Biochimica et biophysica acta. Molecular cell research最新文献_第7页

USP7 facilitates deubiquitination of LRRC42 in colorectal cancer to accelerate tumorigenesis and augment Wnt/β-catenin signaling USP7 在结直肠癌中促进 LRRC42 的去泛素化，从而加速肿瘤发生并增强 Wnt/β-catenin 信号传导。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2024-10-09 DOI: 10.1016/j.bbamcr.2024.119859

Yunze Li , Xin Sun , Zhe Huang

Colorectal cancer is a prevalent malignancy with an increasing incidence worldwide. Leucine-rich repeat-containing protein 42 (LRRC42) is known to be dysregulated in tumor tissues, yet its role in colorectal cancer remains largely unexplored. Herein, the function of LRRC42 in colorectal cancer was investigated using clinical samples, cellular experiments, animal models, and multiple omics techniques. The results demonstrated that LRRC42 was highly expressed in colorectal cancer tissues and was associated with poor clinical outcomes. Silencing LRRC42 suppressed cell proliferation, induced G0/G1 phase arrest, and promoted apoptosis by reducing Bcl2 expression while elevating the expression of Bax, cleaved PARP and cleaved caspase 3. Conversely, LRRC42 overexpression exhibited the opposite effects. Consistent findings were observed in vivo. Additionally, ubiquitin specific peptidase 7 was identified as a potential LRRC42-interacting protein through immunoprecipitation-mass spectrometry, with ubiquitin specific peptidase 7 stabilizing LRRC42 expression by promoting its deubiquitination. Notably, LRRC42 overexpression partially reversed the effects of ubiquitin specific peptidase 7 silencing on tumor cell proliferation and apoptosis. mRNA sequencing analysis revealed that differentially expressed genes in LRRC42 overexpressing cells were linked to Wnt signaling pathway, suggesting that LRRC42 overexpression may activate this pathway. Furthermore, LRRC42 was proved to elevate the levels of ki67, cyclin D1 and WNT3, while reducing the level of p-β-catenin. These findings suggest that LRRC42 perhaps serve as a potential oncogenic factor in colorectal cancer, regulated by ubiquitin specific peptidase 7 and capable of activating Wnt/β-catenin signaling pathway.

结直肠癌是一种常见的恶性肿瘤，在全球的发病率不断上升。众所周知，富亮氨酸重复序列蛋白 42（LRRC42）在肿瘤组织中调控失调，但其在结直肠癌中的作用仍未得到充分探究。在此，研究人员利用临床样本、细胞实验、动物模型和多种全息技术研究了LRRC42在结直肠癌中的功能。结果表明，LRRC42在结直肠癌组织中高表达，并与不良临床预后相关。沉默 LRRC42 可抑制细胞增殖，诱导 G0/G1 期停滞，并通过降低 Bcl2 表达，同时提高 Bax、裂解 PARP 和裂解 Caspase 3 的表达来促进细胞凋亡。相反，LRRC42 过表达则表现出相反的效果。在体内也观察到了一致的结果。此外，通过免疫沉淀-质谱分析，泛素特异性肽酶 7 被确定为潜在的 LRRC42 相互作用蛋白，泛素特异性肽酶 7 通过促进 LRRC42 的去泛素化来稳定其表达。mRNA测序分析发现，LRRC42过表达细胞中的差异表达基因与Wnt信号通路有关，表明LRRC42的过表达可能激活了这一通路。此外，LRRC42 还能提高 ki67、细胞周期蛋白 D1 和 WNT3 的水平，同时降低 p-β-catenin 的水平。这些研究结果表明，LRRC42 可能是结直肠癌的潜在致癌因子，它受泛素特异性肽酶 7 的调控，能够激活 Wnt/β-catenin 信号通路。

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引用次数: 0

Atrial natriuretic peptide (ANP) modulates stress-induced autophagy in endothelial cells 心房利钠肽（ANP）可调节内皮细胞中由压力诱导的自噬。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2024-10-09 DOI: 10.1016/j.bbamcr.2024.119860

