Experimental cell research最新文献_第5页

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。

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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。

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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。

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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。

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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。

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引用次数: 0

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

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114320

Jiemei Shi , Min Liu , Haohao Zhu , Chunhui Jiang

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

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

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引用次数: 0

Downregulation of CTRP1 reduces radio-resistance in glioblastoma cells by inhibiting the expression of CD133 after X-ray and carbon ion irradiation 下调 CTRP1 可抑制 CD133 在 X 射线和碳离子照射后的表达，从而降低胶质母细胞瘤细胞的放射抗性

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114292

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

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

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

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引用次数: 0

lncRNA-NEF regulates hepatic stellate cells proliferation, cell cycle, apoptosis and ECM synthesis through the ERK1/2/c-Fos axis lncRNA-NEF通过ERK1/2/c-Fos轴调控肝星状细胞增殖、细胞周期、凋亡和ECM合成。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114361

Gang-gang Jia , Li-xia Lu , Bin- Li , Chu-yi Li , Ying- Zheng , Jiu-cong Zhang , Yu-jing He , Xu-Shi , Xiao-hui Yu

In this study, we investigated the role of lncRNA-NEF in modulating hepatic stellate cell (HSC) activation, a key process in liver fibrosis. Using the GSE78160 dataset, we identified lncRNA-NEF as downregulated in liver cirrhosis patients. Gene Ontology and KEGG analyses implicated it in transcriptional regulation and cell cycle control. We established an activated HSC model with TGF-β1-treated LX-2 cells and employed RT-qPCR and Western blot to assess lncRNA-NEF and ERK1/2 expression. Lentiviral transfection was used to overexpress lncRNA-NEF in activated LX-2 cells, and its effects on proliferation, apoptosis, and cell cycle were evaluated using EdU staining, CCK-8, Annexin-V PE/7-AAD, TUNEL, and PI-FACS analysis. Overexpression of lncRNA-NEF led to reduced cell proliferation, increased apoptosis, and cell cycle arrest at the S and G2/M phases. We also observed a decrease in ERK1/2, c-Fos, Collagen I, α-SMA, and Bcl-2 expression, and an increase in Caspase-3 expression, as confirmed by Western blot. These results suggest that lncRNA-NEF regulates HSC activation via the ERK1/2/c-Fos axis, potentially offering a therapeutic target for antifibrotic drug development. Our findings provide a molecular basis for understanding the role of lncRNAs in liver fibrosis and highlight the potential of lncRNA-NEF as a novel antifibrotic target.

在这项研究中，我们研究了lncRNA-NEF在调节肝星状细胞（HSC）激活中的作用，这是肝纤维化的一个关键过程。使用GSE78160数据集，我们发现lncRNA-NEF在肝硬化患者中下调。基因本体和KEGG分析表明它参与转录调控和细胞周期控制。我们用TGF-β1处理的LX-2细胞建立活化的HSC模型，采用RT-qPCR和Western blot检测lncRNA-NEF和ERK1/2的表达。采用慢病毒转染法在活化的LX-2细胞中过表达lncRNA-NEF，通过EdU染色、CCK-8、Annexin-V PE/7-AAD、TUNEL和PI-FACS分析评估lncRNA-NEF对细胞增殖、凋亡和细胞周期的影响。lncRNA-NEF过表达导致细胞增殖减少，凋亡增加，细胞周期阻滞在S期和G2/M期。我们还观察到ERK1/2、c-Fos、Collagen I、α-SMA和Bcl-2的表达减少，Caspase-3的表达增加，这一点经Western blot证实。这些结果表明lncRNA-NEF通过ERK1/2/c-Fos轴调控HSC活化，可能为抗纤维化药物开发提供治疗靶点。我们的研究结果为理解lncrna在肝纤维化中的作用提供了分子基础，并强调了lncRNA-NEF作为新型抗纤维化靶点的潜力。

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引用次数: 0

Tripartite motif (TRIM) proteins roles in the regulation of immune system responses: Focus on autoimmune diseases 三方基序蛋白（TRIM）在调节免疫系统反应中的作用：关注自身免疫性疾病。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114379

Subasini Uthirapathy , Abdulrahman T. Ahmed , Mahmood Jawad , Vicky Jain , Suhas Ballal , Hussein Riyadh Abdul Kareem Al-Hetty , Gaurav Khandelwal , Renu Arya , Muthena kariem , Yasser Fakri Mustafa

The tripartite motif (TRIM) proteins are well-studied as essential modulators of many processes, including the modulation of several pathways linked to immunological reactions. Most TRIM family members can polyubiquitinate the targeted proteins by acting as E3 ubiquitin ligases. According to current research, TRIMs play a critical role in innate immune response via modifying transcription factors, pattern recognition receptors (PRRs), and key adaptor proteins within innate immunity. It is becoming clearer that TRIMs play important roles in adaptive immune response, especially in the stimulation and promotion of T cells. We highlight the E3 ubiquitin ligase functions of TRIMs in the PRRs axis linked to autoimmune disorders. By focusing on TRIM family members, we also clarify the new approaches to regulating immunological reactions to alleviate autoimmunity.

