Experimental cell research最新文献_第10页

LINC02987 suppression hepatocellular carcinoma progression by modulating autophagy via the miR-338-3p/ATG12 axis 通过miR-338-3p/ATG12轴调节自噬抑制肝细胞癌进展。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114398

Haiyan Fu , Qiuhong Wang , Haiwen Li , Hongjuan Li , Jie Li , Yu Liu , Futao Dang , Lifeng Wang , Xuan Zhang , Yongrui Yang , Yingrong Du

Hepatocellular carcinoma (HCC), the most common primary liver cancer, is marked by a high mortality rate, with the misregulation of long non-coding RNAs (LncRNAs) playing a key role in its development. Here, we studied the role of LINC02987 in HCC. We employed bioinformatics tools to identify LncRNAs and miRNAs that exhibit differential expression in HCC. Quantitative real-time reverse transcription PCR (RT-qPCR) and Western blot analysis were utilized to quantify gene and protein expression levels. The interaction between miR-338-3p and LINC02987 or ATG12 was confirmed through dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. We observed that LINC02987 was overexpressed in HCC tumor tissues and cell lines. Silencing of LINC02987 led to a reduction in cell viability, diminished clonogenic potential, and attenuated invasive and migratory capabilities. Also, decreasing protein level and fluorescence intensity of the autophagy-associated LC3 I/II. In HCC, miR-338-3p expression was downregulated, while inversely correlates with the overexpression of the autophagy protein ATG12. Mimicking miR-338-3p suppresses the activity of both LINC02987 and ATG12, as evidenced by reduced luciferase signals in corresponding reporter assays. Mimicking miR-338-3p suppresses the activity of both LINC02987 and ATG12, as evidenced by reduced luciferase signals in reporter assays. Transfection with si-LINC02987 decreased ATG12 expression, an effect that was partially reversed by miR-338-3p knockdown. Inhibition of miR-338-3p or overexpression of ATG12 increased LC3 I/II protein levels. Our results indicate that LINC02987 sequesters miR-338-3p, leading to increased ATG12 and promoting autophagy in HCC cells. These results highlight the potential of LINC02987 as a therapeutic target for the treatment of HCC.

肝细胞癌（hepatellular carcinoma， HCC）是最常见的原发性肝癌，其特点是死亡率高，长链非编码rna （long non-coding rna, LncRNAs）的错误调控在其发生发展中起着关键作用。在此，我们研究了LINC02987在HCC中的作用。我们使用生物信息学工具来鉴定在HCC中表现出差异表达的lncrna和mirna。采用实时荧光定量反转录PCR （RT-qPCR）和Western blot分析定量基因和蛋白的表达水平。通过双荧光素酶报告基因和RNA免疫沉淀（RIP）实验证实了miR-338-3p与LINC02987或ATG12之间的相互作用。我们观察到LINC02987在HCC肿瘤组织和细胞系中过表达。沉默LINC02987导致细胞活力降低，克隆潜能减弱，侵袭和迁移能力减弱。同时，自噬相关LC3 I/II蛋白水平和荧光强度降低。在HCC中，miR-338-3p表达下调，而与自噬蛋白ATG12的过表达呈负相关。模拟miR-338-3p抑制LINC02987和ATG12的活性，在相应的报告基因检测中，荧光素酶信号降低证明了这一点。模仿miR-338-3p抑制LINC02987和ATG12的活性，报告基因实验中荧光素酶信号的减少证明了这一点。转染si-LINC02987降低ATG12的表达，这一作用被miR-338-3p敲低部分逆转。抑制miR-338-3p或过表达ATG12可增加LC3 I/II蛋白水平。我们的研究结果表明，LINC02987隔离miR-338-3p，导致HCC细胞中ATG12升高并促进自噬。这些结果突出了LINC02987作为HCC治疗靶点的潜力。

{"title":"LINC02987 suppression hepatocellular carcinoma progression by modulating autophagy via the miR-338-3p/ATG12 axis","authors":"Haiyan Fu , Qiuhong Wang , Haiwen Li , Hongjuan Li , Jie Li , Yu Liu , Futao Dang , Lifeng Wang , Xuan Zhang , Yongrui Yang , Yingrong Du","doi":"10.1016/j.yexcr.2024.114398","DOIUrl":"10.1016/j.yexcr.2024.114398","url":null,"abstract":"<div><div>Hepatocellular carcinoma (HCC), the most common primary liver cancer, is marked by a high mortality rate, with the misregulation of long non-coding RNAs (LncRNAs) playing a key role in its development. Here, we studied the role of LINC02987 in HCC. We employed bioinformatics tools to identify LncRNAs and miRNAs that exhibit differential expression in HCC. Quantitative real-time reverse transcription PCR (RT-qPCR) and Western blot analysis were utilized to quantify gene and protein expression levels. The interaction between miR-338-3p and LINC02987 or ATG12 was confirmed through dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. We observed that LINC02987 was overexpressed in HCC tumor tissues and cell lines. Silencing of LINC02987 led to a reduction in cell viability, diminished clonogenic potential, and attenuated invasive and migratory capabilities. Also, decreasing protein level and fluorescence intensity of the autophagy-associated LC3 I/II. In HCC, miR-338-3p expression was downregulated, while inversely correlates with the overexpression of the autophagy protein ATG12. Mimicking miR-338-3p suppresses the activity of both LINC02987 and ATG12, as evidenced by reduced luciferase signals in corresponding reporter assays. Mimicking miR-338-3p suppresses the activity of both LINC02987 and ATG12, as evidenced by reduced luciferase signals in reporter assays. Transfection with si-LINC02987 decreased ATG12 expression, an effect that was partially reversed by miR-338-3p knockdown. Inhibition of miR-338-3p or overexpression of ATG12 increased LC3 I/II protein levels. Our results indicate that LINC02987 sequesters miR-338-3p, leading to increased ATG12 and promoting autophagy in HCC cells. These results highlight the potential of LINC02987 as a therapeutic target for the treatment of HCC.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"444 2","pages":"Article 114398"},"PeriodicalIF":3.3,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142921462","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

