Biochimica et Biophysica Acta-Gene Regulatory Mechanisms最新文献_第2页

Transcriptional activation of the PKLR gene by novel erythroid-specific regulatory elements 新的红细胞特异性调控元件对plklr基因转录激活的影响。

IF 3.1 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et Biophysica Acta-Gene Regulatory Mechanisms

Pub Date : 2025-09-16 DOI: 10.1016/j.bbagrm.2025.195116

Yea Woon Kim , Jin Kang , AeRi Kim

The pyruvate kinase L/R (PKLR) gene encodes the L- and R-type isoforms of pyruvate kinase, which catalyze the final step of glycolysis in mammals. The L-type isozyme is mainly found in liver cells, whereas the R-type isozyme is produced specifically in erythroid cells. To investigate the transcriptional activation of the PKLR gene for the R-type isozyme, we analyzed chromatin features—including DNase I sensitivity, histone modifications, and enhancer–promoter interactions—in erythroid K562 and non-erythroid HUVEC cells. Putative regulatory elements, including a promoter and two enhancers, were identified near the PKLR locus in K562 cells. Depletion of H3K4me1 and H3K27ac, histone marks associated with active enhancers, through mutation of histone methyltransferases led to a marked reduction in PKLR transcription, while transcription of a nearby gene remained stable. These regulatory elements were highly occupied by the erythroid-specific transcription factors GATA1 and TAL1. Loss of either factors disrupted local H3K27ac and reduced the recruitment of chromatin-looping factors, resulting in decreased PKLR transcription. Furthermore, CRISPR/Cas9-mediated deletion of the putative regulatory elements significantly diminished PKLR transcription, demonstrating their functional importance. Collectively, these findings highlight the essential role of these regulatory elements in activating PKLR transcription in erythroid cells and emphasize the requirement of erythroid-specific factors for their function.

丙酮酸激酶L/R （PKLR）基因编码丙酮酸激酶的L型和R型同工型，在哺乳动物中催化糖酵解的最后一步。l型同工酶主要存在于肝细胞中，而r型同工酶仅在红细胞中产生。为了研究PKLR基因对r型同工酶的转录激活，我们分析了红系K562和非红系HUVEC细胞的染色质特征，包括DNase I敏感性、组蛋白修饰和增强子-启动子相互作用。在K562细胞的pkr位点附近发现了可能的调控元件，包括一个启动子和两个增强子。与活性增强子相关的组蛋白标记H3K4me1和H3K27ac通过组蛋白甲基转移酶突变导致PKLR转录显著减少，而附近基因的转录保持稳定。这些调控元件被红系特异性转录因子GATA1和TAL1高度占据。任何一个因子的缺失都会破坏局部H3K27ac，减少染色质环因子的募集，导致pkr转录减少。此外，CRISPR/ cas9介导的缺失显著降低了PKLR转录，证明了它们在功能上的重要性。总的来说，这些发现强调了这些调控元件在激活红细胞plklr转录中的重要作用，并强调了红细胞特异性因子对其功能的要求。

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引用次数: 0

RNA-protein interaction techniques - A historical and comparative analysis rna -蛋白质相互作用技术-历史和比较分析。

IF 3.1 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et Biophysica Acta-Gene Regulatory Mechanisms

Pub Date : 2025-09-11 DOI: 10.1016/j.bbagrm.2025.195115

Sourabh Chakrabarty , Sayan Roy , Soumyadip Sarkar , Kusum K. Singh

RNAs intricately orchestrate a myriad of molecular and cellular processes. This includes mRNAs which serve as the essential blueprints for protein synthesis and the non-coding RNAs which act as versatile regulators, influencing gene expression, RNA stability, and even protein function. Over the past few decades, a diverse array of RNA- and protein-centric methodologies have emerged to discern and elucidate the identities of RNA binding proteins (RBPs) and RNAs engaged in RNA-Protein interactions (RPI). The selection of an appropriate method is paramount, given the distinctive advantages and limitations inherent to each technique, in order to effectively address specific biological inquiries. This review spans the spectrum of techniques employed in RNA-Protein interaction studies, ranging from time-honored methods like Electrophoretic Mobility Shift Assay (EMSA) to contemporary approaches such as CRISPR-based RNA-Protein interaction profiling (CBRIP). This review aims not only to list methods but also to provide historical background, discuss the strengths and weaknesses of each approach, and highlight their current applications. Furthermore, it consolidates methodologies tailored to characterize RNA-protein interactions based on their distinct purposes.

