Experimental cell research最新文献_第5页

Glycolysis inhibition and AMPK activation: a critical role of CTRP1 deficiency in the treatment of hypoxia-induced pulmonary hypertension 糖酵解抑制和AMPK激活：CTRP1缺乏在治疗缺氧性肺动脉高压中的关键作用

IF 3.5 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-11-22 DOI: 10.1016/j.yexcr.2025.114838

Yu Zhang , Weiqiang Chen , Kai Yang, Tao Hao, Wenfeng Cao, Shuqiang Dong

In pulmonary hypertension (PH), metabolic enhancement of glycolysis drives a hyperproliferative and apoptosis-resistant phenotype in pulmonary artery smooth muscle cells (PASMCs), which is a key pathological process leading to pulmonary vascular remodeling. We aimed to uncover the role of CTRP1, a gene responsible for regulating glycolysis, in the modulation of PH-related pathogenesis. In the PH mouse and cell models established by hypoxia stimulation, the regulatory mechanism of CTRP1 deletion on the pathological characteristics of PH was explored. CTRP1 levels were significantly upregulated in PH mice, accompanied by an abnormal increase in lactate production and glycolysis-related key protein expressions (HK2 and PDHK1). Inhibition of CTRP1 markedly improved pulmonary artery pressure and right ventricular function in PH mice by reducing glycolysis levels. In vitro experiments further observed that CTRP1 knockdown suppressed the hypoxia-induced hyperproliferation and anti-apoptosis phenotype of PASMCs, with inhibition of glycolysis. Mechanically, downregulated CTRP1 resulted in p-AMPK activation and p-AKT/mTOR inhibition. This beneficial effect was reversed by AMPKα2 deficiency. Overall, CTRP1 deficiency reverses the hypoxia-induced hyperproliferation and antiapoptotic capacity of PASMCs by weakening glycolysis. These results provide evidence for CTRP1 as a potential therapeutic target in PH.

在肺动脉高压（PH）中，糖酵解的代谢增强驱动肺动脉平滑肌细胞（PASMCs）的超增殖和抗凋亡表型，这是导致肺血管重构的关键病理过程。我们旨在揭示CTRP1（一个负责调节糖酵解的基因）在ph相关发病机制中的作用。在缺氧刺激建立的PH小鼠和细胞模型中，探讨CTRP1缺失对PH病理特征的调控机制。PH小鼠CTRP1水平显著上调，乳酸生成和糖酵解相关关键蛋白（HK2和PDHK1）表达异常增加。抑制CTRP1通过降低糖酵解水平显著改善PH小鼠肺动脉压和右心室功能。体外实验进一步发现，CTRP1敲低可抑制缺氧诱导的PASMCs过度增殖和抗凋亡表型，并抑制糖酵解。机制上，CTRP1下调导致p-AMPK活化和p-AKT/mTOR抑制。AMPKα2缺乏逆转了这种有益作用。总的来说，CTRP1缺乏通过削弱糖酵解来逆转缺氧诱导的PASMCs的过度增殖和抗凋亡能力。这些结果为CTRP1作为PH的潜在治疗靶点提供了证据。

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引用次数: 0

Fbxo45 promotes cell viability, invasion and sunitinib resistance of clear cell renal cell carcinoma by targeting Erbin Fbxo45通过靶向Erbin促进透明细胞肾细胞癌的细胞活力、侵袭和舒尼替尼耐药性。

IF 3.5 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-11-22 DOI: 10.1016/j.yexcr.2025.114839

Xueshan Pan , Ke Yu , Kai Chen , Jiao Wang , Zheng Huang , Jiewen Wang , Tong Cao , Jia Ma

The mechanisms underlying the development and progression of clear cell renal cell carcinoma (ccRCC) and its sunitinib resistance are elusive. Fbxo45 is a member of the F-box protein family that has been demonstrated to participate in tumorigenesis. However, the role of Fbxo45 in ccRCC progression has not been characterized. This study aims to investigate the biological functions and molecular mechanism of Fbxo45 in ccRCC progression. We found that Fbxo45 knockdown inhibited the viability and motility of ccRCC cells, while Fbxo45 overexpression resulted in the opposite phenotype. Ectopic expression of Fbxo45 promoted tumor growth in mice. Fbxo45 expression was negatively correlated with Erbin expression, which has been reported to mediate anti-tumor activities in ccRCC. Furthermore, Fbxo45 facilitated ccRCC cell viability and motility by inhibiting Erbin. Notably, Fbxo45 upregulation reduced sunitinib sensitivity in ccRCC cells. Our results suggest that Fbxo45 could be a potential target for ccRCC treatment and sunitinib resistance.