Maurizio Forte , Simona Marchitti , Flavio di Nonno , Donatella Pietrangelo , Rosita Stanzione , Maria Cotugno , Luca D'Ambrosio , Alessandra D'Amico , Vittoria Cammisotto , Gianmarco Sarto , Erica Rocco , Beatrice Simeone , Sonia Schiavon , Daniele Vecchio , Roberto Carnevale , Salvatore Raffa , Giacomo Frati , Massimo Volpe , Sebastiano Sciarretta , Speranza Rubattu

Atrial natriuretic peptide (ANP), a cardiac hormone involved in the regulation of water/sodium balance and blood pressure, is also secreted by endothelial cells, where it exerts protective effects in response to stress. Autophagy is an intracellular self-renewal process involved in the degradation of dysfunctional cytoplasmic elements. ANP was recently reported to act as an extracellular regulator of cardiac autophagy. However, its role in the regulation of endothelial autophagy has never been investigated. Here, we tested the effects of ANP in the regulation of autophagy in human umbilical vein endothelial cells (HUVECs). We found that ANP rapidly increases autophagy and autophagic flux at physiological concentrations through its predominant pathway, mediated by natriuretic peptide receptor type A (NPR-A) and protein kinase G (PKG). We further observed that ANP is rapidly secreted by HUVEC under stress conditions, where it mediates stress-induced autophagy through autocrine and paracrine mechanisms. Finally, we found that the protective effects of ANP in response to high-salt loading or tumor necrosis factor (TNF)-α are blunted by concomitant inhibition of autophagy. Overall, our results suggest that ANP acts as an endogenous autophagy activator in endothelial cells. The autophagy mechanism mediates the protective endothelial effects exerted by ANP.

心房利钠肽（ANP）是一种参与调节水/钠平衡和血压的心脏激素，它也由内皮细胞分泌，在应对压力时发挥保护作用。自噬是一种细胞内自我更新过程，参与降解功能失调的细胞质元素。最近有报道称，ANP 是心脏自噬的细胞外调节因子。然而，它在调节内皮自噬中的作用却从未被研究过。在这里，我们测试了 ANP 在调节人脐静脉内皮细胞（HUVECs）自噬中的作用。我们发现，在生理浓度下，ANP 通过其主要途径迅速增加自噬和自噬通量，该途径由钠肽受体 A 型（NPR-A）和蛋白激酶 G（PKG）介导。我们进一步观察到，HUVEC 在应激条件下会快速分泌 ANP，ANP 通过自分泌和旁分泌机制介导应激诱导的自噬。最后，我们发现 ANP 对高盐负荷或肿瘤坏死因子（TNF）-α 的保护作用会因同时抑制自噬而减弱。总之，我们的研究结果表明，ANP 是内皮细胞中的内源性自噬激活剂。自噬机制介导了 ANP 对内皮细胞的保护作用。

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引用次数: 0

Structural analysis of microtubule binding by minus-end targeting protein Spiral2 负端靶向蛋白 Spiral2 与微管结合的结构分析

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2024-10-04 DOI: 10.1016/j.bbamcr.2024.119858

Marina Ohno , Yuuki Higuchi , Kazune Yamai , Sotaro Fuchigami , Takema Sasaki , Yoshihisa Oda , Ikuko Hayashi

Microtubules (MTs) are dynamic cytoskeletal polymers that play a critical role in determining cell polarity and shape. In plant cells, acentrosomal MTs are localized on the cell surface and are referred to as cortical MTs. Cortical MTs nucleate in the cell cortex and detach from nucleation sites. The released MT filaments perform treadmilling, with the plus-ends of MTs polymerizing and the minus-ends depolymerizing. Minus-end targeting proteins, -TIPs, include Spiral2, which regulates the minus-end dynamics of acentrosomal MTs. Spiral2 accumulates autonomously at MT minus-ends and inhibits filament shrinkage, but the mechanism by which Spiral2 specifically recognizes minus-ends of MTs remains unknown. Here we describe the crystal structure of Spiral2's N-terminal MT-binding domain. The structural properties of this domain resemble those of the HEAT repeat structure of the tumor overexpressed gene (TOG) domain, but the number of HEAT repeats is different and the conformation is highly arched. Gel filtration and co-sedimentation analyses demonstrate that the domain binds preferentially to MT filaments rather than the tubulin dimer, and that the tubulin-binding mode of Spiral2 via the basic surface is similar to that of the TOG domain. We constructed an in silico model of the Spiral2-tubulin complex to identify residues that potentially recognize tubulin. Mutational analysis revealed that the key residues inferred in the model are involved in microtubule recognition, and provide insight into the mechanism by which end-targeting proteins stabilize MT ends.

