Experimental cell research最新文献_第2页

SP1-mediated transcriptional repression of SFRP5 is correlated with cardiac fibroblast activation and atrial myocyte apoptosis in the development of atrial fibrillation SP1 介导的 SFRP5 转录抑制与心房颤动发生过程中心脏成纤维细胞活化和心房肌细胞凋亡有关。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2024-11-12 DOI: 10.1016/j.yexcr.2024.114326

Yanyan Sun, Zhenzhen Hu, Jie Han, Gang Li

Secreted frizzled related protein 5 (SFRP5) is a recognized cardioprotective protein with diminished expression in atrial fibrillation (AF). This study investigates SFRP5's function in AF-related cardiac fibrosis and cardiomyocyte apoptosis, exploring the underlying dysregulation causes. Utilizing C57BL/6 mice, mouse cardiac fibroblasts (CFs), and HC-1 mouse atrial myocyte cell line, AF models were induced by angiotensin Ⅱ (Ang Ⅱ). SFRP5 levels were consistently decreased in plasma samples from clinical patients, modeled mice, and CF culture supernatants. Treatment with recombinant SFRP5 restored its levels, mitigating Ang Ⅱ-induced AF in mice and ameliorating atrial tissue fibrosis and oxidative stress. In vitro, SFRP5 recombinant protein suppressed CF activation and fibrosis-related markers. The study identified Sp1 transcription factor (SP1) binding to the SFRP5 promoter, causing transcriptional repression. SP1 knockdown reinstated SFRP5 levels in mice and CFs, thus suppressing fibrosis. Additionally, SP1 knockdown attenuated Ang Ⅱ-induced apoptosis in HC-1 cells, but this effect was counteracted by concurrent SFRP5 knockdown. In conclusion, this investigation underscores that SP1 mediates SFRP5 loss during AF by transcriptional repression, contributing to fibrosis and myocyte apoptosis. These findings illuminate potential therapeutic interventions targeting the SFRP5-SP1 axis in AF-related cardiac complications.

分泌型褐飞虱相关蛋白 5（SFRP5）是一种公认的心脏保护蛋白，但在心房颤动（房颤）中的表达量却减少了。本研究调查了 SFRP5 在心房颤动相关心脏纤维化和心肌细胞凋亡中的功能，探讨了其潜在的失调原因。利用 C57BL/6 小鼠、小鼠心脏成纤维细胞（CFs）和 HC-1 小鼠心房肌细胞系，通过血管紧张素Ⅱ（Ang Ⅱ）诱导房颤模型。临床患者血浆样本、模型小鼠血浆样本和 CF 培养上清液中 SFRP5 的水平持续下降。用重组 SFRP5 治疗可恢复其水平，减轻 Ang Ⅱ 诱导的小鼠房颤，改善心房组织纤维化和氧化应激。在体外，SFRP5 重组蛋白可抑制 CF 活化和纤维化相关标志物。研究发现 Sp1 转录因子（SP1）与 SFRP5 启动子结合，导致转录抑制。SP1敲除可恢复小鼠和CF中的SFRP5水平，从而抑制纤维化。此外，SP1敲除可减轻Ang Ⅱ诱导的HC-1细胞凋亡，但同时敲除SFRP5可抵消这种效应。总之，这项研究强调，SP1 在房颤期间通过转录抑制介导了 SFRP5 的丢失，导致纤维化和肌细胞凋亡。这些发现揭示了针对房颤相关心脏并发症的 SFRP5-SP1 轴的潜在治疗干预措施。

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引用次数: 0

Matrix Gla protein suppresses osteoblast senescence and promotes osteogenic differentiation by the PI3K-AKT signaling pathway 基质 Gla 蛋白通过 PI3K-AKT 信号通路抑制成骨细胞衰老并促进成骨细胞分化。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2024-11-12 DOI: 10.1016/j.yexcr.2024.114329

Min Zhang , Sha Liu , Yulin Chen , Yifa Chen , Jiaojiao He , Yuting Xia , Ya Yang