透明细胞肾细胞癌（ccRCC）的发生和进展及其舒尼替尼耐药性的机制尚不清楚。Fbxo45是F-box蛋白家族的一员，已被证明参与肿瘤发生。然而，Fbxo45在ccRCC进展中的作用尚未明确。本研究旨在探讨Fbxo45在ccRCC进展中的生物学功能和分子机制。我们发现Fbxo45敲低抑制了ccRCC细胞的活力和运动性，而Fbxo45过表达导致相反的表型。Fbxo45异位表达促进小鼠肿瘤生长。Fbxo45的表达与Erbin的表达呈负相关，有报道称Erbin介导ccRCC的抗肿瘤活性。Fbxo45通过抑制Erbin促进ccRCC细胞活力和运动。值得注意的是，Fbxo45的上调降低了ccRCC细胞对舒尼替尼的敏感性。我们的结果表明Fbxo45可能是ccRCC治疗和舒尼替尼耐药的潜在靶点。

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引用次数: 0

Elucidating the impact of trans-ned-19 on two-Pore channel 2 mutants of Dictyostelium: changes in intracellular calcium levels and subsequent effect on autophagic flux 阐明反式ned -19对双孔通道2突变体Dictyostelium的影响：细胞内钙水平的变化及其对自噬通量的影响

IF 3.5 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-11-20 DOI: 10.1016/j.yexcr.2025.114829

Ashima Thakur, Shweta Saran

Two-Pore Channel 2 (TPC2) are calcium (Ca²⁺) release channels regulated by Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP), primarily localized to the endolysosomal system. TPC2 regulates diverse Ca²⁺-dependent processes such as cell proliferation, development, migration, and autophagy. In the protist Dictyostelium discoideum, intracellular Ca²⁺ levels and autophagy are key determinants of cell-fate, particularly for stalk cell differentiation. To investigate the involvement of DdTPC2 in NAADP-mediated Ca²⁺ signalling, we treated wild-type (Ax2) and various tpc2 mutant strains, like overexpressors (tpc2^OE), knockout (tpc2^-), and rescue (tpc2^Res), with both NAADP-AM (NAADP agonist) and trans-Ned-19 (NAADP antagonist). Our findings show that trans-Ned-19 does not specifically inhibit DdTPC2 activity during the proliferative (vegetative) stage, but acts as a specific TPC2 inhibitor in freshly starved cells or during multicellular development, as the phenotypes displayed by trans-Ned-19 treated Ax2 cells were similar to tpc2^- cells. Trans-Ned-19 colocalizes with lysosomes, consistent with the subcellular distribution of TPC2. Fluid-phase endocytosis was reduced in tpc2^- cells, implicating TPC2 in endolysosomal function. Intracellular calcium measurements demonstrated that NAADP regulates Ca²⁺ signalling through TPC2, as both NAADP-AM and trans-Ned-19 significantly modulated intracellular free Ca²⁺ levels in TPC2-expressing strains. Notably, trans-Ned-19 treatment reduced autophagic flux in Ax2 cells to tpc2^- levels, demonstrating a positive correlation between TPC2 and autophagic flux, further supported by rapamycin and 3-methyladenine treatment results. This study highlights TPC2 as a pivotal regulator of NAADP-mediated Ca²⁺ signalling and autophagy in Dictyostelium, with broad implications for understanding these processes in higher eukaryotes.

