1016/j.bbamcr.2024.119879

Ziyuan Tong, Yuan Shen, Quan Yuan, Honghao Yu

Osteosarcoma (OS) is one of the deadliest malignancies in adolescents and its treatment status and prognosis remain unsatisfactory. N-acetylgalactosamine transferase 6 (GALNT6), one of the key enzymes regulating O-glycosylation, functions vary in different types of cancer. Currently, the function of GALNT6 in OS is unclear. Our results showed that GALNT6 was highly expressed in OS tissues, and the patients with higher GALNT6 expression exhibited a lower overall survival rate than patients with lower GALNT6 expression. We constructed the GALNT6-knockdown and GALNT6-overexpression vectors based on Tet-on system and packaged lentiviral particles to modulate GALNT6 expression. GALNT6 silencing impaired OC cell growth and metastasis both in vivo and vitro. Kruppel-like factor 9 (KLF9), a transcription factor known to suppress OS progression, was found to block GALNT6 transcription by binding to its promoter. Meanwhile, GALNT6 overexpression restored the effects caused by KLF9 upregulation. GALNT6 was known to affect protein stability by O-glycosylation regulation, thus the label-free proteomics combined with co-immunoprecipitation/mass-spectrum (MS) analysis were conducted to identify the potential mechanism of GALNT6 in promoting OS progression. EGF-containing fibulin extracellular matrix protein 1 (EFEMP1), contained several O-glycosylation sites and was upregulated in GALNT6 overexpressing cells (Log₂FC = 1.3195, p = 0.0160), attracted our attention. We demonstrated that GALNT6 interacted with EFEMP1 at protein level. The O-glycosylation of EFEMP1 was increased by GALNT6 overexpression, which slowed the degradation rate of EFEMP1. EFEMP1 knockdown reversed the effects of GALNT6 overexpression. Collectively, our observations demonstrate that KLF9/GALNT6/EFEMP1 may be a promising direction for OS treatment.

骨肉瘤（Osteosarcoma，OS）是青少年中最致命的恶性肿瘤之一，其治疗状况和预后仍不令人满意。N-乙酰半乳糖胺转移酶6（GALNT6）是调节O-糖基化的关键酶之一，在不同类型的癌症中功能各异。目前，GALNT6在OS中的功能尚不清楚。我们的研究结果表明，GALNT6在OS组织中高表达，GALNT6表达较高的患者总生存率低于GALNT6表达较低的患者。我们基于Tet-on系统构建了GALNT6-敲除和GALNT6-高表达载体，并包装慢病毒颗粒来调节GALNT6的表达。GALNT6沉默可抑制OC细胞在体内和体外的生长和转移。研究发现，Kruppel样因子9（KLF9）是一种已知能抑制OS进展的转录因子，它能通过与其启动子结合来阻断GALNT6的转录。同时，GALNT6的过表达可恢复KLF9上调所造成的影响。已知GALNT6通过O-糖基化调控影响蛋白质的稳定性，因此进行了无标记蛋白质组学结合共沉淀/质谱（MS）分析，以确定GALNT6促进OS进展的潜在机制。含EGF的纤维蛋白细胞外基质蛋白1（EFEMP1）含有多个O-糖基化位点，并在GALNT6过表达细胞中上调（Log2FC = 1.3195, p = 0.0160），这引起了我们的注意。我们证实了 GALNT6 与 EFEMP1 在蛋白质水平上的相互作用。GALNT6 的过表达增加了 EFEMP1 的 O 型糖基化，从而减缓了 EFEMP1 的降解速度。EFEMP1 的敲除逆转了 GALNT6 过表达的影响。总之，我们的观察结果表明，KLF9/GALNT6/EFEMP1可能是治疗OS的一个有前途的方向。

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引用次数: 0