rna复杂地协调着无数的分子和细胞过程。这包括作为蛋白质合成基本蓝图的mrna和作为多种调节因子，影响基因表达、RNA稳定性甚至蛋白质功能的非编码RNA。在过去的几十年里，出现了一系列以RNA和蛋白质为中心的方法来辨别和阐明RNA结合蛋白（rbp）和参与RNA-蛋白质相互作用（RPI）的RNA的身份。考虑到每种技术的独特优势和固有局限性，为了有效地处理特定的生物学调查，选择适当的方法是至关重要的。这篇综述涵盖了rna -蛋白质相互作用研究中使用的技术范围，从历史悠久的方法，如电泳迁移转移测定（EMSA）到现代方法，如基于crispr的rna -蛋白质相互作用分析（CBRIP）。本文不仅列出了各种方法，而且还提供了历史背景，讨论了每种方法的优缺点，并强调了它们目前的应用。此外，它还根据rna -蛋白质相互作用的不同目的，整合了专门用于表征它们的方法。

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引用次数: 0

The Catsper3 promoter activity is regulated by the cAMP-Response Element Modulator tau (CREMτ) and the cAMP-Response Element Binding protein 1A (CREBA) transcription factors Catsper3启动子活性受cAMP-Response Element Modulator tau （CREMτ）和cAMP-Response Element Binding protein 1A （CREBA）转录因子调控

IF 3.1 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et Biophysica Acta-Gene Regulatory Mechanisms

Pub Date : 2025-08-25 DOI: 10.1016/j.bbagrm.2025.195114

Diego Eduardo Sánchez-Jasso , Sergio Federico López-Guzmán , Javier Hernández-Sánchez , Rosa María Bermúdez-Cruz , Norma Oviedo

Catsper3 has been shown to be essential for the proper functioning of CatSper calcium channel, sperm hyperactivation and egg fertilization. Although its function and physiological relevance to male fertility have been characterized, nothing is known about its regulation at the transcriptional level. Here, we identified the murine Catsper3 promoter and evaluated the role of CREBA and CREMτ transcription factors (TF) on its regulation. Cloning of a predicted Catsper3 promoter region and further deletion analysis indicates that Catsper3 core promoter is located at −157 to +152 relative to the transcription start site (TSS). Mutation of a TATA box and a Downstream Promoter Element (DPE) did not alter the promoter activity, indicating that this promoter is TATA and DPE independent. Exogenous expression of CREBA and CREMτ increase the Catsper3 promoter activity in the presence of the +268 to +439 region, where two CRE sites were predicted, and mutation of both CRE sites prevents the transactivation by both TFs, suggesting that CREMτ and CREBA may use these sites to promote Catsper3 transcription. Finally, binding of CREBA and CREMτ to both CRE sites was confirmed in vitro by EMSA and ChIP-qPCR assays demonstrated an enrichment of CREBA and CREMτ at the Catsper3 promoter in vivo in the testis but not in liver, indicating its tissue-specific binding. Altogether, these results strongly suggest that Catsper3 gene has a TATA-less, DPE independent promoter encompassed by −157 to +152 region around TSS and that Catsper3 gene expression is directly regulated through two CRE sites by CREBA and CREMτ.

