Biochimica et biophysica acta. Molecular cell research最新文献_第3页

Assessing the involvement of tumor-secreted factors in the inhibition of muscle differentiation 评估肿瘤分泌因子在肌肉分化抑制中的作用

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-12-04 DOI: 10.1016/j.bbamcr.2025.120093

Miranda van der Ende , Xiaolin Li , Mieke Poland , Fleur Jansen , Jocelijn Meijerink , Jaap Keijer , Renger F. Witkamp , Sander Grefte , Klaske van Norren

Cancer cachexia is a multifactorial syndrome characterized by involuntary and pathological weight loss, predominantly caused by muscle wasting. While tumors can elicit detrimental effects on skeletal muscle function, the contribution of specific tumor-derived mediators remains elusive. To explore this, we investigated the impact of conditioned media (CM) from four cachexia-inducing tumor cell lines (KPC, 4662, LLC, and C26) on muscle differentiation using C2C12 cells. Creatine kinase (CK) activity was measured as an indicator of muscle wasting, and global gene expression changes in C2C12 cells were analyzed via RNA sequencing. Cytokine profiling of the CM identified 111 immune factors, and mimic combinations of the most abundant cytokines from KPC CM were tested for their effects on CK activity. Additionally, the involvement of tumor-derived PGE2 was assessed via CRISPR/Cas9-mediated knockout of the Ptgs2 gene in KPC cells. CM from all tumor cell lines significantly reduced CK activity in C2C12 cells, consistent with downregulation of CKm gene expression. Global gene expression profiles revealed upregulation of immune-related pathways in C2C12 cells exposed to KPC CM. However, mixtures of the 14 most abundant cytokines in CM had minimal effects on CK activity, and tumor-derived PGE2 showed no significant effect on CK activity or muscle cell differentiation. These findings suggest that the observed muscle-wasting effects of cachexia-inducing tumor cells cannot be replicated by the most abundant cytokines present in CM alone, highlighting the need for further research to identify the key tumor-derived factors responsible for cancer-induced muscle wasting.

癌症恶病质是一种多因素综合征，以不自主和病理性体重减轻为特征，主要由肌肉萎缩引起。虽然肿瘤可引起对骨骼肌功能的有害影响，但特异性肿瘤衍生介质的贡献仍然难以捉摸。为了探讨这一点，我们研究了来自四种恶病质诱导肿瘤细胞系（KPC、4662、LLC和C26）的条件培养基（CM）对C2C12细胞肌肉分化的影响。测定肌酸激酶（CK）活性作为肌肉萎缩的指标，并通过RNA测序分析C2C12细胞整体基因表达变化。细胞因子分析鉴定了111个免疫因子，并测试了KPC CM中最丰富的细胞因子模拟组合对CK活性的影响。此外，通过CRISPR/ cas9介导的敲除KPC细胞中的Ptgs2基因来评估肿瘤源性PGE2的参与。所有肿瘤细胞系的CM均显著降低C2C12细胞的CK活性，与CKm基因表达下调一致。全球基因表达谱显示，暴露于KPC CM的C2C12细胞中免疫相关通路上调。然而，CM中14种最丰富的细胞因子的混合物对CK活性的影响很小，肿瘤来源的PGE2对CK活性或肌肉细胞分化没有显著影响。这些发现表明，观察到的恶病质诱导肿瘤细胞的肌肉萎缩效应不能被CM中最丰富的细胞因子所复制，这突出了进一步研究确定癌症诱导肌肉萎缩的关键肿瘤来源因素的必要性。

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引用次数: 0

Heat shock protein 70 regulates m6A modification in response to heat shock in esophageal squamous cell carcinoma. 热休克蛋白70在食管鳞状细胞癌热休克反应中调控m6A修饰。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-12-01 Epub Date: 2025-07-24 DOI: 10.1016/j.bbamcr.2025.120027