微管（MT）是一种动态细胞骨架聚合物，在决定细胞极性和形状方面起着至关重要的作用。在植物细胞中，顶体 MT 定位于细胞表面，被称为皮层 MT。皮层 MT 在细胞皮层成核，并从成核点分离。释放出来的MT丝进行踩踏运动，MT的正端聚合，负端解聚。负端靶向蛋白（-TIPs）包括螺旋2（Spiral2），它能调节顶体MT的负端动态。Spiral2 在 MT 负端自主聚集并抑制纤维收缩，但 Spiral2 特异性识别 MT 负端的机制仍不清楚。在这里，我们描述了 Spiral2 N 端 MT 结合结构域的晶体结构。该结构域的结构特性与肿瘤过表达基因（TOG）结构域的 HEAT 重复结构相似，但 HEAT 重复的数量不同，而且构象呈高度弧形。凝胶过滤和共沉淀分析表明，该结构域优先与 MT 细丝结合，而不是与微管蛋白二聚体结合，而且 Spiral2 通过基本表面与微管蛋白结合的模式与 TOG 结构域类似。我们构建了一个Spiral2-微管蛋白复合物的硅学模型，以确定可能识别微管蛋白的残基。突变分析表明，模型中推断出的关键残基参与了微管识别，并为末端靶向蛋白稳定MT末端的机制提供了启示。

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引用次数: 0

CISD2 counteracts the inhibition of ER-mitochondrial calcium transfer by anti-apoptotic BCL-2 CISD2 可抵消抗凋亡 BCL-2 对 ER 线粒体钙传递的抑制。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2024-10-04 DOI: 10.1016/j.bbamcr.2024.119857

Jens Loncke , Ian de Ridder , Justin Kale , Larry Wagner , Allen Kaasik , Jan B. Parys , Martijn Kerkhofs , David W. Andrews , David Yule , Tim Vervliet , Geert Bultynck

CISD2, a 2Fe2S cluster domain-containing protein, is implicated in Wolfram syndrome type 2, longevity and cancer. CISD2 is part of a ternary complex with IP₃ receptors (IP₃Rs) and anti-apoptotic BCL-2 proteins and enhances BCL-2's anti-autophagic function. Here, we examined how CISD2 impacted the function of BCL-2 in apoptosis and in controlling IP₃R-mediated Ca²⁺ signaling. Using purified proteins, we found a direct interaction between the cytosolic region of CISD2 and BCL-2's BH4 domain with a submicromolar affinity. At the functional level, the cytosolic region of CISD2, as a purified protein, did not affect the ability of BCL-2 to inhibit BAX-pore formation. In a cellular context, loss of CISD2 did not impede the suppression of apoptosis by BCL-2. Also, in Ca²⁺-signaling assays, absence of CISD2 did not affect the inhibition of IP₃R-mediated Ca²⁺ release by BCL-2. Combined, these experiments indicate that CISD2 is not essential for BCL-2 function in apoptosis and cytosolic Ca²⁺ signaling. Instead, CISD2 overexpression enhanced BCL-2-mediated suppression of cytosolic IP₃R-mediated Ca²⁺ release. However, consistent with the presence of CISD2 and BCL-2 at mitochondria-associated ER membranes (MAMs), the most striking effect was observed at the level of ER-mitochondrial Ca²⁺ transfer. While BCL-2 overexpression inhibited ER-mitochondrial Ca²⁺ transfer, overexpression of CISD2 together with BCL-2 abrogated the effect of BCL-2. The underlying mechanism is linked to ER-mitochondrial contact sites, since BCL-2 reduced ER-mitochondrial contact sites while co-expression of CISD2 together with BCL-2 abolished this effect. These findings reveal a unique interplay between BCL-2 and CISD2 at Ca²⁺-signaling nanodomains between ER and mitochondria.

