Biochemical Genetics最新文献

Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease linked to aging. Mitochondrial dysfunction in circulating T cells, often caused by disruption of mitochondrial DNA (mtDNA), may play a role in age-related conditions like IPF. In our previous study, we found high mtDNA mutational loads in blood lymphocytes from IPF patients, especially in regions critical for mtDNA expression. Since Complex I of the electron transport chain, partly encoded by mtDNA, is essential for energy production, we conducted a preliminary study on its activity. We found significantly reduced Complex I activity (p < 0.001) in lymphocytes from 40 IPF patients compared to 40 controls, which was positively correlated with lung function decline, specifically in functional vital capacity and diffusing capacity for carbon monoxide. These findings indicate that T cell mitochondrial dysfunction is associated with disease progression in IPF. Future work will explore the mechanisms linking T cell mitochondrial disruption with fibrosis, potentially uncovering new therapeutic targets.

Breast cancer is a heterogeneous tumor with 4 major molecular subtypes. Hormone receptor (HR)-positive and HER2-negative breast cancer accounts for 70% of invasive breast cancers. In our study, we collected 15 original Luminal B breast cancer tissue (LBBC) and paired non-cancerous adjacent tissue (NATs) from patients and performed LC-MS/MS-based label-free quantitative phosphoproteomic analysis. The untargeted metabolomics analysis was also used to determine the differences in metabolic patterns between LBBC and NATs. In addition, an integrative analysis of phosphoproteomics and metabolomics data was performed to investigate regulatory metabolic pathways. The main regulatory proteins were verified by western blot. Phosphoproteomics analysis identified 1385 differentially phosphorylated sites in 785 proteins. The protein kinase A (PKA) and protein kinase C (PKC) families and p70 ribosomal S6 kinase (RPS6K) were strongly activated in LBBC, whereas the cycle-dependent kinases (CDKs) were markedly inhibited. Cancer-specific activation of PI3K-mTORC and Hippo signaling pathways were also highlighted. Metabolomic analysis showed that 223 metabolites were significantly differentially accumulated, including fatty acids (3-hydroxycapric acid; dodecanoic acid; linoleic acid; stearic acid), glycerophospholipids, glycerophosphatidylcholines, and sphingolipids, which were mainly involved in fatty acid oxidation metabolism, sphingolipid metabolism, purine metabolism, and amino acid metabolism pathway. After integrative analysis, we found that the sphingolipid metabolic pathway played the major regulatory role. We also validated 3 phosphorylated proteins (p-YAP, p-SGK1, and p-SGPP2) in the PI3K-mTORC, Hippo signaling pathway, and sphingolipid metabolic pathway, respectively. The present study provides the first integrative phosphoproteome and metabolome profiles of LBBC, mainly involving dysregulation of sphingolipid homeostasis mediated by PI3K-mTORC and Hippo signaling pathways. This study described two phosphorylation pathways and sphingolipid metabolism regulation module for a better understanding of LBBC carcinogenesis and therapy.

Dysregulation of amino acid metabolism is recognized to have a substantial influence on tumorigenesis and the modulation of tumor microenvironment. However, the role of amino acid metabolism-related genes in diffuse large B-cell lymphoma (DLBCL) remains undefined. Therefore, we aimed to explore the influence of amino acid metabolism-related genes in DLBCL using bioinformatics approaches. Consensus clustering demonstrated that the reprogramming of amino acid metabolism has prognostic value in DLBCL. Subsequently, we developed a risk model using LASSO-Cox regression analysis to accurately predict DLBCL prognosis and identified kynureninase (KYNU) as a potentially valuable biomarker. Analysis of immune infiltration was conducted to examine the correlation between risk scores and immune profiles. Furthermore, RT-qPCR showed that the KYNU mRNA levels were upregulated in OCI-LY1, OCI-LY3, and OCI-LY10 DLBCL cells compared with normal CD19+B lymphocytes. Cell proliferation assays and flow cytometry analysis showed that inhibition of KYNU expression reduced cell proliferation and induced apoptosis of DLBCL cells. Overall, we demonstrated the significant impact of amino acid metabolism on DLBCL. Our findings may help improve the assessment of disease prognosis and provide potential therapeutic strategies for DLBCL.