Bin Du, Jia Wang, Jun Ma, Pu Wang

Heat shock has been known to induce hyperplasia in esophageal epithelial cells. It is widely considered as a crucial risk factor in the initiation and development of esophageal squamous cell carcinoma (ESCC), yet our understanding of the underlying mechanisms remains limited. The m6A modification of mRNA plays a role in mediating several cellular processes and is critical during cell stress. Our study revealed that inhibiting of m6A 'writer' components of ESCC cells exhibit higher death rates and slower recovery after heat shock. After normalization using mRNA expression profiles, 91.08 % of significantly changed m6A modifications aligned with corresponding mRNA abundance changes, with no evidence of over-modification, while the increase in m6A modification of 8.92 % of heat-shock associated genes far exceeded the increase in mRNA (hyper - m6A modification), and A/U rich motifs were commonly observed in the 3'UTR of these gene. Inside the nucleus, the binding of HSP70s in m6A writer complex promote the hyper - m6A modification in specific mRNAs after heat shock. The stronger nuclear localization of HSP70 in ESCC tissues correlates with a poor prognosis for the patients. In conclusion, our research revealed that the nuclear HSP70 protein could bind to the METTL3/14 writer complex and regulate mRNA's m6A modification. Our results provide a new perspective for research into how HSP70 protein regulates mRNA stability and suggests a new direction for the comprehensive prevention and treatment of ESCC.

已知热休克可诱导食管上皮细胞增生。它被广泛认为是食管鳞状细胞癌（ESCC）发生和发展的关键危险因素，但我们对其潜在机制的了解仍然有限。mRNA的m6A修饰在介导几种细胞过程中起作用，在细胞应激过程中起关键作用。我们的研究表明，严重缺乏m6A ‘writer’成分的ESCC细胞在热休克后表现出更高的凋亡率和更慢的恢复速度。对mRNA表达谱进行归一化后，91.07 %的m6A修饰显著变化与相应的mRNA丰度变化一致，没有过度修饰的证据，而8.92 %的热休克相关基因m6A修饰的增加远远超过mRNA的增加（超m6A修饰），并且在这些基因的3'UTR中普遍观察到A/U富基序。在细胞核内，hsp70在m6A写入复合体中的结合促进了热休克后特定mrna的超m6A修饰。ESCC组织中HSP70的核定位越强，患者预后越差。综上所述，我们的研究揭示了核HSP70蛋白可以结合METTL3/14复合体并调节mRNA的m6A修饰。本研究结果为研究HSP70蛋白调控mRNA稳定性提供了新的视角，为ESCC的综合防治提供了新的方向。

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引用次数: 0

Cancer-associated fibroblasts secreting IL-6 inhibit the cisplatin and docetaxel killing effect in lung squamous cell carcinoma. 分泌IL-6的癌相关成纤维细胞抑制顺铂和多西他赛对肺鳞癌的杀伤作用。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-12-01 Epub Date: 2025-07-31 DOI: 10.1016/j.bbamcr.2025.120029

Xu Zhu, Long Li, Xuanyin Wang, Ying Zhang, Zeyang Yang, Jiaming Ren, Lu Wang, Xianling Zeng, Jing Xiao, Siyu Wang, Peiying Pan, Jian Zhang, Xiaojun Du, Tao Wang, Langbo Liu, Xiaolin Shu, Qiaoling Zhang, Jiangwei Wu, Siyuan Yang, Xinlei Liu, Zhu Zeng, Jieheng Wu

Chemoresistance remains a major obstacle in the treatment of lung squamous cell carcinoma (LUSC), often leading to suboptimal clinical outcomes. Among the key contributors to this resistance are cancer-associated fibroblasts (CAFs), which are increasingly recognized for their tumor-supportive roles. Despite this, the molecular pathways through which CAFs promote chemoresistance in LUSC are not fully elucidated. This study found that CAFs-derived interleukin-6 (IL-6) upregulated the expression of Specificity Protein 1 (SP1) and the ATP-binding cassette transporter B7 (ABCB7) in LUSC cells exposed to cisplatin and docetaxel. In vitro assays showed a marked decrease in apoptosis in tumor cells co-cultured with CAFs. Consistent with these findings, in vivo xenograft models demonstrated that IL-6-producing CAFs reduced the antitumor efficacy of both chemotherapeutic agents. Elevated serum IL-6 levels also emerged as a potential indicator of poor response to chemotherapy. Our findings suggest that IL-6 secreted by CAFs impairs the cytotoxic effects of cisplatin and docetaxel in LUSC, partly through activation of the PI3K/AKT/NF-κB signaling axis. Targeting this IL-6-mediated pathway may offer a promising strategy to overcome chemoresistance and enhance therapeutic outcomes in patients with LUSC.