Catsper3已被证明对Catsper3钙通道的正常运作、精子过度激活和卵子受精至关重要。虽然其功能和与男性生育能力的生理相关性已被表征，但其在转录水平上的调控尚不清楚。在此，我们鉴定了小鼠Catsper3启动子，并评估了CREBA和CREMτ转录因子（TF）在其调控中的作用。克隆预测的Catsper3启动子区域和进一步的缺失分析表明，相对于转录起始位点（TSS）， Catsper3核心启动子位于- 157至+152。TATA盒子和下游启动子元件（DPE）的突变没有改变启动子的活性，表明该启动子与TATA和DPE无关。CREBA和CREMτ的外源表达增加了+268至+439区域存在的Catsper3启动子活性，其中预测了两个CRE位点，两个CRE位点的突变阻止了两个tf的反激活，这表明CREMτ和CREBA可能利用这些位点促进Catsper3转录。最后，通过体外EMSA和ChIP-qPCR分析证实CREBA和CREMτ与两个CRE位点的结合表明，体内睾丸中Catsper3启动子处的CREBA和CREMτ富集，而肝脏中没有，表明其组织特异性结合。总之，这些结果强烈表明，Catsper3基因具有一个TATA-less， DPE独立的启动子，包含在TSS周围的- 157至+152区域，并且Catsper3基因的表达由CREBA和CREMτ通过两个CRE位点直接调节。

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引用次数: 0

Expression of CMK2 is controlled by the general stress-response transcriptional factor Msn2 through a single STRE site in budding yeast 在出芽酵母中，CMK2的表达受一般应激反应转录因子Msn2通过一个单一的STRE位点控制。

IF 3.1 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et Biophysica Acta-Gene Regulatory Mechanisms

Pub Date : 2025-08-13 DOI: 10.1016/j.bbagrm.2025.195107

Linghuo Jiang , Yiying Gu , Liudan Wei , Jinrong Feng , Lingxin Pan , Xiufan Liao , Yiping Mo , Chunyu Wei

Mammalian calcium/calmodulin-dependent protein kinase II (CaMKII) is a memory molecule in the brain, and regulates fatty acids and lipid metabolism. As a yeast homolog of CaMKII, Cmk2 is a negative feed-back regulator of calcium signaling in Saccharomyces cerevisiae. Previous systemic studies have shown that 42 transcription factors (TFs) are involved in the control of CMK2 expression under various conditions other than calcium stress, but only one, Crz1, is reported to directly regulate CMK2 expression in response to calcium stress. Here, we show that other 26 TFs, Adr1, Aft2, Cad1, Cst6, Cup2, Dal81, Dal82, Flo8, Gcr2, Haa1, Hfi1, Msn2, Oaf1, Pho4, Ppr1, Rfx1, Rgm1, Rpn4, Sfp1, SIp3, Smp1, Spt10, Stp1, Sum1, Swi4 and Tup1, are involved in the positive control of CMK2 transcription, with 10 of them being calcium stress-specific. In contrast, other four TFs, Hir2, Rph1, Sin3 and Uga3, negatively regulates CMK2 transcription independent of calcium stress. Therefore, multiple TFs directly or indirectly control the transcription of CMK2 in yeast cells. EMSA and ChIP analysis demonstrate that the general stress-responsive Msn2 directly controls the expression of CMK2 through one STRE site, 5′ C₋₁₅₅CCCT 3′, in its promoter. Our genetic study indicates that Crz1 is epistatic to Msn2 in controlling CMK2 expression and the calcium sensitivity of yeast cells in response to calcium stress. This work provides important clues to the study on the regulation of CaMKII expression in mammalian cells.

哺乳动物钙/钙调素依赖性蛋白激酶II （CaMKII）是大脑中的一种记忆分子，调节脂肪酸和脂质代谢。作为CaMKII的酵母同源物，cm2是酿酒酵母钙信号的负反馈调节因子。先前的系统研究表明，在钙胁迫以外的各种条件下，42种转录因子（tf）参与控制CMK2的表达，但据报道只有一种转录因子Crz1在钙胁迫下直接调节CMK2的表达。在这里，我们发现了Adr1、Aft2、Cad1、Cst6、Cup2、Dal81、Dal82、Flo8、Gcr2、Haa1、Hfi1、Msn2、Oaf1、Pho4、Ppr1、Rfx1、Rgm1、Rpn4、Sfp1、SIp3、Smp1、Spt10、Stp1、Sum1、Swi4和Tup1等26个tf参与CMK2转录的阳性控制，其中10个是钙胁迫特异性的。相反，其他四个tf， Hir2, Rph1， Sin3和Uga3，负调控CMK2转录，不依赖于钙胁迫。因此，多种TFs直接或间接地控制酵母细胞中CMK2的转录。EMSA和ChIP分析表明，一般应激响应的Msn2通过启动子中的一个STRE位点5‘ C-155CCCT 3’直接控制cm2的表达。我们的遗传研究表明，Crz1在控制酵母细胞对钙胁迫的CMK2表达和钙敏感性方面是上位性的。这项工作为研究CaMKII在哺乳动物细胞中的表达调控提供了重要线索。