Age-related bone loss in mice is associated with senescent cell accumulation and reduced bone formation by osteoblasts. Matrix Gla protein (MGP), secreted by osteoblasts, is pivotal in regulating the bone extracellular matrix mineralization. Previous research has demonstrated that Mgp null mice exhibit osteopenia and fractures, and ultimately die prematurely. To elucidate the mechanisms underlying MGP's role of MGP in bone metabolism, we generated osteoblast-specific Mgp knockout (Mgp cKO) mice by crossing Mgp^fl/fl mice with Bglap-Cre mice. The study revealed that in 3-month-old Mgp cKO male mice, trabecular bone volume decreased, and the senescence marker protein p21 increased. Primary osteoblasts from Mgp cKO mice exhibited markers of DNA damage and senescence, such as increased γH2AX foci, p21, and senescence-associated β-galactosidase staining, as well as attenuated cellular proliferation and osteogenic differentiation abilities. In addition, bone marrow stromal cells' colony formation and spontaneous osteogenic ability were impaired in Mgp cKO mice, whereas osteoclastogenesis was enhanced. In vitro treatment with recombinant human MGP promotes osteogenesis in osteoblasts derived from Mgp cKO mice via the PI3K-AKT signaling pathway. Thus, our results suggest that MGP is protective by suppressing osteoblast senescence, offering new insights into potential therapeutic strategies for age-related osteoporosis.

小鼠与年龄相关的骨质流失与衰老细胞积累和成骨细胞骨形成减少有关。成骨细胞分泌的基质Gla蛋白（MGP）在调节骨细胞外基质矿化过程中起着关键作用。先前的研究表明，Mgp 缺失的小鼠会出现骨质增生和骨折，并最终过早死亡。为了阐明MGP在骨代谢中的作用机制，我们通过将Mgpfl/fl小鼠与Bglap-Cre小鼠杂交，产生了成骨细胞特异性Mgp基因敲除（Mgp cKO）小鼠。研究发现，3 个月大的 Mgp cKO 雄性小鼠骨小梁体积减少，衰老标志蛋白 p21 增加。Mgp cKO小鼠的原代成骨细胞表现出DNA损伤和衰老的标志物，如γH2AX病灶、p21和衰老相关的β-半乳糖苷酶染色增加，细胞增殖和成骨分化能力减弱。此外，Mgp cKO 小鼠骨髓基质细胞的集落形成和自发性成骨能力受损，而破骨细胞生成能力增强。体外处理重组人MGP可通过PI3K-AKT信号通路促进Mgp cKO小鼠成骨细胞的成骨。因此，我们的研究结果表明，MGP 通过抑制成骨细胞的衰老起到保护作用，为老年性骨质疏松症的潜在治疗策略提供了新的思路。

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引用次数: 0

METTL3/YTHDF1 stabilizes CORO6 expression promoting osteosarcoma progression through glycolysis METTL3/YTHDF1通过糖酵解稳定CORO6的表达，促进骨肉瘤的进展。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2024-11-12 DOI: 10.1016/j.yexcr.2024.114328

Xuzhou Liu , Wenchong Yu , Wei Song , Zhengqian Zhang , Benqiang Chen , Hongsheng Lin

This study investigates the role of CORO6 (Coronin 6) in the development of osteosarcoma. Osteosarcoma is a common malignant bone tumor in children and adolescents, characterized by rapid and irregular bone growth and a high risk of distant lung metastasis. CORO6 is a member of the Coronin family, known for its conserved WD40 repeat domain. This structure allows CORO6 to inhibit actin dynamics through interactions with F-actin and Arp2/3, thereby affecting the organization of the cytoskeleton. Our research found that in osteosarcoma patients, the levels of CORO6 are significantly elevated. Experimental observations showed that reducing the expression of CORO6 significantly inhibits the growth, migration, and invasion abilities of osteosarcoma cells. Moreover, in vivo experiments demonstrated that the absence of CORO6 effectively inhibits the growth of osteosarcoma in animal models. We also discovered that CORO6 promotes the proliferation, migration and invasion capabilities of osteosarcoma cells by activating the Wnt/β-catenin signaling pathway. Moreover, CORO6 plays a critical important role in glycolysis of osteosarcoma cells. Mechanically, we found that METTL3/YTHDF1 induced m6A modification of CORO6 mRNA promoted the expression of CORO6 by enhancing its stability. These findings offer new directions for the treatment of osteosarcoma, suggesting that CORO6 could be a novel prognostic biomarker and an effective therapeutic target for patients.

In summary, CORO6, as an oncogene, plays a key role in the development of osteosarcoma, providing a crucial theoretical basis for the development of new osteosarcoma treatment strategies.