CISD2是一种含2Fe2S簇结构域的蛋白质，与沃尔夫拉姆综合征2型、长寿和癌症有关。CISD2是IP3受体（IP3Rs）和抗凋亡BCL-2蛋白三元复合物的一部分，能增强BCL-2的抗自噬功能。在此，我们研究了 CISD2 如何影响 BCL-2 在细胞凋亡和控制 IP3R 介导的 Ca2+ 信号转导中的功能。通过纯化蛋白，我们发现 CISD2 的胞浆区与 BCL-2 的 BH4 结构域之间存在直接相互作用，亲和力达到亚摩尔级。在功能层面上，CISD2的胞浆区作为一种纯化蛋白，并不影响BCL-2抑制BAX孔形成的能力。在细胞环境中，CISD2的缺失并不妨碍BCL-2抑制细胞凋亡。此外，在 Ca2+ 信号传导实验中，CISD2 的缺失并不影响 BCL-2 对 IP3R 介导的 Ca2+ 释放的抑制作用。这些实验综合表明，CISD2 对于 BCL-2 在细胞凋亡和细胞膜 Ca2+ 信号转导中的功能并不是必不可少的。相反，CISD2 的过表达增强了 BCL-2 介导的对细胞膜 IP3R 介导的 Ca2+ 释放的抑制。然而，与 CISD2 和 BCL-2 存在于线粒体相关的 ER 膜（MAMs）一致，在 ER - 线粒体 Ca2+ 转移水平观察到了最显著的影响。BCL-2的过表达抑制了ER-线粒体Ca2+的转移，而CISD2与BCL-2一起过表达则削弱了BCL-2的作用。其潜在机制与ER-线粒体接触位点有关，因为BCL-2减少了ER-线粒体接触位点，而CISD2与BCL-2共同表达则消除了这种效应。这些发现揭示了 BCL-2 和 CISD2 在 ER 和线粒体之间的 Ca2+ 信号纳米域的独特相互作用。

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引用次数: 0

Effects of high-fat diet on folic acid-induced kidney injury in mice 高脂饮食对叶酸诱导的小鼠肾损伤的影响

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2024-09-30 DOI: 10.1016/j.bbamcr.2024.119856

Doyeon Kim , Minjung Son , Sugyeong Ha , Jeongwon Kim , Mi-Jeong Kim , Jian Yoo , Byeong Moo Kim , Hae Young Chung , Haeseung Lee , Donghwan Kim , Sangok Kim , Ki Wung Chung

Obesity is recognized as a significant contributor to the onset of kidney disease. However, the key processes involved in the development of kidney disease in obese individuals are not well understood. Here, we investigated the effects of high-fat diet (HFD)-induced obesity on folic acid (FA)-induced kidney injury in mice. Mice were fed an HFD for 12 weeks to induce obesity, followed by an additional intraperitoneal injection of FA. The results showed that mice fed HFD developed higher levels of kidney damage than those in the chow group. In contrast, mice exposed to both HFD and FA showed less fibrosis and inflammatory responses compared to the FA only treated group. Furthermore, the HFD with FA group exhibited elevated lipid accumulation in the kidney and reduced expression of mitochondrial proteins compared to the FA-treated group. Under in vitro experimental conditions, we found that lipid accumulation induced by oleic acid treatment reduced inflammatory and fibrotic responses in both renal tubules and fibroblasts. Finally, RNA sequencing analysis revealed that the inflammasome and pyroptosis signaling pathways were significantly increased in the HFD group with FA injection. In summary, these findings suggest that obesity increases renal injury due to a lack of appropriate inflammatory, fibrotic, and metabolic responses and the activation of the inflammasome and pyroptosis signaling pathways.