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引用次数: 0

A putative PIWIL/piRNA–OTX2 network: An emerging model for epigenetic regulation of cancer stemness in retinoblastoma PIWIL/piRNA-OTX2网络：视网膜母细胞瘤肿瘤干细胞的表观遗传调控新模型

IF 2.6 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et Biophysica Acta-Gene Regulatory Mechanisms

Pub Date : 2025-07-19 DOI: 10.1016/j.bbagrm.2025.195106

Rupa Roy , Subbulakshmi Chidambaram

Recent findings underscore the critical role of PIWIL/piRNA pathways in cancer, extending their known functions beyond reproductive biology. Retinoblastoma (RB), a rare pediatric retinal tumor, is primarily driven by RB1 gene loss but also involves significant epigenetic alterations. Our previous studies revealed that PIWIL4 is significantly upregulated in RB, and its knockdown disrupts the expression of stemness-associated factors, including OTX2, SOX2, and NANOG. In this review, we have proposed that PIWIL/piRNA complexes might recruit epigenetic modifiers to cis-regulatory modules (CRMs) of the OTX2 gene, modulating chromatin accessibility and transcription factor binding. Aberrant PIWIL4 expression may dysregulate OTX2 expression, impacting stemness-maintaining factors and activating oncogenic pathways, including Wnt/β-catenin signaling, mediated by TSPAN12, EphA2, and ZNF proteins. This conceptual framework positions the PIWIL/piRNA-OTX2 axis as a potential regulator of CSC dynamics, linking it to epigenetic modifications, transcription factor interactions, and neuronal differentiation in RB. Targeting this axis could disrupt stemness-associated pathways and oncogenic signaling, offering new therapeutic strategies to mitigate tumor progression and recurrence in RB and other cancers with similar molecular mechanisms.

最近的研究结果强调了PIWIL/piRNA通路在癌症中的关键作用，将其已知功能扩展到生殖生物学之外。视网膜母细胞瘤（RB）是一种罕见的儿童视网膜肿瘤，主要由RB1基因缺失驱动，但也涉及显著的表观遗传改变。我们之前的研究表明，PIWIL4在RB中显著上调，其下调会破坏OTX2、SOX2和NANOG等stemness相关因子的表达。在这篇综述中，我们提出PIWIL/piRNA复合物可能将表观遗传修饰因子募集到OTX2基因的顺式调控模块（CRMs）中，调节染色质可及性和转录因子结合。PIWIL4的异常表达可能会使OTX2的表达失调，影响干细胞维持因子，激活由TSPAN12、EphA2和ZNF蛋白介导的Wnt/β-catenin信号通路。这一概念框架将PIWIL/piRNA-OTX2轴定位为CSC动力学的潜在调节因子，将其与RB的表观遗传修饰、转录因子相互作用和神经元分化联系起来。靶向该轴可能会破坏干细胞相关通路和致癌信号，为减轻RB和其他具有类似分子机制的癌症的肿瘤进展和复发提供新的治疗策略。

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引用次数: 0

Functional characterization of promoter regions in selenoprotein synthesis-relevant genes (sbp2, eefsec and sepsecs) and their selenium-dependent regulation in yellow catfish Pelteobagrus fulvidraco 黄颡鱼硒蛋白合成相关基因（sbp2、eefsec和sepsecs）启动子区功能特征及其硒依赖性调控

IF 2.6 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et Biophysica Acta-Gene Regulatory Mechanisms