这项研究探讨了CORO6（冠状病毒蛋白6）在骨肉瘤发病过程中的作用。骨肉瘤是儿童和青少年常见的恶性骨肿瘤，其特点是骨生长迅速且不规则，远处肺转移风险高。CORO6 是 Coronin 家族的成员，因其保守的 WD40 重复结构域而闻名。这种结构使CORO6能够通过与F-肌动蛋白和Arp2/3的相互作用抑制肌动蛋白的动力学，从而影响细胞骨架的组织。我们的研究发现，在骨肉瘤患者中，CORO6 的水平显著升高。实验观察表明，减少 CORO6 的表达可明显抑制骨肉瘤细胞的生长、迁移和侵袭能力。此外，体内实验表明，在动物模型中，CORO6 的缺失可有效抑制骨肉瘤的生长。我们还发现，CORO6 通过激活 Wnt/β-catenin 信号通路促进骨肉瘤细胞的增殖、迁移和侵袭能力。此外，CORO6 在骨肉瘤细胞的糖化过程中发挥着至关重要的作用。在机制上，我们发现 METTL3/YTHDF1 诱导的 CORO6 mRNA m6A 修饰通过增强 CORO6 的稳定性来促进其表达。这些发现为骨肉瘤的治疗提供了新的方向，提示CORO6可能是一种新的预后生物标志物和患者的有效治疗靶点。综上所述，CORO6作为一种癌基因，在骨肉瘤的发生发展中起着关键作用，为制定新的骨肉瘤治疗策略提供了重要的理论依据。

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引用次数: 0

Exosomes derived from diabetic microenvironment-preconditioned mesenchymal stem cells ameliorate nonalcoholic fatty liver disease and inhibit pyroptosis of hepatocytes 从糖尿病微环境预处理间充质干细胞中提取的外泌体可改善非酒精性脂肪肝并抑制肝细胞的嗜热性。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2024-11-07 DOI: 10.1016/j.yexcr.2024.114325

Anning Wang , Bing Li , Wanlu Su , HaiXia Zhang , Ruofan Hu , Yue Zhang , Jian Zhao , Rui Ren , Yiming Mu , Yu Cheng , Zhaohui Lyu

Aim

Pyroptosis, a type of programmed cell death, is a key mechanism underlying non-alcoholic fatty liver disease (NAFLD). Mesenchymal stem cell (MSC)-derived exosomes (MSC-Exos) have the potential to ameliorate NAFLD, an effect that is enhanced by curcumin preconditioning. We previously reported that diabetic microenvironment preconditioning enhances the secretion capacity and anti-inflammatory activity of MSCs. Therefore, we hypothesized that MSC-Exos would inhibit hepatocyte pyroptosis and thereby ameliorate NAFLD, and that diabetic microenvironment preconditioning would enhance these effects.

Methods

MSCs were preconditioned in a diabetic microenvironment (pMSCs). MSC-Exos and pMSC-Exos collected from MSCs or pMSCs were applied to methionine- and choline-deficient (MCD)-induced NAFLD mice and in vitro models involving induction with lipopolysaccharide or palmitic acid to mimic hepatic steatosis and injury. MCC950 treatment was used as a positive control. We analyzed the characteristics of NAFLD and pyroptosis markers. Protein profiles of MSC-Exos and pMSC-Exos were evaluated by label-free quantitative proteomics.

Results

In vivo, MSC-Exos partially attenuated inflammation and fibrosis, but not lipid deposition and NAFLD progression in the livers of NAFLD mice. pMSC-Exos significantly improved lipid metabolism, hepatic steatosis, inflammation, and fibrosis but also retarded the progression of NAFLD. Pyroptosis was upregulated in the liver of NAFLD mice. MSC-Exos and pMSC-Exos inhibited pyroptosis, and the effect of the latter was greater than that of the former. In vitro, MSC-Exos and pMSC-Exos ameliorated hepatocyte steatosis, lipid metabolism disorder, and inflammation, and pMSC-Exos exerted a greater inhibitory effect on hepatocyte pyroptosis than MSC-Exos did, which were remitted after inhibition of peroxiredoxin-1 (PRDX-1).

Conclusion

MSC-Exos ameliorated NAFLD and inhibited hepatocyte pyroptosis by downregulating the NLRP3/Caspase-1/GSDMD pathway, effects enhanced by pMSC-Exos, partly due to PRDX-1 upregulation.