肥胖被认为是导致肾病发病的一个重要因素。然而，肥胖者肾脏疾病发生的关键过程尚不十分清楚。在此，我们研究了高脂饮食（HFD）诱导的肥胖对叶酸（FA）诱导的小鼠肾损伤的影响。给小鼠喂食高脂饮食 12 周以诱导肥胖，然后再腹腔注射叶酸。结果显示，喂食高纤维食物的小鼠比饲料组的小鼠出现更严重的肾损伤。相比之下，同时摄入 HFD 和 FA 的小鼠与只摄入 FA 的组相比，纤维化和炎症反应较轻。此外，与 FA 处理组相比，HFD 和 FA 组的肾脏脂质积累增加，线粒体蛋白表达减少。在体外实验条件下，我们发现油酸处理诱导的脂质积累减轻了肾小管和成纤维细胞的炎症和纤维化反应。最后，RNA 测序分析表明，注射 FA 的高脂血症组中，炎性体和裂解酶信号通路显著增加。总之，这些研究结果表明，由于缺乏适当的炎症、纤维化和新陈代谢反应以及炎性体和裂解酶信号通路的激活，肥胖会加重肾损伤。

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引用次数: 0

NLRX1 attenuates endoplasmic reticulum stress via STING in cardiac hypertrophy NLRX1 通过 STING 减轻心肌肥厚中的内质网应激反应

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2024-09-30 DOI: 10.1016/j.bbamcr.2024.119852

Keying Mi , Xiaoyan Wang , Chao Ma , Yinghua Tan , Gang Zhao , Xinran Cao , Haitao Yuan

Endoplasmic reticulum stress-induced cell apoptosis is a pivotal mechanism underlying the progression of cardiac hypertrophy. NLRX1, a member of the NOD-like receptor family, modulates various cellular processes, including STING, NF-κB, MAPK pathways, reactive oxygen species production, essential metabolic pathways, autophagy and cell death. Emerging evidence suggests that NLRX1 may offer protection against diverse cardiac diseases. However, the impacts and mechanisms of NLRX1 on endoplasmic reticulum stress in cardiac hypertrophy remains largely unexplored. In our study, we observed that the NLRX1 and phosphorylated STING (p-STING) were highly expressed in both hypertrophic mouse heart and cellular model of cardiac hypertrophy. Whereas over-expression of NLRX1 mitigated the expression levels of p-STING, as well as the endoplasmic reticulum stress markers, including transcription activating factor 4 (ATF4), C/EBP homologous protein (CHOP) and the ratios of phosphorylated PERK to PERK, phosphorylated IRE1 to IRE1 and phosphorylated eIF2α to eIF2α in an Angiotensin II (Ang II)-induced cellular model of cardiac hypertrophy. Importantly, the protective effects of NLRX1 were attenuated upon pretreatment with the STING agonist, DMXAA. Our findings provide the evidence that NLRX1 attenuates the PERK-eIF2α-ATF4-CHOP axis of endoplasmic reticulum stress response via inhibition of p-STING in Ang II-treated cardiomyocytes, thereby ameliorating the development of cardiac hypertrophy.

内质网应激诱导的细胞凋亡是心肌肥厚进展的关键机制。NLRX1 是 NOD 样受体家族的成员，它能调节各种细胞过程，包括 STING、NF-κB、MAPK 途径、活性氧生成、基本代谢途径、自噬和细胞死亡。新的证据表明，NLRX1 可预防多种心脏疾病。然而，NLRX1 对心脏肥大中内质网应激的影响和机制在很大程度上仍未得到探索。在我们的研究中，我们观察到 NLRX1 和磷酸化 STING（p-STING）在肥厚小鼠心脏和心脏肥厚细胞模型中均高表达。而在血管紧张素 II（Ang II）诱导的心脏肥大细胞模型中，过度表达 NLRX1 可减轻 p-STING 的表达水平，以及内质网应激标志物，包括转录激活因子 4（ATF4）、C/EBP 同源蛋白（CHOP）和磷酸化 PERK 与 PERK、磷酸化 IRE1 与 IRE1 和磷酸化 eIF2α 与 eIF2α 的比率。重要的是，在 STING 激动剂 DMXAA 的预处理下，NLRX1 的保护作用减弱。我们的研究结果提供了证据，证明 NLRX1 通过抑制 Ang II 处理的心肌细胞中的 p-STING 来减弱内质网应激反应的 PERK-eIF2α-ATF4-CHOP 轴，从而改善心脏肥大的发展。