RPF2 and CARM1 cooperate to enhance colorectal cancer metastasis via the AKT/GSK-3β signaling pathway RPF2和CARM1通过AKT/GSK-3β信号通路共同促进结直肠癌转移。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114374

Cong Cheng , KeMing Zhang , MaCheng Lu , Yuan Zhang , Tong Wang , Ye Zhang

RPF2 plays a crucial role in promoting epithelial-mesenchymal transition (EMT) and regulating metastasis in colorectal cancer (CRC). By analyzing data from the TCGA and GEO databases, we observed significantly elevated RPF2 expression in CRC, which correlated with EMT markers. Further investigations using stable RPF2 overexpression and knockdown cell lines demonstrated that RPF2 facilitates EMT activation through the AKT/GSK-3β signaling pathway. Notably, CARM1 was identified as a key downstream effector of RPF2. Selective inhibition of CARM1 effectively suppressed the activation of the AKT/GSK-3β pathway and EMT induced by RPF2 overexpression. Both in vitro and in vivo experiments confirmed that RPF2 expression levels positively correlate with the metastatic potential of CRC cells. Moreover, treatment with a CARM1 inhibitor significantly reduced the invasive and migratory capabilities of RPF2-overexpressing cells. These findings suggest that RPF2 drives CRC metastasis by modulating EMT via the AKT/GSK-3β pathway, with CARM1 serving as a critical mediator, offering potential therapeutic targets for CRC.

RPF2在促进结直肠癌（CRC）上皮-间质转化（EMT）和调节转移中起着至关重要的作用。通过分析TCGA和GEO数据库的数据，我们观察到RPF2在CRC中的表达显著升高，其与EMT标志物相关。对稳定的RPF2过表达和敲低细胞系的进一步研究表明，RPF2通过AKT/GSK-3β信号通路促进EMT激活。值得注意的是，CARM1被确定为RPF2的关键下游效应因子。选择性抑制CARM1可有效抑制AKT/GSK-3β通路的激活和RPF2过表达诱导的EMT。体外和体内实验均证实RPF2表达水平与CRC细胞的转移潜能呈正相关。此外，CARM1抑制剂显著降低了rpf2过表达细胞的侵袭和迁移能力。这些发现表明，RPF2通过AKT/GSK-3β通路调节EMT驱动结直肠癌转移，而CARM1作为关键介质，为结直肠癌提供了潜在的治疗靶点。

{"title":"RPF2 and CARM1 cooperate to enhance colorectal cancer metastasis via the AKT/GSK-3β signaling pathway","authors":"Cong Cheng , KeMing Zhang , MaCheng Lu , Yuan Zhang , Tong Wang , Ye Zhang","doi":"10.1016/j.yexcr.2024.114374","DOIUrl":"10.1016/j.yexcr.2024.114374","url":null,"abstract":"<div><div>RPF2 plays a crucial role in promoting epithelial-mesenchymal transition (EMT) and regulating metastasis in colorectal cancer (CRC). By analyzing data from the TCGA and GEO databases, we observed significantly elevated RPF2 expression in CRC, which correlated with EMT markers. Further investigations using stable RPF2 overexpression and knockdown cell lines demonstrated that RPF2 facilitates EMT activation through the AKT/GSK-3β signaling pathway. Notably, CARM1 was identified as a key downstream effector of RPF2. Selective inhibition of CARM1 effectively suppressed the activation of the AKT/GSK-3β pathway and EMT induced by RPF2 overexpression. Both <em>in vitro</em> and <em>in vivo</em> experiments confirmed that RPF2 expression levels positively correlate with the metastatic potential of CRC cells. Moreover, treatment with a CARM1 inhibitor significantly reduced the invasive and migratory capabilities of RPF2-overexpressing cells. These findings suggest that RPF2 drives CRC metastasis by modulating EMT via the AKT/GSK-3β pathway, with CARM1 serving as a critical mediator, offering potential therapeutic targets for CRC.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"444 2","pages":"Article 114374"},"PeriodicalIF":3.3,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142824070","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Nucleostemin interacts with SMAD3 promoting tumor metastasis 核干素与SMAD3相互作用促进肿瘤转移。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114362

Xuling Sun , Jiageng He , Yujiang Li , Zhiqiang Chu , Lei Zhu , Hui Zhang , Xiangwei Wu

SMAD3 plays a crucial role in TGF-β, regulating various normal developmental mechanisms and disease pathogenesis. Here, we report that SMAD3 directly interacts with Nucleostemin (NS), leading to nuclear translocation and affecting SMAD3 activity after TGF-β1 stimulation. Moreover, NS acts as a competitor, preventing PPM1A from recognizing and dephosphorylating SMAD3. Experimental investigations have demonstrated that NS significantly enhances cellular migration and invasion by promoting the EMT mechanism in vitro. NS knockdown notably suppresses tumor metastasis in the lungs and liver in vivo. Importantly, NS expression is significantly elevated in numerous human malignancies, correlating with a poorer prognosis. The collective evidence from these studies suggests that NS exhibits oncogenic characteristics, supporting further exploration of NS as a potential target for tumor treatment.