化疗耐药仍然是肺鳞状细胞癌（LUSC）治疗的主要障碍，经常导致不理想的临床结果。这种耐药的关键因素是癌症相关成纤维细胞（CAFs），其肿瘤支持作用日益得到认可。尽管如此，CAFs促进LUSC化学耐药的分子途径尚未完全阐明。本研究发现，在暴露于顺铂和多西紫杉醇的LUSC细胞中，cafs来源的白介素-6 （IL-6）上调特异性蛋白1 （SP1）和atp结合盒转运蛋白B7 （ABCB7）的表达。体外实验显示，与CAFs共培养的肿瘤细胞凋亡明显减少。与这些发现一致，体内异种移植模型表明，产生il -6的CAFs降低了两种化疗药物的抗肿瘤疗效。血清IL-6水平升高也成为化疗反应不良的潜在指标。我们的研究结果表明，由CAFs分泌的IL-6损害顺铂和多西紫杉醇在LUSC中的细胞毒性作用，部分通过激活PI3K/AKT/NF-κB信号轴。靶向这种il -6介导的途径可能为克服LUSC患者的化疗耐药和提高治疗效果提供了一种有希望的策略。

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引用次数: 0

SPRED2 controls the severity of cisplatin-induced acute kidney injury by inhibiting ERK activation and TNFα production in mice SPRED2通过抑制小鼠ERK活化和TNFα产生来控制顺铂诱导的急性肾损伤的严重程度

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-11-30 DOI: 10.1016/j.bbamcr.2025.120091

Xu Yang , Jiali He , Tong Gao , Masayoshi Fujisawa , Toshiaki Ohara , Steven L. Kunkel , Teizo Yoshimura , Akihiro Matsukawa

Cisplatin is an effective chemotherapeutic agent used to treat solid tumors, but its clinical use is limited by acute kidney injury (AKI), in which ERK signaling plays a crucial role. Here, we investigated whether Sprouty-related EVH1 domain-containing protein 2 (SPRED2), an endogenous inhibitor of the Ras/Raf/ERK pathway, protects against cisplatin-induced AKI. Spred2^−/− mice showed more severe renal injury and stronger ERK activation than wild-type (WT) mice, whereas pretreatment with the MEK inhibitor U0126 markedly attenuated the injury. In HK-2 cells (proximal tubular cells), SPRED2 knockdown enhanced cisplatin-induced apoptosis and caspase-3 activation, accompanied by decreased Bcl-2 expression. Spred2^−/− kidneys displayed increased macrophage infiltration and elevated Tnfα, Il1b, and Ccl2 expression. Neutralization of TNFα with anti-TNFα antibody ameliorated renal injury and reduced the levels of Il1b and Ccl2 mRNA in Spred2^−/− mice. In vitro, TNFα slightly decreased the viability of control and SPRED2 knockdown HK-2 cells without cisplatin treatment, but the decreased viability was augmented in SPRED2 knockdown cells by cisplatin. Immunohistochemistry revealed that macrophages were the predominant TNFα-positive cell population. Bone marrow–derived macrophages from Spred2^−/− mice produced higher levels of TNFα in response to cisplatin compared with control cells, and this increase was markedly suppressed by U0126.

These findings indicate that endogenous SPRED2 protects kidneys from cisplatin-induced AKI by limiting ERK activation, tubular apoptosis, and TNFα-mediated inflammation.