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引用次数: 0

AGR2 knockdown induces ER stress and mitochondria fission to facilitate pancreatic cancer cell death AGR2 基因敲除可诱导ER应激和线粒体分裂，从而促进胰腺癌细胞死亡。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2024-09-30 DOI: 10.1016/j.bbamcr.2024.119854

Philip Salu , Daniel Tuvin , Katie M. Reindl

Anterior gradient 2 (AGR2) is often overexpressed in many human cancers, including pancreatic ductal adenocarcinoma (PDAC). Elevated AGR2 expression is known to play a critical role in tumor development, progression, and metastasis and positively correlates with poor patient survival. However, the relationship between AGR2 expression and tumor growth is not fully understood. Our study aims to investigate the impact of AGR2 knockdown on the survival of two pancreatic cancer cell lines, HPAF–II and PANC–1, that exhibit high AGR2 expression. This study revealed that the knockdown of AGR2 expression through an inducible shRNA-mediated approach reduced the proliferative ability and colony-forming potential of PDAC cells compared to scramble controls. Significantly, knocking down AGR2 led to the inhibition of multiple protein biosynthesis pathways and induced ER stress through unfolded protein response (UPR) activation. AGR2 knockdown induced ER stress and increased mitochondrial fission, while mitochondrial fusion remained unaffected. Ultimately, apoptotic cell death was heightened in AGR2 knockdown PDAC cells compared to the controls. Overall, these data reveal a new axis involving AGR2-ER stress-associated mitochondrial fission that could be targeted to improve PDAC patient outcomes.

前梯度 2（AGR2）经常在包括胰腺导管腺癌（PDAC）在内的许多人类癌症中过度表达。众所周知，AGR2 的高表达在肿瘤发生、发展和转移过程中起着至关重要的作用，并与患者生存率低呈正相关。然而，AGR2 表达与肿瘤生长之间的关系尚未完全明了。我们的研究旨在探讨 AGR2 基因敲除对两种高表达 AGR2 的胰腺癌细胞系 HPAF-II 和 PANC-1 生存的影响。研究发现，通过诱导性 shRNA 介导的方法敲除 AGR2 表达，与混杂对照组相比，可降低 PDAC 细胞的增殖能力和集落形成潜能。值得注意的是，敲除 AGR2 会抑制多种蛋白质的生物合成途径，并通过激活未折叠蛋白反应（UPR）诱导 ER 应激。敲除 AGR2 会诱发 ER 应激并增加线粒体裂变，而线粒体融合则不受影响。最终，与对照组相比，AGR2 基因敲除的 PDAC 细胞凋亡增加。总之，这些数据揭示了一个涉及 AGR2-ER应激相关线粒体裂变的新轴，可以针对该轴改善PDAC患者的预后。

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引用次数: 0

An artificial peptide inhibits autophagy through calcineurin-TFEB pathway 人工肽通过钙神经蛋白-TFEB途径抑制自噬。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2024-09-30 DOI: 10.1016/j.bbamcr.2024.119853

Yumeng Yang , Yanan Li , Hanxiao Shang , Yueyang Liu , Wenying Li , Limin Chen , Na Cheng , Yuchen Zhang , Nan Zhang , Yanxia Yin , Li Tong , Zhimei Li , Jingyu Yang , Jing Luo

We previously reported that a bioactive peptide (pep3) can potently inhibit the enzyme activity of purified calcineurin (CN). In this paper, we further demonstrate that transfected pep3 can strongly inhibit CN enzyme activity in HEK293 cells. Transcription factor EB (TFEB) plays an important role in the autophagy-lysosome pathway (ALP) as one of the substrates of CN, so we study the effect of pep3 on the CN-TFEB-ALP pathway. Pep3 can significantly inhibit the mRNA levels of the TFEB downstream genes and the expression of the autophagy-associated proteins, and autophagy flux in HEK293 cells. We also validated the inhibitory effect of pep3 on autophagy in mice. These findings may provide a new idea for discovering more CN inhibitors and autophagy inhibitory drugs.