双孔通道2 （TPC2）是由烟酸腺嘌呤二核苷酸磷酸（NAADP）调节的钙（Ca2+）释放通道，主要定位于内溶酶体系统。TPC2调节多种Ca2+依赖性过程，如细胞增殖、发育、迁移和自噬。在原生体盘状体中，细胞内Ca2+水平和自噬是细胞命运的关键决定因素，特别是在茎细胞分化中。为了研究DdTPC2在NAADP介导的Ca2+信号传导中的作用，我们用NAADP- am （NAADP激动剂）和反式内酯-19 （NAADP拮抗剂）处理野生型（Ax2）和各种tpc2突变株，如过表达型（tpc2OE）、敲除型（tpc2-）和挽救型（tpc2Res）。我们的研究结果表明，在增殖（营养）阶段，反式ned -19并不特异性抑制DdTPC2的活性，但在新鲜饥饿细胞或多细胞发育期间，反式ned -19作为一种特异性的TPC2抑制剂，因为反式ned -19处理的Ax2细胞表现出与TPC2 -细胞相似的表型。trans-Ned-19与溶酶体共定位，与TPC2的亚细胞分布一致。tpc2-细胞的液相内吞作用减少，暗示tpc2参与内溶酶体功能。细胞内钙测量表明，NAADP通过TPC2调节Ca2+信号，因为NAADP- am和反式ned -19显著调节表达TPC2的菌株的细胞内游离Ca2+水平。值得注意的是，trans-Ned-19处理使Ax2细胞的自噬通量降低到tpc2-水平，表明tpc2与自噬通量呈正相关，雷帕霉素和3-甲基腺嘌呤处理结果进一步支持了这一结果。这项研究强调了TPC2作为naadp介导的Ca2+信号传导和Dictyostelium自噬的关键调节剂，对理解高等真核生物的这些过程具有广泛的意义。

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引用次数: 0

Metformin inhibits heterotopic ossification of mouse Achilles tendon by inhibiting the Nr4a1/Wnt/β-catenin signaling pathway 二甲双胍通过抑制Nr4a1/Wnt/β-catenin信号通路抑制小鼠跟腱异位骨化。

IF 3.5 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-11-14 DOI: 10.1016/j.yexcr.2025.114824

Danxia Zheng , Mengyu Jiang , Wenjie Wang , Jianglin Yu , Xiaoyang Qi , Yixin Chen

Heterotopic ossification is a pathological process characterized by aberrant bone formation in soft tissues, leading to joint pain, stiffness, functional impairment, and poor quality of life. We used a mouse Achilles tendon heterotopic ossification model and in vitro tendon-derived stem cells (TDSCs) assays to identify the molecular mechanism of metformin in prevention of heterotopic ossification. Metformin significantly attenuated heterotopic ossification, reducing ectopic bone volume and osteogenic gene expression. In vitro, metformin inhibited TDSCs osteogenic differentiation in a dose-dependent manner, decreasing calcium nodule deposition and osteogenic marker expression. Transcriptomic analysis revealed downregulated Nr4a1 expression in metformin-treated heterotopic ossification samples; in vitro experiments confirmed that Nr4a1 activation enhances TDSCs osteogenesis, and Nr4a1 knockdown suppresses osteogenesis. Metformin also reduced Wnt4 and β-catenin expression, suggesting that Nr4a1 promotes heterotopic ossification by positively regulating Wnt/β-catenin signaling. In sum, metformin downregulates expression of Nr4a1 in TDSCs, which suppresses osteogenic differentiation by inhibiting Wnt/β-catenin signaling. As a mediator of TDSC osteogenic differentiation, Nr4a1 may be a therapeutic target for heterotopic ossification.