目的：热核变性是一种程序性细胞死亡，是非酒精性脂肪肝（NAFLD）的一个关键机制。间充质干细胞（MSC）衍生的外泌体（MSC-Exos）具有改善非酒精性脂肪肝的潜力，姜黄素预处理可增强这种效果。我们曾报道，糖尿病微环境预处理可增强间充质干细胞的分泌能力和抗炎活性。因此，我们假设间充质干细胞-Exos能抑制肝细胞脓毒症，从而改善非酒精性脂肪肝，而糖尿病微环境预处理能增强这些效果：方法：间充质干细胞在糖尿病微环境（pMSCs）中进行预处理。方法：在糖尿病微环境（pMSCs）中对间充质干细胞进行预处理。将从间充质干细胞或pMSCs中收集的间充质干细胞-Exos和pMSC-Exos应用于蛋氨酸和胆碱缺乏（MCD）诱导的非酒精性脂肪肝小鼠和体外模型，这些模型涉及用脂多糖或棕榈酸诱导以模拟肝脏脂肪变性和损伤。MCC950 处理被用作阳性对照。我们分析了非酒精性脂肪肝的特征和脂肪变性标志物。通过无标记定量蛋白质组学评估了MSC-Exos和pMSC-Exos的蛋白质谱：在体内，间充质干细胞-Exos能部分减轻非酒精性脂肪肝小鼠肝脏的炎症和纤维化，但不能减轻脂质沉积和非酒精性脂肪肝的进展。非酒精性脂肪肝小鼠肝脏中的裂解酶上调。间充质干细胞-Exos和pMSC-Exos可抑制非酒精性脂肪肝小鼠肝脏的裂解，且后者的作用大于前者。在体外，间充质干细胞-Exos和pMSC-Exos能改善肝细胞脂肪变性、脂质代谢紊乱和炎症，pMSC-Exos对肝细胞脓毒症的抑制作用比间充质干细胞-Exos更大，在抑制过氧化物歧化酶-1（PRDX-1）后，肝细胞脓毒症得到缓解：结论：间充质干细胞-Exos可通过下调NLRP3/Caspase-1/GSDMD通路改善非酒精性脂肪肝并抑制肝细胞脓毒症。

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引用次数: 0

Downregulation of CTRP1 Reduces Radio-resistance in Glioblastoma Cells by Inhibiting the Expression of CD133 after X-ray and Carbon Ion Irradiation. 下调 CTRP1 可抑制 CD133 在 X 射线和碳离子照射后的表达，从而降低胶质母细胞瘤细胞的放射抗性

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2024-11-06 DOI: 10.1016/j.yexcr.2024.114292

Ke Huang, Luyao Wu, Dan Xv, Hong Zhang, Qiang Liu, Yi Xie

Glioblastomas (GBMs), the most prevalent primary malignant brain tumors, present significant challenges due to their invasive nature, high recurrence rates, and limited treatment options. Radiotherapy is a cornerstone in the management of GBMs; however, resistance to treatment poses a substantial obstacle. This study investigates the role of adipokine C1q/TNF-related protein 1 (CTRP1) in the radio-sensitivity of GBMs, utilizing both X-ray and carbon ion irradiation. Expression analyses revealed elevated CTRP1 and CD133 levels in GBMs tissues, which were associated with poor patient survival. Carbon ion irradiation demonstrated superior growth inhibition compared to X-ray, particularly in U87 (high CD133) cells. Moreover, CTRP1 expression increased following radiation exposure, especially after X-ray treatment. Knockdown of CTRP1 enhanced radio-sensitivity by reducing cell proliferation and increasing apoptosis, while exacerbating oxidative stress. Bioinformatics analysis revealed CTRP1's involvement in DNA damage repair pathways. Our findings establish a novel connection between CTRP1 and cellular radio-sensitivity. Targeting CTRP1, especially in U87 (high CD133) cells, enhances GBMs radio-sensitivity, offering potential therapeutic avenues.

胶质母细胞瘤（GBMs）是最常见的原发性恶性脑肿瘤，由于其侵袭性、高复发率和有限的治疗方案，给治疗带来了巨大挑战。放疗是治疗脑胶质瘤的基石，然而，耐药性是治疗的一大障碍。本研究利用X射线和碳离子照射，研究了脂肪因子C1q/TNF相关蛋白1（CTRP1）在GBMs放射敏感性中的作用。表达分析表明，GBMs 组织中的 CTRP1 和 CD133 水平升高与患者存活率低有关。与 X 射线相比，碳离子照射对生长的抑制作用更强，尤其是对 U87（高 CD133）细胞。此外，CTRP1的表达在辐照后增加，尤其是在X射线治疗后。通过减少细胞增殖和增加细胞凋亡，同时加剧氧化应激，敲除 CTRP1 增强了放射敏感性。生物信息学分析表明，CTRP1参与了DNA损伤修复途径。我们的研究结果在 CTRP1 与细胞放射敏感性之间建立了一种新的联系。靶向 CTRP1，尤其是 U87（高 CD133）细胞，可增强 GBM 的放射敏感性，从而提供潜在的治疗途径。