SMAD3在TGF-β中发挥重要作用，调节多种正常发育机制和疾病发病机制。在这里，我们报道SMAD3直接与核干素（NS）相互作用，导致核易位，并在TGF-β1刺激后影响SMAD3的活性。此外，NS作为竞争者，阻止PPM1A识别和去磷酸化SMAD3。实验研究表明，NS通过促进体外EMT机制显著增强细胞迁移和侵袭。NS敲低在体内可显著抑制肿瘤在肺和肝脏的转移。重要的是，NS表达在许多人类恶性肿瘤中显著升高，与较差的预后相关。这些研究的共同证据表明，NS具有致癌特征，支持进一步探索NS作为肿瘤治疗的潜在靶点。

引用次数: 0

GPC-3 in hepatocellular carcinoma; A novel biomarker and molecular target GPC-3在肝细胞癌中的表达；一种新的生物标志物和分子靶点。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114391

Hamed Azhdari Tehrani , Masood Zangi , Mobina Fathi , Kimia Vakili , Moustapha Hassan , Elham Rismani , Nikoo Hossein-Khannazer , Massoud Vosough

Hepatocellular carcinoma (HCC) is a global health issue due to its late diagnosis and high recurrence rate. The early detection and diagnosis of HCC with specific and sensitive biomarkers and using novel treatment approaches to improve patient outcomes are essential. Glypican-3 (GPC-3) is a cell surface proteoglycan that is overexpressed in many tumors, including HCC. GPC-3 could be used as a specific biomarker for HCC early detection and could be a potential target for precise therapeutic strategies. Effective identification of GPC-3 could improve both diagnosis and targeted therapy of HCC. Moreover, targeted therapy using GPC-3 could result in a better treatment outcome. Recently, GPC3-targeted therapies have been used in different investigational therapeutic approaches like bi-specific/monoclonal antibodies, peptide vaccines, and CAR T cell therapies. This study aims to highlight the theranostic potential of GPC-3 as a novel biomarker for early detection and as a potential molecular target for HCC treatment as well.

肝细胞癌（HCC）因其诊断晚、复发率高而成为一个全球性的健康问题。HCC的早期检测和诊断具有特异性和敏感性的生物标志物，并使用新的治疗方法来改善患者的预后是至关重要的。Glypican-3 （GPC-3）是一种细胞表面蛋白聚糖，在包括HCC在内的许多肿瘤中过表达。GPC-3可以作为HCC早期检测的特异性生物标志物，并可能成为精确治疗策略的潜在靶点。GPC-3的有效鉴定有助于肝癌的诊断和靶向治疗。此外，使用GPC-3进行靶向治疗可能会产生更好的治疗效果。最近，gpc3靶向治疗已被用于不同的研究性治疗方法，如双特异性/单克隆抗体、肽疫苗和CAR - T细胞治疗。本研究旨在强调GPC-3作为早期检测的新型生物标志物和HCC治疗的潜在分子靶点的治疗潜力。

{"title":"GPC-3 in hepatocellular carcinoma; A novel biomarker and molecular target","authors":"Hamed Azhdari Tehrani , Masood Zangi , Mobina Fathi , Kimia Vakili , Moustapha Hassan , Elham Rismani , Nikoo Hossein-Khannazer , Massoud Vosough","doi":"10.1016/j.yexcr.2024.114391","DOIUrl":"10.1016/j.yexcr.2024.114391","url":null,"abstract":"<div><div>Hepatocellular carcinoma (HCC) is a global health issue due to its late diagnosis and high recurrence rate. The early detection and diagnosis of HCC with specific and sensitive biomarkers and using novel treatment approaches to improve patient outcomes are essential. Glypican-3 (GPC-3) is a cell surface proteoglycan that is overexpressed in many tumors, including HCC. GPC-3 could be used as a specific biomarker for HCC early detection and could be a potential target for precise therapeutic strategies. Effective identification of GPC-3 could improve both diagnosis and targeted therapy of HCC. Moreover, targeted therapy using GPC-3 could result in a better treatment outcome. Recently, GPC3-targeted therapies have been used in different investigational therapeutic approaches like bi-specific/monoclonal antibodies, peptide vaccines, and CAR T cell therapies. This study aims to highlight the theranostic potential of GPC-3 as a novel biomarker for early detection and as a potential molecular target for HCC treatment as well.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"444 2","pages":"Article 114391"},"PeriodicalIF":3.3,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142893398","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Müller glia in short-term dark adaptation of the Austrolebias charrua retina: Cell proliferation and cytoarchitecture 视网膜短期暗适应中的突触神经胶质：细胞增殖和细胞结构。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114394

Laura Herrera-Astorga , Stephanie Silva , Inés Berrosteguieta , Juan Carlos Rosillo , Anabel Sonia Fernández

Fish with unique life cycles offer valuable insights into retinal plasticity, revealing mechanisms of environmental adaptation, cell proliferation, and thus, potentially regeneration. The variability of the environmental factors to which Austrolebias annual fishes are exposed has acted as a strong selective pressure shaping traits such as nervous system plasticity. This has contributed to adaptation to their extreme conditions including the decreased luminosity as ponds dry out. In particular, the retina of A. charrua has been shown to respond to 30 days of decreased luminosity by exacerbating cell proliferation Now, we aimed to determine the cellular component of the retina involved in shorter-term responses. To this end, we performed 5-bromo-2′-deoxyuridine (BrdU) experiments, exposing adult fish to a short period (11 days) of constant darkness. Strikingly, in control conditions, neurogenesis in the inner nuclear and ganglion cell layer in the differentiated retina was detected. In constant darkness, we observed an effect on inner nuclear layer cell proliferation and changes in retinal cytoarchitecture of the retina with cell clusters located in the inner plexiform layer. Additionally, increased BLBP (brain lipid-binding protein) presence was detected in darkness, which has been previously associated with immature and reactivated Müller glia. Thus, our results suggest that the A. charrua retina can respond to environmental changes via rapid activation of progenitor cells in the INL, namely the Müller glia This leads us to hypothesize, that cell proliferation and neurogenesis might contribute to the responses to the functional needs of organisms, potentially playing an adaptive role.

