Human Cell最新文献

Background: SIRT1 exhibited a protective role in myocardial ischemia/reperfusion injury (MI/RI), but the related mechanisms remained unclear. In this study, the regulation of SIRT1 on neddylation modification in MI/RI was explored.

Methods: H9C2 cells underwent hypoxia and reoxygenation (H/R) to mimic MI/RI in vitro, and C57BL6 mice were employed to establish MI/RI model for the in vivo experiments. Mass spectrometry analysis was employed to screen the possible modified substrates of NEDD8; Western blot was performed to detect protein level; CCK8 was performed to assess cell viability; flow cytometry, TUNEL, and Cardiac Troponin T (cTNT) double staining were performed to assess cardiomyocytes apoptosis; TTC and HE staining were performed to assess infarction area and pathological changes of cardiac tissues in MI/RI mice, respectively.

Results: MLN4924 (an inhibitor of NEDD8-activating enzyme (NAE)) significantly reversed the elevated NEDD8 conjugated protein (p < 0.001) and reduced SIRT1 protein levels (p < 0.001) induced by H/R in H9C2 cells. Dead-box helicase 5 (DDX5) was screened as the possible modified substrate of NEDD8 via mass spectrometry. H/R further reduced DDX5 protein level (p < 0.001) and increased DDX5 neddylation in H9C2 cells, while which were reversed by MLN4924 or LV-SIRT1 (p < 0.05). Also, SIRT1 increased DDX5 protein level by enhancing DDX5 stability via reducing its neddylation. Functionally, hypoxia decreased cell viability (p < 0.001) and increased cell apoptosis (p < 0.001) and ROS level (p < 0.001) in H9C2 cells, whereas they were all reversed by LV-SIRT1 (p < 0.05, p < 0.001) or LV-DDX5 (p < 0.05, p < 0.001). The in vivo experiments revealed that LV-DDX5 reversed the increased infarction area (p < 0.05), necrotic myocardial fibers and cardiomyocytes apoptosis (p < 0.001) in MI/RI mice.

Conclusion: These results suggested that SIRT1 increased DDX5 protein level to reduce cardiomyocytes apoptosis and ROS level via the inhibition of DDX5 neddylation, thus alleviating MI/RI.

The regenerative ability of the skin is orchestrated by different types of cells, including mesenchymal stromal cells (MSCs). Their role in wound healing is being widely studied due to their capacity to produce a secretome able to modulate their microenvironment. In this context, MSCs factors can be isolated using in vitro cell culture and be experimentally tested for a series of clinical conditions, such as skin repair. In this study, we produced conditioned medium (MSC-CM) using primary cultures from human adipose-derived MSCs. This medium was used as treatment in a culture of keratinocytes cell line from adult human skin (HaCaT) to investigate the modulation of their dynamics. We analyzed cell proliferation, migration, and changes on cell morphology by labeling the actin filaments and nuclei. The factors released by MSCs were able to improve both the proliferation and migration of keratinocytes. In addition, there was an increase in the amount of actin stress fibers, filopodia protrusions, and nuclei irregularities. The MSCs secretome modified the migratory patterns of keratinocytes, being observable through their morphological changes. At least in part, this modulation was caused by the TGF-β1 signaling, considering that its antagonist, SB 431542, lead to a reduction of approximately 76% in the migration of HaCaT cells through the porous membranes of transwell chambers. Together, our data contribute to a better understanding of the role of MSCs on keratinocytes during wound healing and reinforce the importance to investigate their potential in dermal regeneration therapies.

In humans, circadian rhythm regulates the expression of numerous protein-coding genes across nearly all cell types, playing a central role in maintaining health. However, circadian rhythm components interact with a wide variety of intracellular signaling pathways. Through these interactions, the circadian system interfaces with various pathologies by governing different molecular mechanisms. Evidence suggests that the interaction between circadian rhythms and apoptosis plays a pivotal role in both health and disease, warranting careful investigation. Apoptosis, one of the main mechanisms of programmed cell death, is essential for establishing maintaining cellular homeostasis in the human body. Alterations in apoptotic activity occur during the progression of cancer, as well as in various autoimmune and neurodegenerative diseases. Conversely, bidirectional interactions between circadian components and apoptotic factors offer new insights into disease development. Therefore, this review highlights the links between circadian rhythms and apoptosis across health and disease states, aiming to identify potential therapeutic interventions based on this connection.