顺铂是一种用于治疗实体瘤的有效化疗药物，但其临床应用受到急性肾损伤（AKI）的限制，其中ERK信号在其中起着至关重要的作用。在这里，我们研究了sprouty相关的EVH1结构域蛋白2 (SPRED2)，一种Ras/Raf/ERK途径的内源性抑制剂，是否对顺铂诱导的AKI有保护作用。与野生型（WT）小鼠相比，Spred2−/−小鼠表现出更严重的肾损伤和更强的ERK激活，而MEK抑制剂U0126预处理可显著减轻损伤。在HK-2细胞（近端小管细胞）中，SPRED2敲低可增强顺铂诱导的凋亡和caspase-3激活，并伴有Bcl-2表达降低。Spred2−/−肾脏显示巨噬细胞浸润增加，Tnfα、Il1b和Ccl2表达升高。用抗tnf - α抗体中和tnf - α可改善Spred2−/−小鼠的肾损伤，并降低il - 1b和Ccl2 mRNA的水平。体外实验中，未经顺铂治疗的对照组和SPRED2敲低的HK-2细胞的活力略有下降，但在SPRED2敲低的细胞中，顺铂增强了这种下降的活力。免疫组化结果显示，tnf α阳性细胞群以巨噬细胞为主。与对照细胞相比，Spred2 - / -小鼠骨髓源性巨噬细胞对顺铂产生更高水平的tnf - α，而U0126明显抑制了这种增加。这些发现表明，内源性SPRED2通过限制ERK激活、肾小管凋亡和tnf α介导的炎症，保护肾脏免受顺铂诱导的AKI。

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引用次数: 0

Structural diversity of full-length human αvβ3 integrin revealed by cryo-EM 低温电镜分析人αvβ3整联素的结构多样性。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-11-29 DOI: 10.1016/j.bbamcr.2025.120089

Cang Wu , Yuanzhu Gao , Weiyan Wang , Zhoufang Li , Qing Shu , Xi Zhang

Integrins are essential transmembrane receptors that orchestrate cell adhesion, migration, and survival, and have emerged as promising therapeutic targets for cancer, fibrosis, and autoimmune diseases. However, most integrin-targeted drugs have failed in clinical trials due to limited efficacy and unexpected off-target effects, reflecting an incomplete understanding of integrin conformational regulation. Here, we present a series of high-resolution cryo-EM structures of human integrin αvβ3 in both apo and ligand-bound states by collecting a large amount of data. Consequently, we resolved six conformations of integrin in the apo state, five of which were previously uncharacterized, along with five distinct ligand-bound states, thereby revealing a continuum of conformational transitions underlying integrin activation. Notably, CWHM-12 enables the simultaneous coexistence of integrin in closing and opening inhibited states, revealing a mechanism that differs fundamentally from that of conventional RGD peptide-based inhibitors. Then, our study provides a structural framework for understanding integrin activation diversity and lays the foundation for rational design of next-generation inhibitors with improved precision and reduced off-target effects.

整合素是协调细胞粘附、迁移和存活的重要跨膜受体，已成为癌症、纤维化和自身免疫性疾病的有希望的治疗靶点。然而，大多数整合素靶向药物由于疗效有限和意想不到的脱靶效应而在临床试验中失败，反映了对整合素构象调控的不完全理解。在此，我们通过收集大量数据，获得了一系列载脂蛋白和配体结合态的人整合素αvβ3的高分辨率冷冻电镜结构。因此，我们在载脂蛋白状态下解析了整合素的六种构象，其中五种以前未被表征，以及五种不同的配体结合状态，从而揭示了整合素激活下的连续构象转变。值得注意的是，CWHM-12使整合素同时处于关闭和打开抑制状态，揭示了一种与传统RGD肽抑制剂根本不同的机制。因此，我们的研究为理解整合素激活多样性提供了一个结构框架，并为合理设计具有更高精度和减少脱靶效应的下一代抑制剂奠定了基础。