我们以前曾报道过一种生物活性肽（pep3）能有效抑制纯化的钙调磷酸酶（CN）的酶活性。在本文中，我们进一步证明了转染的 pep3 能强烈抑制 HEK293 细胞中 CN 的酶活性。转录因子 EB（TFEB）作为 CN 的底物之一，在自噬-溶酶体通路（ALP）中发挥着重要作用，因此我们研究了 pep3 对 CN-TFEB-ALP 通路的影响。Pep3能明显抑制TFEB下游基因的mRNA水平、自噬相关蛋白的表达以及HEK293细胞的自噬通量。我们还在小鼠体内验证了 pep3 对自噬的抑制作用。这些发现为发现更多的CN抑制剂和自噬抑制药物提供了新思路。

引用次数: 0

TRIM40 interacts with ROCK1 directly and inhibits colorectal cancer cell proliferation through the c-Myc/p21 axis TRIM40 与 ROCK1 直接相互作用，通过 c-Myc/p21 轴抑制结直肠癌细胞增殖。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2024-09-30 DOI: 10.1016/j.bbamcr.2024.119855

Fangyu Hu , Lingling Zhao , Junyu Wang , Xiaoying Li , Zixuan Xue , Yimeng Ma , Minghui Zheng , Chenglin Chen , Meiting Tong , Xiaohuan Guo , Hongyan Li , Honglei Jin , Qipeng Xie , Xiaodong Zhang , Chuanshu Huang , Haishan Huang

Background

Colorectal cancer (CRC) is the most common malignancy of the digestive tract, and to date, morbidity and mortality rates remain high. While existing therapeutic methods have achieved certain effective outcomes, there are still many problems in treating this disease. Therefore, it is still urgent to constantly find new therapeutic targets in CRC that could lead to new therapeutics.

Methods

Immunohistochemistry, Real-time PCR and Western Blot were employed to measure mRNA and protein levels of the target protein, respectively. The proliferation ability of CRC cells was evaluated using ATP assay, Soft agar assay, and nude mouse subcutaneous tumorigenesis assay. Protein Degradation Assay was conducted to determine protein degradation rate, while Ubiquitination assay was used to assess the ubiquitination modification level of target proteins. Immunoprecipitation assay was used to study protein interactions, and pull-down assay was employed to investigate direct interactions between proteins.

Results

TRIM40 was significantly down-regulated in CRC tissues, with its expression levels positively correlating with disease prognosis. Using both in vitro and in vivo approaches, it was demonstrated that TRIM40 could significantly inhibit the proliferation of CRC cells. Molecular mechanism studies showed that TRIM40 directly binds to and ubiquitinates ROCK1 protein, accelerating its degradation and subsequently reducing the stability of c-Myc protein. This cascade of events results in the release of transcriptional inhibition of p21 by c-Myc, leading to increased p21 expression and G0/G1 phase arrest in CRC cells.

Conclusion

This research suggests that TRIM40 could be a valuable therapeutic target for the treatment of CRC.

背景：大肠癌（CRC）是消化道最常见的恶性肿瘤，迄今为止，发病率和死亡率仍然居高不下。虽然现有的治疗方法取得了一定的疗效，但在治疗该疾病方面仍存在许多问题。因此，不断寻找 CRC 的新治疗靶点，从而开发出新的治疗方法，仍是当务之急：方法：采用免疫组化、Real-time PCR和Western Blot技术分别检测靶蛋白的mRNA和蛋白水平。采用 ATP 试验、软琼脂试验和裸鼠皮下肿瘤发生试验评估 CRC 细胞的增殖能力。蛋白质降解试验用于测定蛋白质的降解率，泛素化试验用于评估目标蛋白质的泛素化修饰水平。免疫沉淀试验用于研究蛋白质之间的相互作用，而牵引试验则用于研究蛋白质之间的直接相互作用：结果：TRIM40在CRC组织中明显下调，其表达水平与疾病预后呈正相关。体外和体内研究表明，TRIM40能明显抑制CRC细胞的增殖。分子机制研究表明，TRIM40 可直接与 ROCK1 蛋白结合并泛素化，加速其降解，进而降低 c-Myc 蛋白的稳定性。这一系列事件导致c-Myc释放对p21的转录抑制，导致p21表达增加，CRC细胞G0/G1期停滞：这项研究表明，TRIM40 可能是治疗 CRC 的一个有价值的治疗靶点。