异位骨化是一种病理过程，其特征是软组织骨形成异常，导致关节疼痛、僵硬、功能障碍和生活质量差。我们采用小鼠跟腱异位骨化模型和体外肌腱源性干细胞（tdsc）实验来确定二甲双胍预防异位骨化的分子机制。二甲双胍显著减弱异位骨化，减少异位骨体积和成骨基因表达。在体外，二甲双胍以剂量依赖的方式抑制tdsc的成骨分化，减少钙结节沉积和成骨标志物的表达。转录组学分析显示，二甲双胍处理的异位骨化样本中Nr4a1表达下调；体外实验证实Nr4a1激活促进tdsc成骨，而Nr4a1敲低抑制成骨。二甲双胍还降低了Wnt4和β-catenin的表达，表明Nr4a1通过正向调节Wnt/β-catenin信号通路促进异位骨化。综上所述，二甲双胍下调tdsc中Nr4a1的表达，通过抑制Wnt/β-catenin信号传导抑制成骨分化。作为TDSC成骨分化的中介，Nr4a1可能是异位骨化的治疗靶点。

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引用次数: 0

lincRNA-p21 drives apoptosis and calcification of vascular smooth muscle cell via small extracellular vesicles under hyperphosphatemic conditions in chronic kidney disease LincRNA-p21在慢性肾病高磷血症条件下通过细胞外小泡驱动血管平滑肌细胞凋亡和钙化。

IF 3.5 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-11-13 DOI: 10.1016/j.yexcr.2025.114779

Jianbing Hao, Siyu Wang, Lirong Hao

Vascular calcification, a major contributor to cardiovascular morbidity, involves pathological osteogenic transdifferentiation of vascular smooth muscle cell (VSMC) under hyperphosphatemic conditions in chronic kidney disease. This study investigates the role of lincRNA-p21 and small extracellular vesicles in phosphate-induced mouse aortic smooth muscle cell (MASMC) calcification. Exposure to phosphate (2.6 mmol/L) triggered time-dependent calcification, characterized by enhanced calcium deposition, endoplasmic reticulum remodeling, and intracellular calcium accumulation (p < 0.05 vs. 24/48-h controls). Concurrently, the expression of osteogenic markers (BGP, OCN, and OPN) and lincRNA-p21 was significantly upregulated, whereas the expression of contractile phenotype-specific markers (SM22α, SM-MHC, and SM α-actin) was markedly down-regulated. This pattern of gene expression was correlated with MASMC osteogenic transdifferentiation. Small extracellular vesicles isolated from phosphate-treated MASMC exhibited elevated lincRNA-p21 levels (p < 0.05) and induced calcification and apoptosis in recipient cells, suggesting small extracellular vesicles-mediated propagation of calcific signals. Functional studies demonstrated that lincRNA-p21 overexpression exacerbated calcification, apoptosis, and osteogenic marker expression, while its knockdown attenuated these effects (p < 0.05). Time-course analyses revealed lincRNA-p21 dynamically regulates small extracellular vesicles secretion, calcium accumulation, and apoptotic pathways, acting as a molecular switch driving phosphate-induced calcification. These findings establish lincRNA-p21 as a critical mediator of MASMC calcification via small extracellular vesicles-dependent mechanisms, offering insights into therapeutic strategies for vascular calcification.

血管钙化是心血管疾病的主要致病因素，涉及慢性肾病患者在高磷血症条件下血管平滑肌细胞（VSMC）的病理性成骨转分化。本研究探讨了lincRNA-p21和细胞外小泡在磷酸盐诱导的小鼠主动脉平滑肌细胞（MASMC）钙化中的作用。暴露于磷酸盐（2.6 mmol/L）触发了时间依赖性钙化，其特征是钙沉积增强、内质网重塑和细胞内钙积累(p

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引用次数: 0

ANXA1 inhibits ferroptosis in HUVECs via negative regulation of ferritinophagy ANXA1通过负调控铁蛋白自噬抑制HUVECs铁凋亡。

IF 3.5 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-11-13 DOI: 10.1016/j.yexcr.2025.114825

Jingjing Wang , Jinqiu Jia , Zitong Cao , Zeming Cai , Kai Zhang , Jin He , Chunyan Wu , Zuo Wang