Non-small cell lung cancer (NSCLC) is the leading cause of cancer death worldwide. Geniposide, an active compound of Gardeniae Fructus, has antithrombotic, antitumor, neuroprotective, hepatoprotective, cholestatic, and other effects. The present study aimed to investigate the effects of geniposide on NSCLC cells, as well as its underlying mechanism. Two NSCLC cell lines (H1975 and A549) were treated with different doses of geniposide. The proliferation, apoptosis, migratory and invasive capacities, epithelial-mesenchymal transition (EMT), and stem cell characteristics of NSCLC cells were evaluated using a series of in vitro experiments, including colony formation, flow cytometry, wound healing, transwell, western blotting, and tube formations assays. H1975 cells were subcutaneously injected into nude mice to establish the xenograft tumor models, and the models were intraperitoneally injected with 100 mg/kg geniposide or/and 6 mg/kg SKL2001, an agonist of Wnt pathway. Immunohistochemistry, immunofluorescence, and western blotting analyses of the tumors were performed. Geniposide restrained the proliferation of NSCLC cells, as shown by reduced number of colonies and downregulation of Ki67 and PCNA expression levels. Geniposide promoted apoptosis by reducing Bcl-2 expression and increasing Bax expression. Additionally, geniposide inhibited the migratory and invasive abilities of NSCLC cells as well as reversed the EMT by downregulating vimentin, N-cadherin, snail, and slug and upregulating E-cadherin in the absence or presence of TGF-β1. Furthermore, geniposide attenuated the stem cell characteristics of NSCLC cells. In mechanism, geniposide repressed the activation of Wnt/β-catenin pathway. SKL2001 reversed the anti-NSCLC effects of geniposide in vitro. In the xenograft tumor models, 100 mg/kg geniposide suppressed NSCLC tumor growth, which was reversed by SKL2001 treatment. Overall, geniposide inhibits NSCLC progression by reducing cancer cell proliferation, migration, invasiveness, EMT, and stem cell characteristics. This information might provide novel insights into the potential use of geniposide in lung cancer intervention.

Lung adenocarcinoma (LUAD) is characterized by its aggressive nature and resistance to treatment. FAM107A is a tumor suppressor gene that has been found to possess inhibitory effects in several cancers, but its role in LUAD remains unclear. This study investigated the role of FAM107A in regulating LUAD cell growth, invasion and aerobic glycolysis and also investigated the potential underlying mechanisms. Our findings revealed that FAM107A is significantly downregulated in LUAD, and its overexpression inhibited LUAD cell growth and invasion. Furthermore, FAM107A overexpression suppressed the anaerobic phase of carbohydrate metabolism in LUAD cells. Mechanistically, FAM107A regulated the CRYAB/PI3K/AKT signaling pathway, thereby inhibiting tumor progression, and similar findings were confirmed in our in vivo mouse model. In conclusion, FAM107A can suppress LUAD progression by regulating the CRYAB/PI3K/AKT pathway and aerobic glycolysis, indicating its potential as therapeutic target for LUAD.

In the past decade, the treatment of thyroid cancer (TC) has been brought to a new era, but tumor metastasis still is an intractable difficulty in clinical. LncRNA ABHD11-AS1 has been confirmed to be involved in TC progression. However, its specific mechanism remains unknown. Tissues from TC patients and TC cells served as mainly experimental subjects in this study. The migration of TC cells was assessed using the scratch assay, and the ability of cell invasion was evaluated by transwell assay. RT-qPCR and western blot were conducted to determine the levels of related genes and proteins. The dual-luciferase reporter assay was used to validate the relationships among ABHD11-AS1, miR-876-5p and CALM2. ABHD11-AS1 and CALM2 are elevated in TC cancer samples and cells, while the expression of miR-876-5p is reduced. Subsequently, the ability of TC cells to migrate, invade and EMT was inhibited by both ABHD11-AS1 knockdown or miR-876-5p overexpression. Moreover, the suppression of malignant characteristics in TC cells resulting from ABHD11-AS knockdown was counteracted by the inhibition of miR-876-5p or the upregulation of CALM2. On the mechanism, ABHD11-AS1 elevated CALM2 and promoted the malignant development of TC cells by acting as a miR-876-5p sponge. ABHD11-AS1 mediated the miR-876-5p/CALM2 axis to drive the metastasis of thyroid cancer.