Various exosome-derived proteins have been reported to play essential roles in regulating colorectal cancer progression and affecting the prognosis of cancer patients. It is necessary to explore the critical exosome-related genes in colorectal cancer. In this study, 23 differentially expressed exosome-related genes associated with prognosis in colorectal cancer (CRC) were identified based on two datasets, The Cancer Genome Atlas (TCGA) and exoRbase. Based on machine learning-Boruta and lasso-Cox regression-nine essential genes were finally identified, and a risk model was constructed. The risk model was able to predict the prognosis of the patients well. Specifically, the prognosis of high-risk patients was worse, and the prognosis of low-risk patients was better. Multivariate Cox regression revealed that the risk model was an independent prognostic factor. Mechanism studies showed that pathways such as MYOGENESIS, APICAL JUNCTION, Epithelial-Mesenchymal Transition (EMT), ANGIOGENESIS, and KRAS SIGNALING DN were highly enriched in the high-risk group. In addition, tumor-promoting immune cells, such as Treg cells and macrophages, exhibited increased activity in the high-risk group, suggesting that high-risk patients may be less responsive to immunotherapy. Furthermore, in multiple external immunotherapeutic and chemotherapeutic datasets, we found that high-risk patients are less sensitive to therapy than low-risk patients, suggesting that this risk score may predict immune and chemotherapy response. Scoring the importance of nine genes, we found that TIMP1 was the most critical exosome-related gene in colorectal cancer patients. Knockdown of TIMP1 in colorectal cancer cells significantly inhibited the proliferation and migration of colorectal cancer cells. In conclusion, we identified several crucial colorectal cancer exosome-associated genes using public datasets and machine learning, and constructed risk models to predict prognosis and response to immunotherapy. TIMP1 was further identified as a critical oncogene in patients with colorectal cancer. Our results provide a theoretical basis for subsequent exosome-based preclinical trials.

Cardiac fibrosis is a prevalent pathological feature in the progression of various cardiovascular diseases, including heart failure, myocardial infarction, and dilated cardiomyopathy, particularly in their advanced stages. Its primary mechanism involves the abnormal activation of cardiac fibroblasts and excessive deposition of extracellular matrix, which ultimately results in decreased myocardial compliance and cardiac dysfunction. The Hippo signaling pathway, an evolutionary conserved kinase cascade, not only regulates organ development and tissue homeostasis but has also been shown to play a critical role in cardiac fibrosis. Notably, the Hippo pathway demonstrates cell-specific regulatory functions across different cardiac cell types, including cardiomyocytes, fibroblasts, and immune cells. This systematic review elucidates the molecular mechanisms by which the Hippo pathway influences cardiac fibrosis, emphasizing its cell type-dependent roles. It analyzes the complexity of its roles from the perspectives of cross-talk between pathways, various types of cardiac diseases, and different stages of disease progression. Additionally, it summarizes recent advancements in anti-fibrotic drugs that target this pathway, thereby providing a theoretical foundation for the development of novel therapeutic strategies in cardiac fibrosis.

Non-small cell lung cancer (NSCLC) is a leading cause of cancer-related mortality worldwide, with poor prognosis largely attributed to late-stage diagnosis and therapeutic resistance. Cisplatin (DDP) resistance is a major challenge in NSCLC treatment. Emerging evidence highlights the critical role of N6-methyladenosine (m6A) RNA modification in regulating cancer progression and drug resistance, with methyltransferase-like 3 (METTL3) serving as the key methyltransferase mediating m6A deposition on RNAs. In this study, we investigated the role of the circular RNA hsa_circ_0003176 in DDP-resistant NSCLC and its regulation by m6A modification. We found that hsa_circ_0003176 was significantly downregulated in DDP-resistant NSCLC cells and functioned as a tumor suppressor by promoting autophagy, inhibiting glycolysis, and reversing DDP resistance. Mechanistically, METTL3-mediated m6A modification suppressed hsa_circ_0003176 expression, while hsa_circ_0003176 directly targeted and destabilized ribosomal protein S6 kinase B1 (RPS6KB1) mRNA, a key regulator of mTORC1 signaling, thereby inhibiting NSCLC progression. In addition, in vivo xenograft models confirmed that hsa_circ_0003176 overexpression suppressed tumor growth and enhanced DDP sensitivity. Our study reveals the METTL3/m6A/hsa_circ_0003176/RPS6KB1 pathway as a critical pathway in NSCLC chemoresistance, offering novel therapeutic targets for overcoming DDP resistance.