Ferroptosis is a phospholipid peroxidation-mediated and iron-dependent cell death form. Inhibiting ferroptosis is a promising strategy for the prevention and treatment of cardiovascular diseases. Annexin A1 (ANXA1) is an endogenous anti-inflammatory mediator that plays an important regulatory role in cardiovascular diseases such as atherosclerosis, especially in inflammation suppression, protecting the heart from injury, and regulating vascular function. Ac2-26 is a synthetic peptide derived from the N-terminal 26 amino acids of ANXA1, which retains its anti-inflammatory properties. However, the regulatory mechanism of ANXA1 in atherosclerosis (AS) is not yet fully understood. This study aims to explore the specific role of ferroptosis in HUVECs and demonstrate that ANXA1 can disrupt ferritinophagy and protect endothelial function. Treatment with Ac2-26 or ANXA1-overexpressing HUVECs alleviated RSL3-induced endothelial cell dysfunction and inhibited lipid peroxidation, as evidenced by a reduction in ferrous ion levels and upregulation of GPX4, FTH1, and SLC7A11 protein expression, along with a downregulation of LC3-II and NCOA4 expression. In contrast, knockdown of ANXA1 in HUVECs failed to suppress the outcome of ferroptosis. Furthermore, co-immunoprecipitation analysis revealed that ANXA1 overexpression prevented ferritin degradation by disrupting the NCOA4-FTH1 protein-protein interaction, reducing the bioavailability of intracellular ferrous ions, thereby blocking ferroptosis. In conclusion, our findings identify a novel mechanism, showing that ANXA1 can inhibit ferroptosis via ferritinophagy, thereby alleviating endothelial dysfunction, which may provide a new therapeutic avenue for AS.

铁死亡是一种磷脂过氧化介导的铁依赖性细胞死亡形式。抑制铁下垂是预防和治疗心血管疾病的一种很有前途的策略。Annexin A1 （ANXA1）是一种内源性抗炎介质，在动脉粥样硬化等心血管疾病中发挥重要的调节作用，特别是在炎症抑制、保护心脏免受损伤、调节血管功能等方面。Ac2-26是由ANXA1的n端26个氨基酸合成的肽，保留了抗炎特性。然而，ANXA1在动脉粥样硬化（AS）中的调节机制尚不完全清楚。本研究旨在探讨铁凋亡在huvec中的具体作用，并证明ANXA1可以破坏铁蛋白吞噬，保护内皮功能。Ac2-26和anxa1过表达的HUVECs可以缓解rsl3诱导的内皮细胞功能障碍，抑制脂质过氧化，结果表明，亚铁离子水平降低，GPX4、FTH1和SLC7A11蛋白表达上调，LC3和NCOA4表达下调。相反，在huvec中敲低ANXA未能抑制铁下垂的结果。此外，共免疫沉淀分析显示，ANXA1过表达通过破坏NCOA4-FTH1蛋白-蛋白相互作用，降低细胞内铁离子的生物利用度，从而阻止铁蛋白的降解，从而阻止铁凋亡。总之，我们的研究发现了一种新的机制，表明ANXA1可以通过铁蛋白噬抑制铁下垂，从而减轻内皮功能障碍，这可能为AS的治疗提供新的途径。

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引用次数: 0

Augmenter of liver regeneration alleviates lipid accumulation in MASLD through ATF3-CD36 axis 肝再生增强剂通过ATF3-CD36轴缓解MASLD的脂质积累。

IF 3.5 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-11-13 DOI: 10.1016/j.yexcr.2025.114823

Weichun Xiao , Yuan Wu , Ling Qin , Jing Zhang , Wei An

Metabolic dysfunction-associated steatotic liver disease (MASLD) is characterized by excessive lipid deposition in hepatocytes. Augmenter of liver regeneration (ALR, encoded by the Gfer gene), a survival factor in liver cells, protects the liver against various injuries, whereas downregulation of ALR expression accelerates MASLD progression in mice. CD36 functions as a long chain free fatty acid transporter, playing a crucial role during pathogenesis of MASLD. Herein, the expression level of CD36 was found to significantly decrease in ALR-transfected (ALR-Tx) HepG2 cells and upregulated in shRNA-ALR (shALR) cells after oleic acid/palmitic acid treatment. Furthermore, CD36 expression was markedly elevated in MASLD model mice with heterozygous Gfer knockdown (Gfer^+/−) fed a high-fat diet,as well as model mice with liver-specific Gfer knockout (Gfer-CKO). Lipid uptake increased and lipophagy was suppressed in shALR HepG2 cells, and conversly, lipid uptake reduced and lipophagy was promoted in ALR-Tx HepG2 cells. After downregulation of CD36 expression by siRNA-CD36 in shALR cells, lipid accumulation markedly decreased, and lipophagy was stimulated. Further, the mRNA and protein expression of ATF3 (Activating Transcription Factor 3) were found to substantially elevated and decreased in ALR-Tx cells and shALR cells respectively, and CD36 expression was induced after ATF3 was knock down. Our results thus suggest that lack of ALR aggravates lipid deposition in MASLD partly through the ATF3-CD36 axis.