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引用次数: 0

Mitochondrial dynamics and sex-specific responses in the developing rat hippocampus: Effect of perinatal asphyxia and mesenchymal stem cell Secretome treatment 发育中大鼠海马的线粒体动态和性别特异性反应：围产期窒息和间充质干细胞Secretome处理的影响

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2024-09-25 DOI: 10.1016/j.bbamcr.2024.119851

M. Zamorano-Cataldo , I. Vega-Vásquez , C. García-Navarrete , J. Toledo , D. Bustamante , F. Ezquer , F.A. Urra , N. Farfán-Troncoso , M. Herrera-Marschitz , P. Morales

Aims

Perinatal asphyxia is one of the major causes of neonatal death at birth. Survivors can progress but often suffer from long-term sequelae. We aim to determine the effects of perinatal asphyxia on mitochondrial dynamics and whether mesenchymal stem cell secretome (MSC-S) treatment can alleviate the deleterious effects.

Materials and methods

Animals were subjected to 21 min of asphyxia at the time of delivery. MSC-S or vehicle was intranasally administered 2 h post-delivery. Mitochondrial mass (D-loop, qPCR), mitochondrial dynamics proteins (Drp1, Fis1 and OPA1, Western blot), mitochondrial dynamics (TOMM20, Immunofluorescence), as well as mitochondrial membrane potential (ΔΨ_m) (Safranin O) were evaluated at P1 and P7 in the hippocampus.

Key findings

Perinatal asphyxia increased levels of mitochondrial dynamics proteins Drp1 and S-OPA1 at P1 and Fis1 at P7. Mitochondrial density and mass were decreased at P1. Perinatal asphyxia induced sex-specific differences, with increased L-OPA1 in females at P7 and increased mitochondria circularity. In males, asphyxia-exposed animals exhibited a reduced ΔΨ_m at P7. MSC-S treatment normalised levels of mitochondrial dynamics proteins involved in fission.

Significance

This study provides novel insights into the effects of perinatal asphyxia on mitochondrial dynamics in the developing brain and on the therapeutic opportunities provided by mesenchymal stem cell secretome treatment. It also highlights on the relevance of considering sex as a biological variable in perinatal brain injury and therapy development. These findings contribute to the development of targeted, personalised therapies for infants affected by perinatal asphyxia.

目的：围产期窒息是新生儿出生时死亡的主要原因之一。存活者可以继续发育，但往往会留下长期后遗症。我们旨在确定围产期窒息对线粒体动力学的影响，以及间充质干细胞分泌物（MSC-S）治疗是否能减轻其有害影响：动物在分娩时窒息21分钟。材料与方法：动物在分娩时窒息 21 分钟，分娩后 2 小时鼻内注射间充质干细胞-S 或药物。在海马P1和P7时评估线粒体质量（D-loop，qPCR）、线粒体动力学蛋白（Drp1、Fis1和OPA1，Western印迹）、线粒体动力学（TOMM20，免疫荧光）以及线粒体膜电位（ΔΨm）（Safranin O）：主要研究结果：围产期窒息会增加线粒体动力学蛋白 Drp1 和 S-OPA1 的水平（P1）和 Fis1 的水平（P7）。线粒体密度和质量在P1时下降。围产期窒息引起了性别差异，雌性在P7时L-OPA1增加，线粒体循环性增加。在雄性动物中，窒息暴露的动物在P7时表现出ΔΨm减少。MSC-S处理可使参与裂变的线粒体动力学蛋白水平恢复正常：这项研究为围产期窒息对发育中大脑线粒体动力学的影响以及间充质干细胞分泌组治疗提供了新的见解。研究还强调了将性别视为围产期脑损伤和治疗发展中的一个生物变量的相关性。这些发现有助于为受围产期窒息影响的婴儿开发有针对性的个性化疗法。

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