Pub Date : 2025-07-04 DOI: 10.1016/j.bbagrm.2025.195105

Kai Zhang , An-Gen Yu , Hua Zheng , Pei-Jia Li , Zhi Luo

The study explored the transcriptional regulation of selenoprotein synthesis-relevant genes, such as selenocysteine insertion sequence element binding protein 2 (sbp2), eukaryotic elongation factor (eefsec) and o-phosphoserine selenocysteine tRNA synthase (sepsecs), and their selenium-mediated regulation in yellow catfish Pelteobagrus fulvidraco, an important fish with ecological and economic importance in several Asian countries. We cloned the sequences of sbp2, eefsec and sepsecs promoters, spanning from −2060 bp to +61 bp, −1910 bp to +53 bp and − 1456 bp to +51 bp relative to the TSS, respectively. Through sequential deletion and mutation analysis of their promoters, we identified several functional binding sites: the signal transducer and activator of transcription 1 (STAT1) binding site (−1308 bp to −1322 bp) and the forkhead box protein O1 (FOXO1) binding site (−1778 bp to −1788 bp) in the sbp2 promoter; the FOXO1 binding site (−1070 bp to −1080 bp) and the STAT3 binding site (−428 bp to −436 bp) in the eefsec promoter; and the FOXO1 binding site (−721 bp to −731 bp) in the sepsecs promoter. The activity of these binding sites was regulated by selenomethionine (Se-Met) incubation. Furthermore, electrophoretic mobility shift assay and chromatin immunoprecipitation experiments confirmed that these binding sites interact with their corresponding transcription factors above. For the first time, we demonstrated that STAT1 and FOXO1 regulate transcriptional activity of sbp2 promoter; STAT3 and FOXO1 regulate transcriptional activity of eefsec promoter; and FOXO1 regulates transcriptional activity of sepsecs promoter. These findings provide novel insights into regulatory mechanisms of selenoprotein synthesis in yellow catfish.

本研究探讨了硒蛋白合成相关基因，如硒氨酸半胱氨酸插入序列元件结合蛋白2 （sbp2）、真核延伸因子（eefsec）和磷酸丝氨酸硒氨酸半胱氨酸tRNA合酶（sepsecs）在黄颡鱼（Pelteobagrus fulvidraco）中的转录调控及其硒介导的调控。黄颡鱼是亚洲一些国家重要的生态和经济鱼类。我们克隆了sbp2、eefsec和sepsecs启动子的序列，相对于TSS分别为-2060 bp至+61 bp、-1910 bp至+53 bp和 - 1456 bp至+51 bp。通过对其启动子的序列删除和突变分析，我们确定了几个功能结合位点：sbp2启动子中的转录信号转换器和激活子1 （STAT1）结合位点（-1308 bp至-1322 bp）和叉头盒蛋白O1 （FOXO1）结合位点（-1778 bp至-1788 bp）；eefsec启动子中的FOXO1结合位点（-1070 bp ~ -1080 bp）和STAT3结合位点（-428 bp ~ -436 bp）；sepsecs启动子中的FOXO1结合位点（-721 bp至-731 bp）。硒代蛋氨酸（Se-Met）孵育可调节这些结合位点的活性。此外，电泳迁移率转移实验和染色质免疫沉淀实验证实了这些结合位点与上述相应的转录因子相互作用。我们首次证实STAT1和FOXO1调控sbp2启动子的转录活性；STAT3和fox01调控eefsec启动子的转录活性；fox01调控sepsecs启动子的转录活性。这些发现为黄鲶鱼硒蛋白合成的调控机制提供了新的见解。

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引用次数: 0

Transposon insertion causes ctnnb2 transcript instability that results in the maternal effect zebrafish ichabod (ich) mutation 转座子插入导致ctnnb2转录不稳定，导致母体效应斑马鱼ichabod （rich）突变。

IF 2.6 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et Biophysica Acta-Gene Regulatory Mechanisms

Pub Date : 2025-06-30 DOI: 10.1016/j.bbagrm.2025.195104

Zsombor Varga , Ferenc Kagan , Shingo Maegawa , Ágnes Nagy , Javan Okendo , Shawn M. Burgess , Eric S. Weinberg , Máté Varga