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引用次数: 0

Metabolic reprogramming is implicated in the differential response of the CAL-1 plasmacytoid dendritic cell line to autophagy inhibitors 代谢重编程涉及CAL-1浆细胞样树突状细胞系对自噬抑制剂的差异反应。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-11-27 DOI: 10.1016/j.bbamcr.2025.120090

Carlota Ramalhinho , Paulo Antas , Philippe Pierre , Iola F. Duarte , Catarina R. Almeida

Plasmacytoid dendritic cells (pDCs) produce large amounts of type I Interferons (IFN) and pro-inflammatory cytokines, playing crucial roles in antiviral and anticancer immunity as well as in autoimmune diseases. Understanding the mechanisms that regulate pDC function is therefore essential. Autophagy, a process responsible for recycling intracellular components, influences cellular metabolism and modulates immune responses. Here, we used the human CAL-1 pDC cell line, a validated model for primary pDCs, to investigate the functional impact of autophagy inhibition and the contribution of metabolism to these effects. CAL-1 cells were treated with two autophagy inhibitors, Spautin-1 and VPS34-IN1, and cytokine production was assessed by RT-qPCR and ELISA, while cellular metabolism was analysed by untargeted metabolomics of cell extracts and of medium supernatants. VPS34-IN1, but not Spautin-1, induced robust expression of IFN-β and TNF-α. The two inhibitors also elicited distinct metabolic responses: Spautin-1 enhanced glycolysis, promoted an anabolic phenotype with increased utilization of amino acids, and upregulated mTOR signaling. In contrast, VPS34-IN1 decreased glycolysis, increased intracellular amino acids, reduced TCA intermediates, and induced energy stress, reflected by an increased AMP/(ADP + ATP) ratio and decreased NAD⁺. These changes were consistent with AMPK activation, and pharmacological inhibition of AMPK with dorsomorphin (compound C) abolished cytokine production in VPS34-IN1-treated cells. Together, these results indicate that differential cytokine responses to autophagy inhibition are driven primarily by metabolic rewiring rather than autophagic flux per se, highlighting the interplay between metabolism, mitochondrial ROS, and signaling pathways in pDC activation.

浆细胞样树突状细胞（pDCs）产生大量I型干扰素（IFN）和促炎细胞因子，在抗病毒和抗癌免疫以及自身免疫性疾病中起着至关重要的作用。因此，了解调控pDC功能的机制至关重要。自噬是一种负责细胞内成分循环的过程，影响细胞代谢并调节免疫反应。在这里，我们使用人CAL-1 pDC细胞系（一种验证过的原代pDC模型）来研究自噬抑制对功能的影响以及代谢对这些影响的贡献。用两种自噬抑制剂Spautin-1和VPS34-IN1处理CAL-1细胞，通过RT-qPCR和ELISA检测细胞因子的产生，通过细胞提取物和培养基上清液的非靶向代谢组学分析细胞代谢。VPS34-IN1诱导IFN-β和TNF-α的强烈表达，而Spautin-1不诱导。这两种抑制剂也引发了不同的代谢反应：Spautin-1增强糖酵解，促进合成代谢表型，增加氨基酸的利用，上调mTOR信号。相反，VPS34-IN1降低糖酵解，增加细胞内氨基酸，减少TCA中间体，诱导能量应激，表现为AMP/（ADP + ATP）比值升高和NAD+降低。这些变化与AMPK的激活一致，用dorsomorphin（化合物C）抑制AMPK可消除vps34 - in1处理的细胞中细胞因子的产生。总之，这些结果表明，细胞因子对自噬抑制的差异反应主要是由代谢重布线驱动的，而不是自噬通量本身，这突出了代谢、线粒体ROS和pDC激活信号通路之间的相互作用。

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引用次数: 0

Transgenic overexpression of miR-486 and sAnk1.5 does not alter glucose handling in mice 转基因过表达miR-486和sAnk1.5不会改变小鼠的葡萄糖处理。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-11-17 DOI: 10.1016/j.bbamcr.2025.120087

S. Buonocore , F. Fiore , E.M. Rubino , M.R. Catallo , L. Raucci , A. Laurino , D. Rossi , E. Pierantozzi , V. Sorrentino

Recent studies have shown that the C allele of SNP rs508419 is associated with susceptibility to Type 2 diabetes (T2D). This SNP lies within the muscle-specific P2 promoter of human ANK1, which drives transcription of the sAnk1.5 isoform and miR-486. The C allele increases P2 promoter activity, resulting to higher levels of sAnk1.5 transcript and protein in striated muscles.