代谢功能障碍相关脂肪变性肝病（MASLD）的特征是肝细胞中脂质沉积过多。肝再生增强因子（ALR，由Gfer基因编码）是肝细胞中的一种存活因子，可保护肝脏免受各种损伤，而ALR表达下调可加速小鼠MASLD的进展。CD36是一种长链游离脂肪酸转运蛋白，在MASLD的发病过程中起着至关重要的作用。本研究发现，经过油酸/棕榈酸处理后，CD36在alr转染（ALR-Tx）的HepG2细胞中表达水平显著降低，在shRNA-ALR （shALR）细胞中表达水平上调。此外，在高脂肪饮食中杂合子Gfer敲除（Gfer+/-）的MASLD模型小鼠以及肝脏特异性Gfer敲除（Gfer- cko）的模型小鼠中，CD36的表达显著升高。在shALR HepG2细胞中，脂质摄取增加，脂质吞噬被抑制，而在ALR-Tx HepG2细胞中，脂质摄取减少，脂质吞噬被促进。在shALR细胞中，siRNA-CD36下调CD36表达后，脂质积累明显减少，刺激脂质吞噬。ATF3（激活转录因子3）mRNA和蛋白表达在ALR-Tx细胞和shALR细胞中分别显著升高和降低，敲低ATF3后可诱导CD36表达。因此，我们的研究结果表明，ALR的缺乏在一定程度上通过ATF3-CD36轴加剧了MASLD中的脂质沉积。

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引用次数: 0

Timeless prevents senescence-associated phenotypes and enhances DNA repair to promote esophageal cancer cell growth 永恒预防衰老相关表型和增强DNA修复促进食管癌细胞生长。

IF 3.5 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-11-13 DOI: 10.1016/j.yexcr.2025.114828

Chiaki Noguchi , Kalisse I. Horne , Tylor Brewster , Alyssa Duffy , Jeel P. Shah , Amber Theriault , Olivia El Naggar , Soumya Vavilala , Shivani Sheth , Eishi Noguchi

Esophageal squamous cell carcinoma (ESCC) is one of the deadliest forms of squamous cell carcinoma, comprising approximately 90 % of all esophageal cancer cases. We previously demonstrated that the Fanconi anemia DNA repair (FA) pathway mitigates replication stress to preserve self-renewal capacity of esophageal cancer cells, highlighting the critical role of minimizing replication stress in esophageal cancer proliferation. In this study, to further explore the role of replication stress in esophageal cancer growth, we investigated the function of Timeless, a key subunit of the replication fork protection complex essential for preventing replication stress. Our findings reveal that Timeless is upregulated in esophageal cancer cells, and its depletion increases sensitivity to DNA-damaging agents, inducing cellular senescence in esophageal keratinocytes. Timeless depletion also elevates the DNA damage response while reducing the expression of DNA repair proteins associated with the FA pathway and homologous recombination. Furthermore, the loss of Timeless impairs colony-forming ability in soft agar and diminishes the self-renewal capacity required to form 3D organoids. These results suggest that Timeless plays a critical role in facilitating DNA repair and esophageal cancer progression and may represent a promising target for developing effective therapeutic strategies to treat esophageal cancers.