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引用次数: 0

AXL/GAS6 signaling governs differentiation of tumor-associated macrophages in breast cancer AXL/GAS6 信号调节乳腺癌中肿瘤相关巨噬细胞的分化。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2024-11-06 DOI: 10.1016/j.yexcr.2024.114324

Suman Purohit , Gunjan Mandal , Subir Biswas , Shauryabrota Dalui , Arnab Gupta , Sougata Roy Chowdhury , Arindam Bhattacharyya

Most epithelial cancers are infiltrated by prognostically relevant myelomonocytic cells. Immunosuppressive tumor associated macrophages (TAMs) and their precursor monocytic myeloid-derived suppressor cells (MDSCs) have previously been associated with worse outcomes in human breast cancer (BCa), yet the mechanism of immunosuppressive TAMs-polarization from myelomonocytic precursors is not completely understood. In this study, we show that persuaded AXL/GAS6 pathway alters macrophage phenotype from HLA-DR^highCD206^lowCD163^low classical phagocytic into HLA-DR^lowCD206^highCD163^high immunosuppressive ones with accelerated BCa progression, and increased angiogenesis signature and invasion ability of cancer cells at tumor beds. Notably, both AXL and GAS6 expressions are upregulated in human invasive breast carcinoma, with maximum expression in triple negative histology type. Mechanistically, we demonstrate that AXL/GAS6 signaling drives immunosuppression by governing increased immunosuppressive IL10 production while dampening IL-1β expression within the tumor microenvironment (TME) of BCa. Further, AXL/GAS6 signaling promotes angiogenesis through the activation of PI3K/AKT and NF-κB signaling pathways. Our results unveil role of AXL/GAS6 axis in the differentiation of TAMs, which governs malignant growth, and suggest that therapies that uncouple AXL/GAS6 axis may exhibit therapeutic opportunity for otherwise undruggable Triple Negative Breast Cancer (TNBC) patients.

大多数上皮癌都有预后相关的骨髓单核细胞浸润。免疫抑制性肿瘤相关巨噬细胞（TAMs）及其前体单核细胞-MDSCs曾与人类乳腺癌（BCa）的不良预后相关，但免疫抑制性TAMs从髓单核细胞前体极化的机制尚未完全明了。在这项研究中，我们发现AXL/GAS6通路会改变巨噬细胞的表型，使其从HLA-DRhighCD206-low CD163-low经典吞噬型转变为HLA-DR-lowCD206-highCD163-high免疫抑制型，从而加速BCa的进展，并增加肿瘤床的血管生成特征和癌细胞的侵袭能力。值得注意的是，AXL和GAS6在人类浸润性乳腺癌中均表达上调，在三阴性组织学类型中表达最高。从机理上讲，我们证明了 AXL/GAS6 信号通过调节免疫抑制 IL10 的产生，同时抑制 BCa 肿瘤微环境（TME）中 IL-1β 的表达，从而驱动免疫抑制。此外，AXL/GAS6 信号通过激活 PI3K/AKT 和 NF-κB 信号通路促进血管生成。我们的研究结果揭示了AXL/GAS6轴在TAMs分化过程中的作用，TAMs分化决定着恶性肿瘤的生长，这表明解除AXL/GAS6轴的疗法可能会给原本无法治愈的三阴性乳腺癌（TNBC）患者带来治疗机会。

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引用次数: 0

SIRT3 mitigates high glucose-induced damage in retinal microvascular endothelial cells via OPA1-mediated mitochondrial dynamics. SIRT3 通过 OPA1 介导的线粒体动力学减轻高血糖诱导的视网膜微血管内皮细胞损伤。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2024-11-02 DOI: 10.1016/j.yexcr.2024.114320