Maladaptive changes in gene expression at transcriptional level in dorsal root ganglia (DRGs) after nerve injury are critical for neuropathic pain genesis. Emerging evidence reveals the important role of non-coding RNAs (ncRNAs) in regulating gene transcription. Recent studies also have showed the contribution of ncRNAs to neuropathic pain. However, the expression profile of ncRNAs in the DRGs and potential regulatory mechanism in peripheral nerve injury-induced neuropathic pain are not fully clear. We used bCCI neuropathic pain model induced by chronic constriction injury of bilateral sciatic nerves to study the expression profile and potential functional mechanism of micro RNAs (miRNAs), long non-coding RNAs (lncRNAs), circular RNAs (circRNAs) and messenger RNA (mRNA) in the DRGs by RNA sequencing and bioinformatics analysis. A total of 47 miRNAs, 337 lncRNAs, 32 circRNAs, and 2269 mRNAs were differentially expressed (DE) in the DRGs of CCI mice 14 days after surgery. KEGG analysis demonstrated nociception-related signaling pathways were significantly enriched for DEncRNAs, including Rap1, Ras, and Hippo signaling pathway. GO analysis showed neuron related biological process, membrane related cell components, and binding related molecular functions were significantly enriched. The competing endogenous RNA (ceRNA) regulatory network of DEmiRNA-DEmRNA, DElncRNA-DEmRNA, and DEcircRNA-DEmiRNA existed in the DRGs of mice with neuropathic pain induced by peripheral nerve injury. In addition, 81 pain-related DE genes had protein-protein interactions (PPI) with each other. Our findings indicated that ncRNAs are involved in the development of peripheral nerve injury-induced neuropathic pain. DEncRNAs may provide us with a new perspective in chronic neuropathic pain research and may become a potential target for pain treatment.

Lens Epithelial Cells (LECs) apoptosis is a critical driving factor of age-related cataract (ARC), but the specific molecular mechanisms remain undefined. Herein, a novel target of ROR1 regulation was identified, the mechanism was elucidated by which ROR1 and its associated pathway proteins influence hydrogen peroxide (H₂O₂)-induced apoptosis of LECs in ARC. We found decreased ROR1 expression in human cataract lens capsules compared to normal ones, the trend was also observed in young and old mice. Experiments including CCK8, Hoechst 33,342 staining, and Western blot analysis confirmed that reduced ROR1 levels were linked to H₂O₂-induced apoptosis in HLEB3 cells. To investigate its effects on cell viability and apoptosis, we created a ROR1 interference plasmid and an overexpression plasmid. The overexpression of ROR1 effectively inhibited H₂O₂-induced apoptosis of HLEB3 cells while ROR1 knockdown lowered the viability and increased the apoptosis of HLEB3 cells. Additionally, increased P38 phosphorylation was identified as a contributor to lens epithelial cell apoptosis and ARC, with ROR1 influencing this through the phosphorylation of the P38. Similarly, the relationships between P300 and STAT3, upstream of ROR1, in apoptosis of LECs and ARC were explored, and it was found that P300 and STAT3 were negatively correlated with apoptosis of LECs and ARC. In addition, the double luciferase report showed that P300 and STAT3 synergistically up-regulated the expression of ROR1. Overall, this study demonstrates that the STAT3/ROR1/P38 pathway mitigates apoptosis of LECs in ARC progression, offering a novel strategy for ARC prevention and treatment in clinical settings.

Musashi-1 (MSI1) has been proposed as a potential prognostic biomarker in prostate cancer (PCa), but its role and underlying molecular mechanisms in PCa progression remain unclear. The mRNA and protein levels of MSI1 and α/β-hydrolase domain 2 (ABHD2) in PCa tissues and cells were examined using qRT-PCR and western blot. Cell proliferation, cycle, apoptosis, and migration were detected by EdU assay, flow cytometry and transwell assay. Glucose uptake and lactate production were assessed to measure cell glycolysis. The interaction between SP1 and PLA2G6 was evaluated using dual-luciferase reporter assay and ChIP assay. MSI1 had increased expression in PCa tissues and cells. MSI1 downregulation could repress PCa cell proliferation, cycle, migration, glycolysis, and enhanced apoptosis. ABHD2 was upregulated in PCa tissues and cells, and MSI1 could bind to ABHD2 promoter region to increase its expression. Knockdown of ABHD2 suppressed PCa cell growth, migration and glycolysis, and ABHD2 overexpression also abolished the effect of MSI1 downregulation on PCa cell progression. Furthermore, interference of MSI1 reduced PCa tumor growth by decreasing ABHD2 expression in vivo. MSI1 facilitated PCa cell proliferation, migration and glycolysis via activating ABHD2 transcription, providing a novel target for PCa treatment.