We now present the first evidence that in skeletal muscle of individuals homozygous for the rs508419 C allele, also the hsa-miR-486-5p is transcribed at higher levels. This raises the question of whether T2D susceptibility may be associated with simultaneous overexpression of miR-486-5p and sAnk1.5. To test this hypothesis, we generated and characterized double transgenic (D-Tg) mice that selectively overexpress both mmu-miR-486-5p and sAnk1.5 in skeletal muscle tissue.

Analysis of sAnk1.5 and miR-486-5p expression in D-Tg mouse showed that despite both transgenes were significantly upregulated, a discrepancy between sAnk1.5 mRNA and protein levels was observed, suggesting that sAnk1.5 protein levels are further regulated by post-translational mechanism. D-Tg mice were monitored from 2 to 12 months of age to assess body weight, fat and lean mass, blood glucose levels under fasting conditions, as well as during intraperitoneal glucose tolerance tests and insulin tolerance tests, performed under either standard or high fat diet conditions. No differences were observed between D-Tg and age-matched wild type control mice for any of the parameters tested, indicating that the link between rs508419 and susceptibility to T2D cannot be ascribed to increased expression of miR-486-5p and sAnk1.5 in skeletal muscle.

最近的研究表明SNP rs508419的C等位基因与2型糖尿病（T2D）易感性相关。该SNP位于人类ANK1的肌肉特异性P2启动子内，其驱动sAnk1.5亚型和miR-486的转录。C等位基因增加P2启动子活性，导致横纹肌中sAnk1.5转录物和蛋白水平升高。我们现在提出了第一个证据，即在rs508419 C等位基因纯合的个体的骨骼肌中，hsa-miR-486-5p的转录水平也更高。这就提出了T2D易感性是否可能与miR-486-5p和sAnk1.5同时过表达有关的问题。为了验证这一假设，我们产生并表征了双转基因（D-Tg）小鼠，这些小鼠在骨骼肌组织中选择性地过表达mmu-miR-486-5p和sAnk1.5。对D-Tg小鼠中sAnk1.5和miR-486-5p表达的分析显示，尽管两种转基因均显著上调，但sAnk1.5 mRNA和蛋白水平存在差异，表明sAnk1.5蛋白水平进一步受到翻译后机制的调控。D-Tg小鼠在2至12 月龄期间进行监测，以评估空腹条件下的体重、脂肪和瘦质量、血糖水平，以及在标准或高脂肪饮食条件下进行的腹腔葡萄糖耐量试验和胰岛素耐量试验。在D-Tg和年龄匹配的野生型对照小鼠之间，没有观察到任何参数的差异，这表明rs508419与T2D易感性之间的联系不能归因于骨骼肌中miR-486-5p和sAnk1.5的表达增加。

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引用次数: 0

Anti-tumor response by targeting glucose metabolism and depletion of membrane cholesterol in breast cancer and melanoma cells 针对乳腺癌和黑色素瘤细胞中葡萄糖代谢和膜胆固醇消耗的抗肿瘤反应。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-11-16 DOI: 10.1016/j.bbamcr.2025.120088

Himanshi Yaduvanshi, Bhavana Deshmukh, Abhijeet Singh, Shyamananda Singh Mayengbam, Manoj Kumar Bhat

Enhanced lactate production and cholesterol accumulation in cancer cells are often correlated with poor prognosis and the development of resistance to therapies. This study investigates a unique dual-pronged approach for abrogating tumor development by combining methyl-β-cyclodextrin (MCD)-mediated cholesterol depletion with sodium oxamate (OXA)-induced inhibition of lactate dehydrogenase A.