Ethambutol (EMB)-induced optic neuropathy (EON) is a clinical concern. Ferroptosis, involving iron and toxic reactive oxygen species (ROS), causes unique cell death, but its mechanism in EON is unclear. This study aims to explore the EON mechanisms. Wistar rats were used to establish an EON model by administering EMB at 50 mg/kg daily for 8 weeks. Retinal ganglion cells (RGC-5 cells) were used for in vitro experiments. Histological staining, MTT assays, flow cytometry, western blot analysis, dual-luciferase reporter assay, chromatin immunoprecipitation, and high-throughput sequencing were conducted to investigate cell death modes and molecular changes. EMB treatment leads to significant cell loss and structural damage in RGCs of EON model, predominantly through ferroptosis. We confirm increased ROS levels, downregulation of SLC7A11 and GPX4, and decreased glutathione (GSH) levels, upregulation of malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) levels in EMB-treated RGC-5 cells. Furthermore, sequencing data reveal that in RGC-5 cells treated with EMB, the differentially expressed genes (DEGs) primarily exhibited alterations in biological functions associated with metabolism, stress response, and apoptotic regulation. Specifically, EMB inhibits the expression of succinate dehydrogenase enzyme B (SDHB), thereby disrupting antioxidant defenses and facilitating ferroptosis. Moreover, Smad4 has been pinpointed as a pivotal transcription factor in regulating SDHB expression. Notably, its interaction with the promoter region of SDHB is inhibited by EMB. This study provides compelling evidence for the involvement of ferroptosis in EON and highlights SDHB and Smad4 as potential therapeutic targets for mitigating this adverse effect.

Lung adenocarcinoma (LUAD), the most prevalent and aggressive form of non-small cell lung cancer. Natural compounds have gained increasing attention as potential anti-cancer agents. The therapeutic potential of chebulagic acid (CA)-a hydrolysable tannin with documented anti-proliferative properties-has not been investigated in LUAD. In the present study, functional experiments revealed that CA treatment markedly suppressed proliferation and induced mitochondrial-dependent apoptosis of LUAD cells. In addition, CA augments natural killer (NK) cell migration by enhancing LUAD CCL5 production. Quantitative proteomics identified FBXO38 as the most significantly upregulated protein following CA treatment. FBXO38 silencing abrogated CA-induced CCL5 production and NK cell migration in LUAD cells. The in vivo experiments showed that CA significantly inhibited tumor growth and enhanced NK cell infiltration in mice, accompanied by decreased Ki67⁺ proliferating cells, increased cleaved caspase-3⁺ apoptotic cells, and upregulated FBXO38 expression. Collectively, our findings demonstrate that CA may be exert anti-LUAD effects through FBXO38/CCL5-mediated NK cell recruitment.

The growth of human pulmonary arterial smooth muscle cells (hPASMCs) is one of the key contributors to vascular remodeling in pulmonary arterial hypertension (PAH). Although histone deacetylase 3 (HDAC3) has been implicated in acute lung injury and pulmonary fibrosis, its role in hypoxia-induced PAH remains unclear. Here, the function and associated mechanisms of HDAC3 in hypoxia-induced hPASMC proliferation were investigated. A hypoxia-induced hPASMC model was constructed to evaluate the role of HDAC3 in cell proliferation under hypoxic conditions. The effects of HDAC3 siRNA and ruxolitinib, a JAK pathway inhibitor, were assessed to explore the regulatory mechanism of HDAC3 in vascular remodeling. Hypoxia significantly upregulated both HDAC3 mRNA and protein. Inhibition of HDAC3 attenuated hypoxia-induced proliferation in hPASMCs. Moreover, HDAC3 inhibition downregulated CSF2 and suppressed proliferation by inactivating the JAK2/STAT5 axis. In contrast, HDAC3 overexpression enhanced CSF2 expression, activated JAK2/STAT5, and promoted hPASMCs' proliferation under hypoxia. Notably, the pro-proliferative and pathway-activating effects of HDAC3 overexpression were reversed by CSF2 silencing or ruxolitinib treatment. HDAC3 plays a key role in hypoxia-induced hPASMC dysfunction. Its inhibition mitigates aberrant proliferation through a CSF2-dependent inactivation of the JAK2/STAT5 pathway under hypoxia. These results indicate the potential of using HDAC3 for treating hypoxia-induced PAH.

Hydatidiform mole (HM) is a pathological pregnancy characterized by excessive trophoblast proliferation and the absence of embryonic tissue development, predominantly sporadic in onset. Recurrent hydatidiform mole (RHM) affects approximately 1%-4% of HM patients, among which familial RHM (FRHM) is extremely rare and classified as a monogenic autosomal recessive disorder. NLRP7 (NLR family, pyrin domain containing 7) is the major pathogenic gene for RHM, in which affected individuals have a profound impairment in fertility and a markedly elevated risk of malignant transformation. Yet mechanistic research remains constrained by ethical limitations in human embryo studies and the absence of animal models recapitulating HM phenotypes. Here, we report the generation and characterization of iPSC lines from FRHM patients harboring homozygous NLRP7 variants c.2078G > A (p.Arg693Gln) and c.2161 C > T (p.Arg721Trp), respectively. These cellular models offer a unique platform to dissect molecular pathways driving NLRP7-mediated reproductive failure, overcoming long-standing barriers in FRHM pathogenesis research.