The maternal-effect mutation ichabod (ich) results in ventralized zebrafish embryos due to impaired induction of the dorsal canonical Wnt-signaling pathway. While previous studies linked the phenotype to reduced ctnnb2 transcript levels, the causative mutation remained unidentified. Using long-read sequencing, we discovered that the ich phenotype stems from the insertion of a non-autonomous CMC-Enhancer/Suppressor-mutator (CMC-EnSpm) transposon in the 3’UTR of the gene. Through reporter assays, we demonstrate that while wild type ctnnb2 mRNAs exhibit remarkably high stability throughout the early stages of development, the insertion of the transposon dramatically reduces transcript stability. Genome-wide mapping of the CMC-EnSpm transposons across multiple zebrafish strains also indicated ongoing transposition activity in the zebrafish genome. Our findings not only resolve the molecular basis of the ich mutation but also highlight the continuing mutagenic potential of endogenous transposons and reveal unexpected aspects of maternal transcript regulation during early zebrafish development.

母体效应突变ichabod （ich）导致斑马鱼胚胎腹化，这是由于背部典型wnt信号通路的诱导受损。虽然先前的研究将表型与ctnnb2转录物水平降低联系起来，但致病突变仍未确定。通过长读测序，我们发现丰富的表型源于在基因的3'UTR中插入一个非自主的cmc -增强子/抑制子-突变子（CMC-EnSpm）转座子。通过报告者实验，我们发现野生型ctnnb2 mrna在早期发育阶段表现出非常高的稳定性，但转座子的插入显著降低了转录物的稳定性。CMC-EnSpm转座子在多个斑马鱼菌株中的全基因组定位也表明斑马鱼基因组中正在进行转座子活性。我们的发现不仅解决了丰富突变的分子基础，还强调了内源性转座子的持续致突变潜力，并揭示了斑马鱼早期发育过程中母体转录调控的意想不到的方面。

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引用次数: 0

DUSP1 protein's impact on breast cancer: Anticancer response and sensitivity to cisplatin DUSP1蛋白对乳腺癌的影响：抗癌反应和对顺铂的敏感性

IF 2.6 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et Biophysica Acta-Gene Regulatory Mechanisms

Pub Date : 2025-06-25 DOI: 10.1016/j.bbagrm.2025.195103

Sefa Metin , Hilal Altan , Ergün Tercan , Bala Gur Dedeoglu , Hakan Gurdal

Dual-Specificity Phosphatase 1 (DUSP1) modulates the activity of members of the Mitogen-Activated Protein Kinase (MAPK) family, including p38, JNK, and ERK1/2, which affects various cellular functions in cancer. Moreover, DUSP1 is known to influence the outcomes of cancer chemotherapy. This study aimed to reduce DUSP1 protein expression using CRISPR/Cas9 and siRNA and assess its effects on cell proliferation, migration, and tumor growth potential in triple-negative breast cancer (TNBC) cells. We examined the expression levels of p38, JNK, and ERK1/2, along with their phosphorylated forms, and investigated DUSP1's influence to cisplatin sensitivity. Our findings revealed that the downregulation of DUSP1 expression inhibited the proliferation, migration, and tumor growth potential of TNBC cells. Additionally, BCI, an inhibitor of DUSP1/6, demonstrated anti-proliferative effects on these cells. Decreasing the expression of DUSP1 increased the phosphorylation ratio of p38 and JNK, but not ERK1/2. Moreover, the anticancer response induced by cisplatin was enhanced by reducing DUSP1 expression or by treating the cells with BCI. Notably, cisplatin treatment increased p38 phosphorylation, which was significantly augmented by reduced DUSP1 expression. We also demonstrated that the DUSP1 inhibition-induced anticancer response in these cells predominantly relied on p38 activity. These findings contribute to a better understanding of the role of DUSP1 in breast cancer and offer insights into potential therapeutic strategies targeting DUSP1 to enhance the efficacy of cisplatin treatment. Our study highlights that decreased DUSP1 protein expression and activity mediates an anticancer response and increases the sensitivity of MDA-MB231 cells to cisplatin by regulating p38.