具有独特生命周期的鱼类为研究视网膜可塑性提供了有价值的见解，揭示了环境适应、细胞增殖以及潜在再生的机制。黄鳝所处环境因子的变异性是形成其神经系统可塑性等特征的一种强大的选择压力。这有助于适应它们的极端条件，包括池塘干涸时亮度下降。特别的是，研究表明，在30天的亮度下降后，黄花蒿的视网膜会通过加剧细胞增殖来做出反应。现在，我们的目标是确定视网膜中参与短期反应的细胞成分。为此，我们进行了5-溴-2'-脱氧尿嘧啶（BrdU）实验，将成年鱼暴露在短时间（11天）的持续黑暗中。引人注目的是，在对照条件下，在分化视网膜的细胞核和神经节细胞层中检测到神经发生。我们观察到持续黑暗对视网膜内核层细胞增殖的影响和视网膜细胞结构的变化，细胞团位于视网膜内丛状层。此外，在黑暗中检测到BLBP（脑脂结合蛋白）的增加，这与未成熟和重新激活的神经胶质细胞有关。因此，我们的研究结果表明，A. charrua视网膜可以通过快速激活INL中的祖细胞（即m ller胶质细胞）来响应环境变化。这使我们假设，细胞增殖和神经发生可能有助于对生物体功能需求的反应，潜在地发挥适应性作用。

{"title":"Müller glia in short-term dark adaptation of the Austrolebias charrua retina: Cell proliferation and cytoarchitecture","authors":"Laura Herrera-Astorga , Stephanie Silva , Inés Berrosteguieta , Juan Carlos Rosillo , Anabel Sonia Fernández","doi":"10.1016/j.yexcr.2024.114394","DOIUrl":"10.1016/j.yexcr.2024.114394","url":null,"abstract":"<div><div>Fish with unique life cycles offer valuable insights into retinal plasticity, revealing mechanisms of environmental adaptation, cell proliferation, and thus, potentially regeneration. The variability of the environmental factors to which Austrolebias annual fishes are exposed has acted as a strong selective pressure shaping traits such as nervous system plasticity. This has contributed to adaptation to their extreme conditions including the decreased luminosity as ponds dry out. In particular, the retina of <em>A. charrua</em> has been shown to respond to 30 days of decreased luminosity by exacerbating cell proliferation Now, we aimed to determine the cellular component of the retina involved in shorter-term responses. To this end, we performed 5-bromo-2′-deoxyuridine (BrdU) experiments, exposing adult fish to a short period (11 days) of constant darkness. Strikingly, in control conditions, neurogenesis in the inner nuclear and ganglion cell layer in the differentiated retina was detected. In constant darkness, we observed an effect on inner nuclear layer cell proliferation and changes in retinal cytoarchitecture of the retina with cell clusters located in the inner plexiform layer. Additionally, increased BLBP (brain lipid-binding protein) presence was detected in darkness, which has been previously associated with immature and reactivated Müller glia. Thus, our results suggest that the <em>A. charrua</em> retina can respond to environmental changes via rapid activation of progenitor cells in the INL, namely the Müller glia This leads us to hypothesize, that cell proliferation and neurogenesis might contribute to the responses to the functional needs of organisms, potentially playing an adaptive role.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"444 2","pages":"Article 114394"},"PeriodicalIF":3.3,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142893399","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

FoxC1 activates Notch3 signaling to promote the inflammatory phenotype of keloid fibroblasts and aggravates keloid FoxC1激活Notch3信号，促进瘢痕疙瘩成纤维细胞的炎症表型，加重瘢痕疙瘩。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114402

Yin Wang , Zhengguo Xia , Wengting Wang , Jingsong Zhang , Chao Hu , Fan Wang , Fei Zhu , Lin sen Fang , Jun Wang , Xiaojing Li

Keloids are disfiguring proliferative scars, and their pathological mechanisms are still unclear. We have previously established that FoxC1 plays a significant role in rheumatoid arthritis and osteoarthritis, but its molecular mechanisms in pathological scar formation remain elusive. In this study, we analyzed keloid tissue characteristics using HE staining and immunohistochemistry, revealing abnormal expression of FoxC1 and Notch3 in keloids. Lentiviral modulation of FoxC1 and Notch3 demonstrated that they promote the expression of α-SMA, fibronectin, collagen I, and Hes-1, enhancing the proliferation, migration, invasion, and cytokine production of keloid fibroblasts (KFs) while inhibiting apoptosis. Co-immunoprecipitation (CO-IP), dual-luciferase reporter assays, and chromatin immunoprecipitation (ChIP) confirmed that FoxC1 can directly bind to the Notch3 promoter and enhance its transcription. Additionally, in vivo, overexpression of FoxC1 and Notch3 promoted keloid formation. In summary, our research highlights the critical regulatory role of FoxC1 in keloid formation through Notch3 activation, potentially offering new therapeutic targets for preventing scar formation.

瘢痕疙瘩是毁容的增殖性疤痕，其病理机制尚不清楚。我们之前已经确定FoxC1在类风湿关节炎和骨关节炎中起重要作用，但其在病理性瘢痕形成中的分子机制尚不清楚。本研究通过HE染色和免疫组化分析瘢痕疙瘩组织特征，发现FoxC1和Notch3在瘢痕疙瘩组织中表达异常。慢病毒对FoxC1和Notch3的调节表明，它们促进α-SMA、纤维连接蛋白、I型胶原和Hes-1的表达，增强瘢痕疙瘩成纤维细胞（KFs）的增殖、迁移、侵袭和细胞因子的产生，同时抑制细胞凋亡。共免疫沉淀（CO-IP）、双荧光素酶报告基因试验和染色质免疫沉淀（ChIP）证实FoxC1可以直接结合Notch3启动子并增强其转录。此外，在体内，FoxC1和Notch3的过表达促进了瘢痕疙瘩的形成。总之，我们的研究强调了FoxC1通过Notch3激活在瘢痕疙瘩形成中的关键调节作用，可能为预防瘢痕形成提供新的治疗靶点。