The cytotoxic effects of MCD and OXA were validated by MTT and long-term colony formation assay. Cell cycle arrest and apoptosis were assessed via flow cytometry. The involvement of key signaling intermediates in proliferative pathways was evaluated by immunoblotting. The Seahorse analyzer was used to measure the real-time metabolic flux in mouse cancer cells. In vivo, studies and immunophenotyping were performed to investigate the impact of MCD+OXA treatment on tumor growth and immune cell infiltration in tumor and periphery organs. The results indicate that concurrent administration of a low-dose MCD and OXA elicited a synergistic cytotoxic effect and retarded tumor progression while being non-toxic to vital organs. Mechanistically, this combination downregulated ERK/AKT signaling, induced apoptosis, and suppressed cellular metabolism at the glycolysis/OXPHOS level, including ATP production. Interestingly, it promoted the infiltration of effector Th1 cells and B cells, while reducing the presence of PMN-MDSCs in the tumor microenvironment and peripheral organs. These findings suggest that the combination of MCD and OXA provides a dual-targeted approach by disrupting cholesterol as well as lactate metabolism and eliciting an anti-tumor immune response, leading to a reduction in the growth of tumor cells. This pre-clinical strategy demonstrates potential advantages over single-agent treatment, which warrants further investigations for its therapeutic implications.

Summary blurb

In pre-clinical investigations, co-targeting membrane cholesterol, and lactate dehydrogenase A with methyl-β-cyclodextrin and sodium oxamate inhibits cell growth and restricts breast cancer and melanoma tumor progression.

癌细胞中乳酸生成和胆固醇积累的增加通常与预后不良和对治疗产生耐药性有关。本研究探讨了一种独特的双管齐下的方法，通过结合甲基β-环糊精（MCD）介导的胆固醇消耗和草酸钠（OXA）诱导的乳酸脱氢酶a的抑制来消除肿瘤的发展。MCD和OXA的细胞毒性作用通过MTT和长期集落形成实验得到验证。流式细胞术检测细胞周期阻滞和凋亡情况。通过免疫印迹法评估关键信号中间体在增殖途径中的作用。采用海马分析仪实时测定小鼠癌细胞的代谢通量。在体内，我们通过研究和免疫分型来研究MCD+OXA治疗对肿瘤生长和肿瘤及周围器官免疫细胞浸润的影响。结果表明，同时给予低剂量MCD和OXA可引起协同细胞毒作用，延缓肿瘤进展，同时对重要器官无毒。在机制上，该组合下调ERK/AKT信号，诱导细胞凋亡，并抑制糖酵解/OXPHOS水平的细胞代谢，包括ATP的产生。有趣的是，它促进了效应Th1细胞和B细胞的浸润，同时减少了PMN-MDSCs在肿瘤微环境和外周器官中的存在。这些发现表明，MCD和OXA的结合提供了一种双靶向方法，通过破坏胆固醇和乳酸代谢，引发抗肿瘤免疫反应，导致肿瘤细胞生长减少。这种临床前策略显示了比单药治疗的潜在优势，值得进一步研究其治疗意义。摘要：在临床前研究中，甲基β-环糊精和草酸钠共同靶向膜胆固醇和乳酸脱氢酶A抑制细胞生长并限制乳腺癌和黑色素瘤肿瘤进展。

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引用次数: 0

Corrigendum to “Activation of ER stress and apoptosis by hydrogen peroxide in HeLa cells: Protective role of mild heat preconditioning at 40 °C” [Biochim. Biophys. Acta Mol. Cell Res., 1813 (2011) 1987–1999. / PMID: 21875624] “过氧化氢在HeLa细胞中内质网应激和凋亡的激活：40°C轻度热预处理的保护作用”的更正[biochem]。Biophys。分子细胞学报，1813(2011)1987-1999。/ pmid: 21875624]。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-11-12 DOI: 10.1016/j.bbamcr.2025.120084