食管鳞状细胞癌（ESCC）是最致命的鳞状细胞癌之一，约占所有食管癌病例的90%。我们之前证明了Fanconi贫血DNA修复（FA）途径减轻复制应激以保持食管癌细胞的自我更新能力，强调了最小化复制应激在食管癌增殖中的关键作用。在这项研究中，为了进一步探讨复制应激在食管癌生长中的作用，我们研究了复制叉保护复合体的一个关键亚基Timeless的功能，该亚基是防止复制应激所必需的。我们的研究结果表明，在食管癌细胞中，Timeless表达上调，其缺失增加了对dna损伤剂的敏感性，从而诱导食管角化细胞衰老。长期耗竭也会提高DNA损伤反应，同时降低与FA通路和同源重组相关的DNA修复蛋白的表达。此外，Timeless的缺失削弱了软琼脂中的集落形成能力，并降低了形成三维类器官所需的自我更新能力。这些结果表明，Timeless在促进DNA修复和食管癌进展中起着关键作用，可能是开发有效治疗食管癌策略的有希望的靶点。

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引用次数: 0

Targeting LncRNA MEG3 to modulate ER stress and autophagy: A CRISPR/Cas9-based strategy in AKI-to-CKD transition 靶向LncRNA MEG3调节内质网应激和自噬：一种基于CRISPR/ cas9的aki向ckd过渡策略

IF 3.5 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-11-11 DOI: 10.1016/j.yexcr.2025.114826

Bhupendra Puri , Syamantak Majumder , Anil Bhanudas Gaikwad

Acute kidney injury (AKI) to chronic kidney disease (CKD) transition is a progressive, long-term kidney dysfunction driven by complex pathophysiological processes, including persistent endoplasmic reticulum (ER) stress and impaired autophagy, contributing to fibrosis. Long non-coding RNA (lncRNA) maternally expressed gene 3 (MEG3) has been implicated in endoplasmic reticulum (ER) stress and autophagy in several diseases, but its role in kidney injury and fibrosis during AKI-to-CKD transition remains unclear. Our previous transcriptomic analysis revealed that MEG3 is dysregulated during this transition, prompting us to explore its functional role. In this study, we investigated the function of MEG3 in ER stress–autophagy crosstalk during the AKI-to-CKD transition. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR/Cas9)-mediated MEG3 knockout in NRK-52E cells was confirmed by T7 endonuclease assay, quantitative real-time polymerase chain reaction (qRT-PCR), and fluorescence in-situ hybridization (FISH) assay. Functionally, MEG3 knockout markedly attenuated ER stress and apoptosis, as shown by reduced expression of BiP, CHOP, ATF6, ATF4, p-PERK, p-eIF2α, along with restoration of the Bax/Bcl-2 balance. Autophagy activity was restored, with increased Beclin-1 and LC3B expression and decreased p62 accumulation. Furthermore, fibrotic progression was reduced, as indicated by lower levels of fibronectin and collagen I. Notably, tauroursodeoxycholic acid (TUDCA, 400 μM) acted synergistically with MEG3 knockout, further suppressing ER stress and fibrosis markers compared to either treatment alone. These findings demonstrate that MEG3 promotes maladaptive ER stress and impaired autophagy in tubular epithelial cells, driving AKI-to-CKD transition. Targeting MEG3 through CRISPR-based strategies or in combination with TUDCA may represent a promising therapeutic strategy to mitigate fibrosis and slow disease progression.