{"title":"FoxC1 activates Notch3 signaling to promote the inflammatory phenotype of keloid fibroblasts and aggravates keloid","authors":"Yin Wang , Zhengguo Xia , Wengting Wang , Jingsong Zhang , Chao Hu , Fan Wang , Fei Zhu , Lin sen Fang , Jun Wang , Xiaojing Li","doi":"10.1016/j.yexcr.2024.114402","DOIUrl":"10.1016/j.yexcr.2024.114402","url":null,"abstract":"<div><div>Keloids are disfiguring proliferative scars, and their pathological mechanisms are still unclear. We have previously established that FoxC1 plays a significant role in rheumatoid arthritis and osteoarthritis, but its molecular mechanisms in pathological scar formation remain elusive. In this study, we analyzed keloid tissue characteristics using HE staining and immunohistochemistry, revealing abnormal expression of FoxC1 and Notch3 in keloids. Lentiviral modulation of FoxC1 and Notch3 demonstrated that they promote the expression of α-SMA, fibronectin, collagen I, and Hes-1, enhancing the proliferation, migration, invasion, and cytokine production of keloid fibroblasts (KFs) while inhibiting apoptosis. Co-immunoprecipitation (CO-IP), dual-luciferase reporter assays, and chromatin immunoprecipitation (ChIP) confirmed that FoxC1 can directly bind to the Notch3 promoter and enhance its transcription. Additionally, in vivo, overexpression of FoxC1 and Notch3 promoted keloid formation. In summary, our research highlights the critical regulatory role of FoxC1 in keloid formation through Notch3 activation, potentially offering new therapeutic targets for preventing scar formation.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"444 2","pages":"Article 114402"},"PeriodicalIF":3.3,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142926967","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Mechanism of METTL3 in the proliferation, invasion, and migration of intrahepatic cholangiocarcinoma cells via m6A modification 通过m6A修饰，MEETL3参与肝内胆管癌细胞增殖、侵袭和迁移的机制。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114353

Xinmiao Jiang , Hui Tan

Intrahepatic cholangiocarcinoma (ICC) is a primary invasive malignant tumor. This study was conducted to explore the role of methyltransferase-like 3 (METTL3)-mediated m6A modification in ICC cells and provide novel targets for ICC treatment. Levels of METTL3/YTH N6-methyladenosine RNA binding protein 2 (YTHDF2)/Nedd4 family interacting protein 1 (NDFIP1) in cells were determined. Cell viability, proliferation, invasion, and migration were evaluated. The enrichments of METTL3, YTHDF2, and m6A on NDFIP1 mRNA were analyzed. The mRNA stability was determined. Inhibition of YTHDF2 or NDFIP1 was combined with si-METTL3 to confirm the mechanism. The role of METTL3 in vivo was verified. METTL3 was overexpressed in ICC cells. METTL3 silencing suppressed ICC cell malignant behaviors, which were reversed by METTL3 overexpression. METTL3 increased m6A modification on NDFIP1 mRNA, facilitated YTHDF2 recognition of m6A, and promoted NDFIP1 mRNA degradation, thereby suppressing NDFIP1 expression. YTHDF2 inhibition increased NDFIP1 mRNA levels. NDFIP1 downregulation partially reversed the inhibitory effects of si-METTL3 on ICC cell behaviors, while NDFIP1 overexpression partially reversed the promotive effects of METTL3 on ICC cell behaviors. METTL3 downregulation suppressed ICC growth by increasing NDFIP1 expression. In conclusion, METTL3 aggravates ICC cell proliferation, invasion, and migration by promoting the degradation of NDFIP1 mRNA in a YTHDF2-dependent manner.

肝内胆管癌（ICC）是一种原发性侵袭性恶性肿瘤。本研究旨在探讨甲基转移酶样3 （METTL3）介导的m6A修饰在ICC细胞中的作用，并为ICC治疗提供新的靶点。检测细胞中METTL3/YTH n6 -甲基腺苷RNA结合蛋白2 (YTHDF2)/Nedd4家族相互作用蛋白1 （NDFIP1）水平。评估细胞活力、增殖、侵袭和迁移。分析METTL3、YTHDF2和m6A在NDFIP1 mRNA上的富集程度。测定mRNA的稳定性。将抑制YTHDF2或NDFIP1与si-METTL3联合进行机制验证。验证了METTL3在体内的作用。METTL3在ICC细胞中过表达。METTL3的沉默抑制了ICC细胞的恶性行为，而METTL3的过表达则逆转了这种恶性行为。METTL3增加了m6A对NDFIP1 mRNA的修饰，促进YTHDF2对m6A的识别，促进NDFIP1 mRNA的降解，从而抑制NDFIP1的表达。YTHDF2抑制增加NDFIP1 mRNA水平。NDFIP1下调部分逆转了si-METTL3对ICC细胞行为的抑制作用，NDFIP1过表达部分逆转了METTL3对ICC细胞行为的促进作用。METTL3下调通过增加NDFIP1表达抑制ICC生长。综上所述，METTL3通过以ythdf2依赖的方式促进NDFIP1 mRNA的降解，从而加剧ICC细胞的增殖、侵袭和迁移。