双特异性磷酸酶1 （DUSP1）调节丝裂原活化蛋白激酶（MAPK）家族成员的活性，包括p38、JNK和ERK1/2，影响癌症中的各种细胞功能。此外，已知DUSP1会影响癌症化疗的结果。本研究旨在利用CRISPR/Cas9和siRNA降低DUSP1蛋白表达，并评估其对三阴性乳腺癌（TNBC）细胞增殖、迁移和肿瘤生长潜力的影响。我们检测了p38、JNK和ERK1/2及其磷酸化形式的表达水平，并研究了DUSP1对顺铂敏感性的影响。我们的研究结果表明，下调DUSP1的表达抑制了TNBC细胞的增殖、迁移和肿瘤生长潜能。此外，DUSP1/6的抑制剂BCI对这些细胞具有抗增殖作用。降低DUSP1的表达增加了p38和JNK的磷酸化比例，但没有增加ERK1/2的磷酸化比例。此外，顺铂诱导的抗癌反应可通过降低DUSP1表达或BCI处理细胞而增强。值得注意的是，顺铂治疗增加了p38磷酸化，而DUSP1表达的降低则显著增强了p38磷酸化。我们还证明了DUSP1抑制诱导的这些细胞的抗癌反应主要依赖于p38活性。这些发现有助于更好地了解DUSP1在乳腺癌中的作用，并为针对DUSP1的潜在治疗策略提供见解，以提高顺铂治疗的疗效。我们的研究强调，DUSP1蛋白表达和活性的降低介导抗癌反应，并通过调节p38增加MDA-MB231细胞对顺铂的敏感性。

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引用次数: 0

RhoA functionally collaborates with HSPA1A to promote the migratory phenotype of cancer cells RhoA与HSPA1A在功能上协同促进癌细胞的迁移表型

IF 2.6 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et Biophysica Acta-Gene Regulatory Mechanisms

Pub Date : 2025-06-20 DOI: 10.1016/j.bbagrm.2025.195101

Sumaiya Nabi , Mohammad Amin Hajam , Umar Mushtaq , Aadil Manzoor Baba , Bashir Ahmad Malla , Firdous Ahmad Khanday , Nazir Ahmad Dar

RhoA, a member of the GTPase family, plays a pivotal role in attaining a migratory phenotype, mainly by regulating cytoskeleton dynamics, cell adhesion and membrane protrusions. Although many upstream regulators and downstream effectors of RhoA have been identified, the discovery of new interacting partners continues to expand its interactome, providing fresh insights into its regulation and function. Co-immunoprecipitation and fluorescence microscopy were used to study the interaction, localization and morphological effects of HSPA1A and RhoA. The interaction was validated by modulating the protein expression through transfections and silencing approaches. Cell proliferation, migration and viability were assessed using MTT, a Boyden chamber and FACS assays, respectively. Our study identified HSPA1A, as an unexplored interacting partner of RhoA under physiological conditions. Functional analyses showed that the interaction between HSPA1A and RhoA enhances the migratory potential of cancer cells, induces G0/G1 cell cycle arrest and promotes a rounded cell morphology. Under HSPA1A transfection, increased RhoA protein levels were observed, while the silencing of HSPA1A resulted in decreased RhoA levels. This study highlights the critical role of HSPA1A-RhoA interaction in regulating cancer cell migration, morphology and cell cycle progression. These findings lay the groundwork for future research into its potential clinical applications.

RhoA是GTPase家族的一员，主要通过调节细胞骨架动力学、细胞粘附和膜突出在迁移表型的实现中起关键作用。虽然RhoA的许多上游调控因子和下游效应因子已经被确定，但新的相互作用伙伴的发现继续扩大其相互作用组，为其调节和功能提供了新的见解。采用免疫共沉淀法和荧光显微镜技术研究HSPA1A和RhoA的相互作用、定位和形态效应。通过转染和沉默方法调节蛋白表达，验证了这种相互作用。分别采用MTT、Boyden室和FACS法评估细胞增殖、迁移和活力。我们的研究发现HSPA1A在生理条件下是RhoA的一个未被探索的相互作用伙伴。功能分析表明，HSPA1A与RhoA的相互作用增强了癌细胞的迁移潜能，诱导G0/G1细胞周期阻滞，促进细胞形态呈圆形。转染HSPA1A后，RhoA蛋白水平升高，而沉默HSPA1A导致RhoA蛋白水平降低。本研究强调了HSPA1A-RhoA相互作用在调节癌细胞迁移、形态和细胞周期进程中的关键作用。这些发现为进一步研究其潜在的临床应用奠定了基础。