急性肾损伤（AKI）向慢性肾脏疾病（CKD）的转变是由复杂的病理生理过程驱动的进行性、长期肾功能障碍，包括持续的内质网（ER）应激和自噬受损，导致纤维化。长链非编码RNA （lncRNA）母系表达基因3 （MEG3）与多种疾病的内质网（ER）应激和自噬有关，但其在aki向ckd过渡期间肾损伤和纤维化中的作用尚不清楚。我们之前的转录组学分析显示，MEG3在这一转变过程中是失调的，这促使我们探索其功能作用。在这项研究中，我们研究了MEG3在aki到ckd转变过程中内质网应激-自噬串扰中的功能。T7核酸内切酶（T7）、定量逆转录聚合酶链反应（qRT-PCR）和荧光原位杂交（FISH）实验证实了聚集规律间隔短回文重复（CRISPR/Cas9）介导的MEG3基因敲除在NRK-52E细胞中的作用。功能上，MEG3敲除显著减轻内质网应激和细胞凋亡，如BiP、CHOP、ATF6、ATF4、p-PERK、p-eIF2α的表达降低，以及Bax/Bcl-2平衡的恢复。自噬活性恢复，Beclin-1和LC3B表达增加，p62积累减少。此外，纤维连接蛋白和胶原蛋白水平降低表明，纤维化进展减少。值得注意的是，与单独治疗相比，tauroursodeoxycholic acid （TUDCA, 400 μM）与MEG3敲除协同作用，进一步抑制ER应激和纤维化标志物。这些发现表明，MEG3促进小管上皮细胞的内质网络应激不良和自噬损伤，推动aki向ckd过渡。通过基于crispr的策略靶向MEG3或与TUDCA联合可能是缓解纤维化和减缓疾病进展的有希望的治疗策略。

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引用次数: 0

Gastrodin produces therapeutic effects against preeclampsia by activating Wnt 3a signaling and inhibiting ferroptosis 天麻素通过激活Wnt 3a信号和抑制铁下垂产生对子痫前期的治疗作用。

IF 3.5 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-11-10 DOI: 10.1016/j.yexcr.2025.114827

Qiang Xu , Ge Xuan , Chongying Zhu , Bingquan Zhu , Zihao An , Meiyuan Jin , Chao Tang

Preeclampsia (PE) is a multisystem disorder with increased maternal and perinatal mortality and morbidity while the treatment of PE remains largely unknown in clinic. Therefore, it is necessary to find more effective therapeutic methods of PE. We report herein that, Tianma Gouteng Decoction (TGD) generates protective effects against PE by activating Wnt 3a/β-catenin signaling and simultaneously suppressing ferroptosis in placental trophoblast cells. Particularly, Gastrodin (Gtd), the active component in TGD, plays a major role in protection against PE. Mechanistically, Gtd up-regulates the Wnt 3a/β-catenin signaling activity by inducing Wnt 3a mRNA expression, resulting in the increased expression of β-catenin-controlled target genes. On the other hand, Gtd-triggered Wnt activation obviously exerts negative effects on ferroptosis by promoting expression levels of the anti-ferroptosis proteins accompanied by the down-regulated reactive oxygen species (ROS) production and total iron content but the up-regulated L-Glutathione (GSH) levels. Consistently, Gtd-administration reveals apparent anti-hypertensive effects in a PE-like mouse model with diminished ferroptosis, whereas deactivation of β-catenin by administration with the specific antagonist ICG001 disrupts the protective effects derived from Gtd. Therefore, our results provide an innovative basis for the role of Gtd as a new therapy for PE.

先兆子痫（PE）是一种多系统疾病，增加了孕产妇和围产期死亡率和发病率，而PE的治疗方法在临床上仍然很大程度上未知。因此，有必要寻找更有效的PE治疗方法。我们在此报道，天麻钩藤汤（TGD）通过激活Wnt 3a/β-catenin信号通路，同时抑制胎盘滋养细胞铁凋亡，从而对PE产生保护作用。特别是天麻素（Gtd），天麻素是TGD的活性成分，在PE的保护中起着重要作用。机制上，Gtd通过诱导Wnt 3a mRNA表达上调Wnt 3a/β-catenin信号转导活性，导致β-catenin控制靶基因表达增加。另一方面，gtd触发的Wnt激活对铁下垂有明显的负面影响，通过促进抗铁下垂蛋白的表达水平，同时降低活性氧（ROS）的产生和总铁含量，而上调l -谷胱甘肽（GSH）水平。与此一致的是，Gtd给药在pe样小鼠模型中显示出明显的抗高血压作用，并减少了铁凋亡，而与特异性拮抗剂ICG001一起给药会使β-catenin失活，从而破坏了Gtd的保护作用。因此，我们的研究结果为Gtd作为PE新疗法的作用提供了创新基础。

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