{"title":"Mechanism of METTL3 in the proliferation, invasion, and migration of intrahepatic cholangiocarcinoma cells via m6A modification","authors":"Xinmiao Jiang , Hui Tan","doi":"10.1016/j.yexcr.2024.114353","DOIUrl":"10.1016/j.yexcr.2024.114353","url":null,"abstract":"<div><div>Intrahepatic cholangiocarcinoma (ICC) is a primary invasive malignant tumor. This study was conducted to explore the role of methyltransferase-like 3 (METTL3)-mediated m6A modification in ICC cells and provide novel targets for ICC treatment. Levels of METTL3/YTH N6-methyladenosine RNA binding protein 2 (YTHDF2)/Nedd4 family interacting protein 1 (NDFIP1) in cells were determined. Cell viability, proliferation, invasion, and migration were evaluated. The enrichments of METTL3, YTHDF2, and m6A on NDFIP1 mRNA were analyzed. The mRNA stability was determined. Inhibition of YTHDF2 or NDFIP1 was combined with si-METTL3 to confirm the mechanism. The role of METTL3 <em>in vivo</em> was verified. METTL3 was overexpressed in ICC cells. METTL3 silencing suppressed ICC cell malignant behaviors, which were reversed by METTL3 overexpression. METTL3 increased m6A modification on NDFIP1 mRNA, facilitated YTHDF2 recognition of m6A, and promoted NDFIP1 mRNA degradation, thereby suppressing NDFIP1 expression. YTHDF2 inhibition increased NDFIP1 mRNA levels. NDFIP1 downregulation partially reversed the inhibitory effects of si-METTL3 on ICC cell behaviors, while NDFIP1 overexpression partially reversed the promotive effects of METTL3 on ICC cell behaviors. METTL3 downregulation suppressed ICC growth by increasing NDFIP1 expression. In conclusion, METTL3 aggravates ICC cell proliferation, invasion, and migration by promoting the degradation of NDFIP1 mRNA in a YTHDF2-dependent manner.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"444 2","pages":"Article 114353"},"PeriodicalIF":3.3,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142750419","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The aberrantly activated AURKB supports and complements the function of AURKA in CALR mutated cells through regulating the cell growth and differentiation 异常激活的AURKB通过调节CALR突变细胞的生长和分化来支持和补充AURKA的功能。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114377

Xueting Hu , Xiangru Yu , Liwei Zhang , Qigang Zhang , Mengchu Ji , Kunming Qi , Shujin Wang , Zhenyu Li , Kailin Xu , Chunling Fu

Aurora kinase B (AURKB) was reported to assist Aurora kinase A (AURKA) to regulate cellular mitosis. AURKA has been found activated in myeloproliferative neoplasms (MPNs) patients with CALR gene mutation, however, it's unclear whether AURKB displays a compensatory function of AURKA in regulation of CALR mutant cell growth and differentiation. Here, we found that AURKB, similar with AURKA, was aberrantly activated in CALR mutant patients, and displayed a more tolerance to the aurora kinase inhibitor. Inhibition of AURKA decreased cell growth and colony formation, induced cell differentiation and apoptosis, while, this inhibitive degree was further enhanced when AURKB was blocked by incremental inhibitor. Transcriptomic analyses revealed a more significant gene enrichment in cells with knockdown of AURKB than that of AURKA, mainly reflecting in oxidative phosphorylation, mitosis, proliferation and apoptosis signaling pathway. Moreover, downregulation of AURKB enhanced cell growth arrest and apoptosis more obviously than that of AURKA, and additionally promoted cell differentiation and metabolism-oxygen consumption rate (OCR). Otherwise, overexpression of AURKA or AURKB facilitated the cell proliferation of CALR mutant cells, and made cells more sensitive to the aurora kinase inhibitor. These results suggest that activated AURKB not only supports the functions of AURKA in promoting the growth of CALR mutated cells, but also has impeded the differentiation of these cells.

据报道，极光激酶B （AURKB）协助极光激酶A （AURKA）调节细胞有丝分裂。AURKA在CALR基因突变的骨髓增生性肿瘤（mpn）患者中被激活，但AURKB是否在调节CALR突变细胞的生长和分化中表现出AURKA的代偿功能尚不清楚。在这里，我们发现AURKB与AURKA相似，在CALR突变患者中被异常激活，并且对极光激酶抑制剂表现出更强的耐受性。抑制AURKA抑制了细胞生长和集落形成，诱导细胞分化和凋亡，而当增加抑制剂阻断AURKB时，这种抑制程度进一步增强。转录组学分析显示，AURKB敲低的细胞比AURKA敲低的细胞有更显著的基因富集，主要体现在氧化磷酸化、有丝分裂、增殖和凋亡信号通路。此外，与AURKA相比，AURKB的下调更明显地增强了细胞的生长停滞和凋亡，并促进了细胞的分化和代谢耗氧率（OCR）。否则，AURKA或AURKB的过表达促进了CALR突变细胞的细胞增殖，并使细胞对极光激酶抑制剂更敏感。这些结果表明，激活的AURKB不仅支持AURKA促进CALR突变细胞生长的功能，而且还会阻碍这些细胞的分化。

{"title":"The aberrantly activated AURKB supports and complements the function of AURKA in CALR mutated cells through regulating the cell growth and differentiation","authors":"Xueting Hu , Xiangru Yu , Liwei Zhang , Qigang Zhang , Mengchu Ji , Kunming Qi , Shujin Wang , Zhenyu Li , Kailin Xu , Chunling Fu","doi":"10.1016/j.yexcr.2024.114377","DOIUrl":"10.1016/j.yexcr.2024.114377","url":null,"abstract":"<div><div>Aurora kinase B (AURKB) was reported to assist Aurora kinase A (AURKA) to regulate cellular mitosis. AURKA has been found activated in myeloproliferative neoplasms (MPNs) patients with CALR gene mutation, however, it's unclear whether AURKB displays a compensatory function of AURKA in regulation of CALR mutant cell growth and differentiation. Here, we found that AURKB, similar with AURKA, was aberrantly activated in CALR mutant patients, and displayed a more tolerance to the aurora kinase inhibitor. Inhibition of AURKA decreased cell growth and colony formation, induced cell differentiation and apoptosis, while, this inhibitive degree was further enhanced when AURKB was blocked by incremental inhibitor. Transcriptomic analyses revealed a more significant gene enrichment in cells with knockdown of AURKB than that of AURKA, mainly reflecting in oxidative phosphorylation, mitosis, proliferation and apoptosis signaling pathway. Moreover, downregulation of AURKB enhanced cell growth arrest and apoptosis more obviously than that of AURKA, and additionally promoted cell differentiation and metabolism-oxygen consumption rate (OCR). Otherwise, overexpression of AURKA or AURKB facilitated the cell proliferation of CALR mutant cells, and made cells more sensitive to the aurora kinase inhibitor. These results suggest that activated AURKB not only supports the functions of AURKA in promoting the growth of CALR mutated cells, but also has impeded the differentiation of these cells.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"444 2","pages":"Article 114377"},"PeriodicalIF":3.3,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142871947","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Delactylation diminished the growth inhibitory role of CA3 by restoring DUOX2 expression in hepatocellular carcinoma 去乙酰化通过恢复DUOX2在肝细胞癌中的表达来减弱CA3的生长抑制作用。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114392