Pragathi Pallepati, Diana A. Averill-Bates

引用次数: 0

Escitalopram disrupts PDK1–Akt signaling in B cells through a structure-dependent mechanism independent of SERT Escitalopram通过独立于SERT的结构依赖机制破坏B细胞中的PDK1-Akt信号。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-11-09 DOI: 10.1016/j.bbamcr.2025.120086

Yu-Lun Tseng , Meng-Ling Chiang

The PI3K–Akt pathway is a central regulator of survival and growth signals across receptor systems, controlled at the membrane-proximal PDK1–Akt signaling node. Although selective serotonin reuptake inhibitors (SSRIs) primarily target the serotonin transporter (SERT), evidence suggests additional effects on intracellular signaling. Escitalopram, a widely used SSRI, was previously shown to inhibit Syk phosphorylation in platelets, raising the possibility of SERT-independent effects on kinase cascades. Here, we investigated whether escitalopram and related compounds interfere with PI3K–Akt signaling in Ramos B lymphoma cells stimulated through the B cell receptor (BCR). Escitalopram and its enantiomer R-citalopram selectively reduced phosphorylation of Akt and its downstream effector FoxO1 while sparing upstream Syk and parallel ERK and PLC–PKC pathways. To localize the site of interference, we combined analyses of PDK1-dependent targets with a PTEN-inhibition model. Escitalopram suppressed PLK1 and S6K1 phosphorylation without altering mTORC2 autophosphorylation, and it also reduced H₂O₂-induced Akt phosphorylation. These complementary results point to a downstream site of interference at the level of PDK1–Akt complex formation, beyond PI3K and PIP3 availability. By contrast, the structurally related analog N-methyl-citalopram showed no effect, reinforcing a structure-dependent mechanism of interference with PDK1–Akt signaling. Functionally, escitalopram enhanced apoptosis in BCR-stimulated cells. Similar inhibition under insulin and SDF-1 stimulation indicates a generalized, receptor-independent effect. These findings reveal a noncanonical, SERT-independent mechanism by which escitalopram disrupts PDK1–Akt activation, with potential relevance to the broader intracellular effects of SSRI treatment.

PI3K-Akt通路是跨受体系统的生存和生长信号的中枢调节因子，受膜-近端PDK1-Akt信号传导节点控制。虽然选择性5 -羟色胺再摄取抑制剂（SSRIs）主要靶向5 -羟色胺转运体（SERT），但有证据表明对细胞内信号传导有其他作用。Escitalopram是一种广泛使用的SSRI，先前被证明可以抑制血小板中的Syk磷酸化，这增加了sert不依赖于激酶级联反应的可能性。在这里，我们研究了艾司西酞普兰及其相关化合物是否通过B细胞受体（BCR）刺激Ramos B淋巴瘤细胞干扰PI3K-Akt信号。艾司西酞普兰及其对映体r -西酞普兰选择性地降低Akt及其下游效应物fox01的磷酸化，同时保留上游的Syk和平行的ERK和PLC-PKC通路。为了定位干扰位点，我们将pdk1依赖靶点的分析与pten抑制模型相结合。艾司西酞普兰抑制PLK1和S6K1磷酸化，但不改变mTORC2的自磷酸化，还能降低h2o2诱导的Akt磷酸化。这些互补的结果指出了PDK1-Akt复合物形成水平的下游干扰位点，超出了PI3K和PIP3的可用性。相比之下，结构相关的类似物n -甲基西酞普兰没有效果，这加强了对PDK1-Akt信号通路干扰的结构依赖机制。在功能上，艾司西酞普兰促进bcr刺激细胞的凋亡。胰岛素和SDF-1刺激下的类似抑制表明了一种广泛的、不依赖于受体的作用。这些发现揭示了escitalopram破坏PDK1-Akt活化的非规范、sert独立机制，可能与SSRI治疗更广泛的细胞内效应相关。

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