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引用次数: 0

ARID1A-driven modulation of EZH2 impedes proliferation and enhances senescence in breast cancer cells arid1a驱动的EZH2调控可抑制乳腺癌细胞的增殖并促进衰老。

IF 2.6 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et Biophysica Acta-Gene Regulatory Mechanisms

Pub Date : 2025-06-19 DOI: 10.1016/j.bbagrm.2025.195102

Neena George , Rayzel Fernandes , Kapaettu Satyamoorthy

ARID1A mutations, in association with EZH2 overexpression, are linked to various malignancies, particularly those driven by epigenetic dysregulation and associated with therapy resistance. The prevalence of ARID1A mutations is high in ER+ breast cancer, and studies have mainly explored the synthetic lethal effects of these proteins. However, the tumor-suppressive mechanisms of ARID1A are complex and not yet fully understood. In this study, we explored the potential tumor-specific epigenetic antagonism between ARID1A and EZH2 in breast cancer cells, particularly focusing on the modulation of EZH2 by ARID1A through senescence pathway activation. Treatment with DNA-damaging agents induced senescence, which was associated with upregulation of ARID1A expression and a concurrent reduction in EZH2 levels, suggesting a potential role for ARID1A in the induction and maintenance of the senescence phenotype. Overexpression of ARID1A led to reduced EZH2 levels, suppressed cell proliferation in MCF-7 and MDA-MB231 cells, and induced a senescence-like phenotype. These cells exhibited changes in cell-to-cell adhesion, increased filopodium formation, and G0/G1 cell cycle arrest. This antiproliferative effect of ARID1A is mediated through the activation of the p53-p21/p16 axis. Furthermore, ARID1A knockdown-associated downregulation of EZH2 highlights the integral role of ARID1A in destabilizing the expression of EZH2 and contributing to cell cycle arrest. Importantly, we found that dasatinib treatment selectively targeted tumor cells overexpressing ARID1A. These findings provide preliminary insight into the molecular mechanisms by which ARID1A regulates EZH2 and establishes a senescence phenotype, offering valuable directions for developing more effective and personalized treatments.

与EZH2过表达相关的ARID1A突变与各种恶性肿瘤有关，特别是那些由表观遗传失调驱动并与治疗耐药性相关的恶性肿瘤。ARID1A突变在ER+乳腺癌中的患病率很高，研究主要探讨这些蛋白的合成致死作用。然而，ARID1A的肿瘤抑制机制是复杂的，尚未完全了解。在这项研究中，我们探索了ARID1A和EZH2在乳腺癌细胞中潜在的肿瘤特异性表观遗传拮抗作用，特别关注了ARID1A通过衰老途径激活对EZH2的调节。dna损伤剂处理诱导衰老，这与ARID1A表达上调和EZH2水平降低相关，提示ARID1A在诱导和维持衰老表型中可能起作用。ARID1A的过表达导致EZH2水平降低，抑制MCF-7和MDA-MB231细胞的细胞增殖，并诱导衰老样表型。这些细胞表现出细胞间粘附的变化，丝足形成增加，G0/G1细胞周期停滞。ARID1A的这种抗增殖作用是通过激活p53-p21/p16轴介导的。此外，ARID1A敲低相关的EZH2下调强调了ARID1A在破坏EZH2表达稳定和促进细胞周期阻滞中的整体作用。重要的是，我们发现达沙替尼治疗选择性靶向过表达ARID1A的肿瘤细胞。这些发现初步揭示了ARID1A调控EZH2并建立衰老表型的分子机制，为开发更有效和个性化的治疗方法提供了有价值的指导。

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引用次数: 0