Jun Yan , Yunfei Zhou , Jianwen Xu , Yihong Dong , Xun Yang , Xinxin Yang , Aodi Wu , Shuyuan Chang , Yumeng Wang , Qingxin Zhang , Tomii Ayaka , Lei Yu , Liuyang Zhao , Hongxue Meng , Dabin Liu

Lactylation is an emerging pathogenesis of hepatocellular carcinoma (HCC). However, the underlying mechanisms and biological significance remain poorly understood. The Carbonic anhydrase III (CA3) gene, previously defined as a binding protein of SQLE and involved in the NAFLD disease, has now been identified as a novel tumor suppressor in HCC. mRNA expression of CA3 is associated with a favorable prognosis and negatively correlated with serum lactate levels, whereas CA3 protein expression does not correlate with patient prognosis or serum lactate levels, suggested there has lactate-related post-translational modification of CA3 in HCC. Overexpression of CA3 induces cell apoptosis, thereby reducing intracellular reactive oxygen stress (ROS) through the inhibition of DUOX2 expression. The decreased lactylation level of CA3 protein at the K36 residues, induced by SQLE, results in the loss of the anti-cancer effect of CA3. Together, this study has demonstrated that CA3 is a novel tumor suppressor in HCC, and delactylation of CA3 represents a newly identified mechanism by which HCC cells evade growth suppressors.

乳酸化是肝细胞癌（HCC）的一种新发病机制。然而，潜在的机制和生物学意义仍然知之甚少。碳酸酐酶III （CA3）基因，以前被定义为SQLE的结合蛋白并参与NAFLD疾病，现在已被确定为HCC中的一种新的肿瘤抑制因子。CA3 mRNA表达与预后良好相关，且与血清乳酸水平呈负相关，而CA3蛋白表达与患者预后和血清乳酸水平无相关性，提示HCC中存在与乳酸相关的翻译后修饰。CA3过表达诱导细胞凋亡，从而通过抑制DUOX2表达降低细胞内活性氧应激（ROS）。SQLE诱导CA3蛋白在K36残基处的乳酸化水平降低，导致CA3抗癌作用丧失。总之，本研究证明了CA3在HCC中是一种新的肿瘤抑制因子，CA3的去乙酰化代表了HCC细胞逃避生长抑制因子的一种新发现的机制。

{"title":"Delactylation diminished the growth inhibitory role of CA3 by restoring DUOX2 expression in hepatocellular carcinoma","authors":"Jun Yan , Yunfei Zhou , Jianwen Xu , Yihong Dong , Xun Yang , Xinxin Yang , Aodi Wu , Shuyuan Chang , Yumeng Wang , Qingxin Zhang , Tomii Ayaka , Lei Yu , Liuyang Zhao , Hongxue Meng , Dabin Liu","doi":"10.1016/j.yexcr.2024.114392","DOIUrl":"10.1016/j.yexcr.2024.114392","url":null,"abstract":"<div><div>Lactylation is an emerging pathogenesis of hepatocellular carcinoma (HCC). However, the underlying mechanisms and biological significance remain poorly understood. The Carbonic anhydrase III (CA3) gene, previously defined as a binding protein of SQLE and involved in the NAFLD disease, has now been identified as a novel tumor suppressor in HCC. mRNA expression of CA3 is associated with a favorable prognosis and negatively correlated with serum lactate levels, whereas CA3 protein expression does not correlate with patient prognosis or serum lactate levels, suggested there has lactate-related post-translational modification of CA3 in HCC. Overexpression of CA3 induces cell apoptosis, thereby reducing intracellular reactive oxygen stress (ROS) through the inhibition of DUOX2 expression. The decreased lactylation level of CA3 protein at the K36 residues, induced by SQLE, results in the loss of the anti-cancer effect of CA3. Together, this study has demonstrated that CA3 is a novel tumor suppressor in HCC, and delactylation of CA3 represents a newly identified mechanism by which HCC cells evade growth suppressors.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"444 2","pages":"Article 114392"},"PeriodicalIF":3.3,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142876275","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Unraveling the mechanism of microRNA-134 in colon cancer progression: Targeting KRAS and PIK3CA for cell cycle control and histone deacetylase regulation 揭示microRNA-134在结肠癌进展中的机制：靶向KRAS和PIK3CA进行细胞周期控制和组蛋白去乙酰化酶调控

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114385

Ganesan Jothimani , Diptimayee Das , Surajit Pathak , Sarubala Malayaperumal , Hong Zhang , Xiao-Feng Sun , Antara Banerjee