三方基序（TRIM）蛋白是许多过程的重要调节因子，包括对与免疫反应相关的几种途径的调节。大多数 TRIM 家族成员都能通过作为 E3 泛素连接酶对目标蛋白质进行多泛素化。根据目前的研究，TRIMs 通过修饰转录因子、模式识别受体（PRR）和先天性免疫中的关键适配蛋白，在先天性免疫反应中发挥着关键作用。越来越清楚的是，TRIMs 在适应性免疫反应中发挥着重要作用，尤其是在刺激和促进 T 细胞方面。我们强调了 TRIMs 在与自身免疫疾病相关的 PRRs 轴中的 E3 泛素连接酶功能。通过聚焦 TRIM 家族成员，我们还阐明了调节免疫反应以缓解自身免疫疾病的新方法。

引用次数: 0

Reduced elastin in multiple myeloma niche promotes cell proliferation 多发性骨髓瘤小生境弹性蛋白减少促进细胞增殖。

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research

Pub Date : 2025-01-15 DOI: 10.1016/j.yexcr.2024.114395

Mozayan Zoabi , Elina Orbuch , Oded Komemi , Osnat Jarchowsky-Dolberg , Yaron Shraga Brin , Shelly Tartakover-Matalon , Metsada Pasmanik-Chor , Michael Lishner , Liat Drucker

Multiple myeloma (MM) malignant plasma cells accumulate in the bone marrow (BM) where their interactions with the microenvironment promote disease progression and drug resistance. Previously, we have shown that bone marrow mesenchymal stem cells (BM-MSCs) (MM and normal donors- ND) derived extracellular matrix (ECM) affected MM cell lines differentially with a pro-MM effect attributed to MM-MSCs' ECM. Here we studied the composition of BM-MSC's ECM (ND versus MM) with focus on elastin (ELN). Isolated BM-MSCs' ECM mass spectrometry (proteomics) demonstrated distinct differences in proteins repertoire in a source dependent manner (MM or ND-MSCs) with ELN being the most significantly decreased protein in MM-MSCs ECM. To study this observation, we cultured MM cell lines (MM1S, RPMI-8226) and BM-MSCs with/without ELN and assayed the cells' phenotype. We demonstrated that supplementing ELN to MM cell lines reduced live cell counts and increased cell adhesion. ELN also decreased MM-MSCs' proliferation but did not affect ND-MSCs. Importantly, ELN addition to MM-MSC ECM abrogated its pro-MM effect on MM cells' proliferation. These novel findings underscore a suppressive role for ELN in MM and suggest it may hold potential diagnostic and therapeutic purposes.

多发性骨髓瘤（MM）恶性浆细胞在骨髓（BM）中积累，它们与微环境的相互作用促进了疾病的进展和耐药性。先前，我们已经证明骨髓间充质干细胞（BM-MSCs）（MM和正常供体- ND）衍生的细胞外基质（ECM）对MM细胞系的影响不同，MM- mscs的ECM导致了促MM效应。在这里，我们研究了BM-MSC的ECM的组成（ND与MM），重点是弹性蛋白（ELN）。分离的MM- mscs的ECM质谱（蛋白质组学）显示，在来源依赖的方式下，蛋白质库存在明显差异（MM或ND-MSCs），其中ELN是MM- mscs ECM中减少最显著的蛋白质。为了研究这一现象，我们培养了MM细胞系（MM1S, rpm -8226）和BM-MSCs，并检测了细胞的表型。我们证明，向MM细胞系补充ELN可减少活细胞计数并增加细胞粘附。ELN也能抑制MM-MSCs的增殖，但对ND-MSCs没有影响。重要的是，在MM- msc ECM中添加ELN可消除其对MM细胞增殖的促MM作用。这些新发现强调了ELN在MM中的抑制作用，并表明它可能具有潜在的诊断和治疗目的。

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