Jiemei Shi, Min Liu, Haohao Zhu, Chunhui Jiang

Oxidative stress in endothelial cells is pivotal in diabetic retinopathy (DR), with mitochondrial homeostasis being crucial to mitigate this stress. This study explored the roles of mitochondrial sirtuins (SIRTs) in high glucose (HG)-induced oxidative stress, related endothelial impairment, and mitochondrial homeostasis damage in rat retinal microvascular endothelial cells (RMECs). RMECs were cultured under HG or equivalent osmotic conditions. Cell viability was assessed using the Cell Counting Kit-8 assay, whereas cell death and survival were determined via calcein-AM/propidium iodide double staining. Reactive oxygen species (ROS) levels were measured using 2',7'-dichlorofluorescein fluorescence. Expression of mitochondrial SIRTs3-5 and key mitochondrial homeostasis molecules was quantified by the quantitative real-time polymerase chain reaction and confirmed by western blotting. Mitochondrial morphology was evaluated using electron microscopy and the MitoTracker fluorescent probe. A SIRT3-overexpressing RMEC line was constructed to assess the role of SIRT3 in oxidative stress and mitochondrial dynamics. After 48 h of HG exposure, cell viability was significantly reduced, with a concomitant increase in cell death and ROS levels, alongside a marked decrease in SIRT3 expression and molecules associated with mitochondrial dynamics. SIRT3 overexpression reversed these effects, particularly increasing the mitochondrial fusion-related molecule, optic atrophy 1 (OPA1). However, the OPA1 inhibitor, MYLS22, blocked the protective effect of SIRT3, leading to more dead cells, a higher ROS level, and intensified mitochondrial fragmentation. These results suggest that SIRT3 is involved in HG-induced imbalanced mitochondrial dynamics of endothelial cells in DR, potentially through the OPA1 pathway.

内皮细胞的氧化应激在糖尿病视网膜病变（DR）中起着关键作用，而线粒体的平衡对缓解这种应激至关重要。本研究探讨了线粒体sirtuins（SIRTs）在高糖（HG）诱导的氧化应激、相关内皮损伤和大鼠视网膜微血管内皮细胞（RMECs）线粒体稳态损伤中的作用。在 HG 或同等渗透条件下培养 RMECs。使用细胞计数试剂盒-8 评估细胞活力，而细胞死亡和存活则通过钙黄绿素-AM/碘化丙啶双重染色来确定。活性氧（ROS）水平用 2',7'-二氯荧光素荧光测定。线粒体 SIRTs3-5 和关键线粒体稳态分子的表达通过定量实时聚合酶链反应进行量化，并通过 Western 印迹法进行确认。使用电子显微镜和 MitoTracker 荧光探针对线粒体形态进行了评估。为了评估 SIRT3 在氧化应激和线粒体动力学中的作用，构建了一个 SIRT3 表达过高的 RMEC 株系。暴露于 HG 48 小时后，细胞活力明显降低，同时细胞死亡和 ROS 水平增加，SIRT3 表达和线粒体动力学相关分子明显减少。SIRT3 的过量表达可逆转这些影响，尤其是线粒体融合相关分子视神经萎缩 1（OPA1）的增加。然而，OPA1 抑制剂 MYLS22 阻断了 SIRT3 的保护作用，导致更多的死亡细胞、更高的 ROS 水平和线粒体破碎加剧。这些结果表明，SIRT3 可能通过 OPA1 途径参与了 HG 诱导的 DR 内皮细胞线粒体动力学失衡。

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引用次数: 0

Exosomal lncRNA Mir100hg derived from cancer stem cells enhance glycolysis and promote metastasis of melanoma through miR-16-5p and miR-23a-3p 来自癌症干细胞的外泌体 lncRNA Mir100hg 通过 miR-16-5p 和 miR-23a-3p 增强糖酵解并促进黑色素瘤的转移。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2024-11-01 DOI: 10.1016/j.yexcr.2024.114319

Jiyu Tan , Yao Tang , Bowen Li , Lei Shi , Yuhan Zhang , Yuting Chen , Yan Chen , Jie Li , Meng Xiang , Yufeng Zhou , H. Rosie Xing , Jianyu Wang

Increasing evidence demonstrate that the significant role of long non-coding RNA (lncRNA) in metastasis and the remodeling of the tumor microenvironment. However, the precise mechanisms of lncRNAs in cancer metastasis are still poorly understood. The function of lncRNA-Mir100hg in melanoma and its involvement in mediating communication between tumor stem cells and non-stemness tumor cells remains unknown. We found that Mir100hg is upregulated in melanoma stem cells (CSCs) known as OLSD. Furthermore, Mir100hg can be transferred from OLSD to non-stem cancer cells (OL) through exosomes. Once Mir100hg enters OL cells, it operates through a competitive endogenous RNA (ceRNA) mechanism. It competes with microRNAs (miR-16-5p and miR-23a-3p) by binding to them, thus preventing these miRNAs from targeting their mRNAs. As a result, the expression of glycolysis-related mRNA was restored. This ultimately enhances the metastatic capability of OL cells. In summary, our study uncovers a network used by CSCs to transfer their high metastatic activity to non-stem cancer cells through the exosomal Mir100hg. This mechanism sheds new light on the communication between heterogeneous cancer cell populations in melanoma. Importantly, it provides novel insights into the role of lncRNAs in cancer metastasis and highlights the significance of the tumor microenvironment in facilitating metastasis.