Colon cancer is the leading cause of cancer-related deaths worldwide. MicroRNAs (miRNAs) are key regulators of gene expression, often dysregulated in colon cancer. This study aims to elucidate the therapeutic role of miR-134-5p as a tumor suppressor miRNA in colon cancer cells. We analyzed miRNA expression profiles in primary and metastatic colon cancer cells. The clinical significance of miR-134-5p was evaluated using the TCGA database. Bioinformatics tools (HADDOCK) predicted miRNA-mRNA interactions and the molecular docking of miRNA-mRNA-AGO2 complexes. Luciferase reporter assays, cell proliferation, immunofluorescence, colony forming unit assays, and qRT-PCR analysis assessed miR-134-5p effects on KRAS, PIK3CA, and downstream signaling pathways in primary and metastatic colon cancer cells. miR-134-5p was downregulated in colon cancer cells. Bioinformatics analysis suggested KRAS, PIK3CA, EGFR, and HDAC5 as potential targets. HADDOCK analysis revealed strong binding affinity and structural stability between KRAS, PIK3CA, miR-134-5p, and AGO2. Gene-reporter assays confirmed miR-134-5p-mediated degradation of KRAS and PIK3CA. miR-134-5p transfection reduced KRAS and PI3K protein levels, suppressed EGFR/RTK signaling and its downstream targets, and inhibited HDAC expression, ultimately reducing colon cancer cell proliferation. The results of this study confirm that miR-134-5p acts as a potential tumor suppressor miRNA in colon cancer cells by inhibiting KRAS and PI3K expression through AGO2-mediated gene silencing. It deregulates downstream EGFR signaling and HDACs, thereby reducing colon cancer cell proliferation. These findings highlight miR-134-5p as a promising therapeutic target for miRNA-mediated anticancer therapy.

结肠癌是全球癌症相关死亡的主要原因。MicroRNAs （miRNAs）是基因表达的关键调控因子，在结肠癌中经常失调。本研究旨在阐明miR-134-5p作为肿瘤抑制miRNA在结肠癌细胞中的治疗作用。我们分析了原发性和转移性结肠癌细胞中的miRNA表达谱。使用TCGA数据库评估miR-134-5p的临床意义。生物信息学工具（HADDOCK）预测了miRNA-mRNA相互作用和miRNA-mRNA- ago2复合物的分子对接。荧光素酶报告基因测定、细胞增殖、免疫荧光、集落形成单位测定和qRT-PCR分析评估了miR-134-5p对原发性和转移性结肠癌细胞中KRAS、PIK3CA和下游信号通路的影响。miR-134-5p在结肠癌细胞中下调。生物信息学分析提示KRAS、PIK3CA、EGFR和HDAC5是潜在的靶点。HADDOCK分析显示KRAS、PIK3CA、miR-134-5p和AGO2之间具有很强的结合亲和力和结构稳定性。基因报告基因检测证实了mir -134-5p介导的KRAS和PIK3CA降解。转染miR-134-5p可降低KRAS和PI3K蛋白水平，抑制EGFR/RTK信号及其下游靶点，抑制HDAC表达，最终降低结肠癌细胞增殖。本研究结果证实，miR-134-5p在结肠癌细胞中作为潜在的抑瘤miRNA，通过ago2介导的基因沉默抑制KRAS和PI3K的表达。它解除了下游EGFR信号和hdac的调控，从而减少了结肠癌细胞的增殖。这些发现强调了miR-134-5p作为mirna介导的抗癌治疗的一个有希望的治疗靶点。

{"title":"Unraveling the mechanism of microRNA-134 in colon cancer progression: Targeting KRAS and PIK3CA for cell cycle control and histone deacetylase regulation","authors":"Ganesan Jothimani , Diptimayee Das , Surajit Pathak , Sarubala Malayaperumal , Hong Zhang , Xiao-Feng Sun , Antara Banerjee","doi":"10.1016/j.yexcr.2024.114385","DOIUrl":"10.1016/j.yexcr.2024.114385","url":null,"abstract":"<div><div>Colon cancer is the leading cause of cancer-related deaths worldwide. MicroRNAs (miRNAs) are key regulators of gene expression, often dysregulated in colon cancer. This study aims to elucidate the therapeutic role of <em>miR-134-5p</em> as a tumor suppressor miRNA in colon cancer cells. We analyzed miRNA expression profiles in primary and metastatic colon cancer cells. The clinical significance of <em>miR-134-5p</em> was evaluated using the TCGA database. Bioinformatics tools (HADDOCK) predicted miRNA-mRNA interactions and the molecular docking of miRNA-mRNA-AGO2 complexes. Luciferase reporter assays, cell proliferation, immunofluorescence, colony forming unit assays, and qRT-PCR analysis assessed <em>miR-134-5p</em> effects on <em>KRAS</em>, <em>PIK3CA</em>, and downstream signaling pathways in primary and metastatic colon cancer cells. <em>miR-134-5p</em> was downregulated in colon cancer cells. Bioinformatics analysis suggested KRAS, <em>PIK3CA</em>, <em>EGFR</em>, and <em>HDAC5</em> as potential targets. HADDOCK analysis revealed strong binding affinity and structural stability between <em>KRAS</em>, <em>PIK3CA</em>, <em>miR-134-5p</em>, and AGO2. Gene-reporter assays confirmed <em>miR-134-5p</em>-mediated degradation of <em>KRAS</em> and <em>PIK3CA</em>. <em>miR-134-5p</em> transfection reduced <em>KRAS</em> and <em>PI3K</em> protein levels, suppressed <em>EGFR/RTK</em> signaling and its downstream targets, and inhibited HDAC expression, ultimately reducing colon cancer cell proliferation. The results of this study confirm that <em>miR-134-5p</em> acts as a potential tumor suppressor miRNA in colon cancer cells by inhibiting <em>KRAS</em> and <em>PI3K</em> expression through AGO2-mediated gene silencing. It deregulates downstream <em>EGFR</em> signaling and <em>HDACs</em>, thereby reducing colon cancer cell proliferation. These findings highlight <em>miR-134-5p</em> as a promising therapeutic target for miRNA-mediated anticancer therapy.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"444 2","pages":"Article 114385"},"PeriodicalIF":3.3,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142921463","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0