越来越多的证据表明，长非编码 RNA（lncRNA）在癌症转移和肿瘤微环境重塑中发挥着重要作用。然而，人们对 lncRNA 在癌症转移中的确切机制仍知之甚少。lncRNA-Mir100hg在黑色素瘤中的功能及其参与介导肿瘤干细胞和非干性肿瘤细胞之间的交流仍是未知数。我们发现，Mir100hg在被称为OLSD的黑色素瘤干细胞（CSCs）中上调。此外，Mir100hg可通过外泌体从OLSD转移到非干细胞癌细胞（OL）。Mir100hg 进入 OL 细胞后，会通过竞争性内源性 RNA（ceRNA）机制发挥作用。它通过与 microRNA（miR-16-5p 和 miR-23a-3p）结合来与它们竞争，从而阻止这些 miRNA 靶向它们的 mRNA。因此，糖酵解相关 mRNA 的表达得到了恢复。这最终增强了 OL 细胞的转移能力。总之，我们的研究揭示了干细胞通过外泌体 Mir100hg 将其高转移活性转移到非干癌细胞的网络。这一机制为黑色素瘤中异质癌细胞群之间的交流提供了新的线索。重要的是，它为lncRNA在癌症转移中的作用提供了新的见解，并强调了肿瘤微环境在促进转移中的重要作用。

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引用次数: 0

Targeting of ubiquitination and degradation of KLF15 by E3 ubiquitin ligase KBTBD7 regulates LPS-induced septic brain injury in microglia E3 泛素连接酶 KBTBD7 对 KLF15 泛素化和降解的靶向调节 LPS 诱导的小胶质细胞脓毒性脑损伤。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2024-11-01 DOI: 10.1016/j.yexcr.2024.114317

Wei Shen, Xuzhong Zhang, Min Tang, Wei Chen, Ying Wang, Haoquan Zhou

Septic brain injury is a serious disease of the central nervous system that involves inflammation. Kelch repeat and BTB domain containing 7 (KBTBD7), an E3 ubiquitin ligase, is demonstrated to facilitate the pathological changes of various diseases, but its impact on septic brain injury is unclear. In this study, a rat model of septic brain injury was induced by cecal ligation and puncture (CLP). The neurobehavioral score and survival rate of CLP group were worse than those of sham group. In addition, CLP was found to evoke microglia activation, increase inflammation, induce the activation of NLRP3 inflammasome and NF-κB signaling pathway, and upregulate KBTBD7 expression. Immunofluorescence revealed strong positive KBTBD7 staining in CLP rat microglia. Furthermore, primary microglia were exposed to lipopolysaccharide (LPS) to explore the role and mechanism of KBTBD7. The results showed that KBTBD7 expression was increased in LPS-treated microglia. Knockdown of KBTBD7 markedly inhibited LPS-induced proinflammatory cytokine release, as well as the activation of NLRP3 inflammasome and NF-κB signaling pathway. The downstream molecular mechanism of KBTBD7 was then mined. Notably, co-immunoprecipitation (co-IP) results confirmed that KBTBD7 was a novel interacting protein of KLF transcription factor 15 (KLF15) and acted as an E3 ubiquitin ligase that catalyzed the ubiquitination degradation of KLF15 through the ubiquitin-proteasome system. Moreover, recovery experiment data suggested that KLF15 knockdown abolished the anti-inflammatory role of KBTBD7 knockdown in microglia, implying that KLF15 influenced the function of KBTBD7. Taken together, our results reveal a novel KBTBD7-KLF15 signal transduction pathway involved in septic brain injury and provide a potential therapeutic strategy for its treatment.

脓毒性脑损伤是一种涉及炎症的严重中枢神经系统疾病。Kelch repeat and BTB domain containing 7 (KBTBD7) 是一种 E3 泛素连接酶，已被证实能促进多种疾病的病理变化，但它对败血症性脑损伤的影响尚不清楚。本研究通过盲肠结扎和穿刺（CLP）诱导大鼠脓毒性脑损伤模型。CLP组的神经行为评分和存活率均低于假组。此外，CLP还可诱发小胶质细胞活化，增加炎症反应，诱导NLRP3炎性体和NF-κB信号通路的活化，并上调KBTBD7的表达。免疫荧光显示，CLP 大鼠小胶质细胞中的 KBTBD7 染色呈强阳性。此外，还将原代小胶质细胞暴露于脂多糖（LPS），以探讨 KBTBD7 的作用和机制。结果显示，KBTBD7 在 LPS 处理的小胶质细胞中表达增加。敲除 KBTBD7 能显著抑制 LPS 诱导的促炎细胞因子释放，以及 NLRP3 炎性体和 NF-κB 信号通路的激活。随后，研究人员对 KBTBD7 的下游分子机制进行了挖掘。值得注意的是，共免疫沉淀（co-IP）结果证实，KBTBD7是KLF转录因子15（KLF15）的新型互作蛋白，并作为E3泛素连接酶，通过泛素-蛋白酶体系统催化KLF15的泛素化降解。此外，恢复实验数据表明，KLF15敲除可取消KBTBD7敲除在小胶质细胞中的抗炎作用，这意味着KLF15影响了KBTBD7的功能。综上所述，我们的研究结果揭示了一种参与脓毒性脑损伤的新型 KBTBD7-KLF15 信号转导通路，并为脓毒性脑损伤的治疗提供了一种潜在的治疗策略。

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Effect of isoproterenol, a β-adrenergic agonist, on the differentiation of insulin-producing pancreatic β cells derived from human pluripotent stem cells 异丙肾上腺素（一种β-肾上腺素能激动剂）对人多能干细胞衍生的胰岛素分泌β细胞分化的影响。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2024-11-01 DOI: 10.1016/j.yexcr.2024.114307

Hye Ryeong Jun , Yang Hee Kim , Ji Eun Moon , Sehui Jeong , Han Se Goh , Minh Hien Hoang , Yu Na Lee , Hyemin Jeong , In kyong Shim , Song Cheol Kim

Research on islet replacement through the differentiation of functionally matured insulin-producing β-like cells for the treatment of diabetes presents a significant challenge. Neural signals in β cell differentiation significantly impact the pancreatic microenvironment in glucose metabolism, but they are not fully understood. In this study, isoproterenol, a β adrenoreceptor agonist, was introduced into pancreatic progenitor cells, derived from human pluripotent stem cells in vitro, undergoing endocrine differentiation, using 2-dimensional (2D) and 3-dimensional (3D) differentiation protocols. This resulted in increased insulin and C-peptide secretion, along with elevated expression of key pancreatic beta cell transcription factors, including PDX-1, NKX6.1, and MAFA, and improved function, demonstrated by increased responsiveness to glucose determined via a glucose-stimulated insulin secretion test. Moreover, RNA transcriptome analysis of isoproterenol-treated endocrine progenitors facilitated the identification of biological pathways and genes that contribute to mature beta cell differentiation efficiency correlated with neural signals, such as adrenoceptor beta 1, calcium/calmodulin dependent protein kinase II alpha, phospholipase C delta 4, and neurotrophic receptor tyrosine kinase 1. Among those genes, calcium/calmodulin dependent protein kinase II alpha was suggested as the most notable gene involved in the isoproterenol mechanism through inhibitor assays. This study illustrates that isoproterenol significantly enhances endocrine differentiation and underscores its effects on stem cell-derived beta cell maturation, emphasizing its therapeutic potential for the treatment of diabetes.

通过分化功能成熟的胰岛素分泌β样细胞来替代胰岛治疗糖尿病的研究是一项重大挑战。β细胞分化过程中的神经信号对葡萄糖代谢过程中的胰腺微环境有重大影响，但人们对这些信号还不完全了解。在这项研究中，使用二维（2D）和三维（3D）分化方案，将异丙肾上腺素（一种β肾上腺素受体激动剂）导入体外进行内分泌分化的胰腺祖细胞（来源于人类多能干细胞）。这导致了胰岛素和C肽分泌的增加，以及关键胰腺β细胞转录因子（包括PDX-1、NKX6.1和MAFA）表达的升高，并通过葡萄糖刺激胰岛素分泌测试确定对葡萄糖反应性的增加，从而改善了功能。此外，对异丙托肾上腺素处理的内分泌祖细胞进行的 RNA 转录组分析有助于确定有助于成熟β细胞分化效率的生物通路和与神经信号相关的基因，如肾上腺素受体β1、钙/钙调蛋白依赖性蛋白激酶 II alpha、磷脂酶 C delta 4 和神经营养受体酪氨酸激酶 1。在这些基因中，钙/钙调蛋白依赖性蛋白激酶 II alpha 被认为是通过抑制剂实验参与异丙托品醇机制的最显著基因。这项研究表明，异丙托肾上腺素能显著促进内分泌分化，并强调了它对干细胞衍生的β细胞成熟的影响，强调了它治疗糖尿病的潜力。

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