Cell Biochemistry and Function最新文献

4-hydroxy-2-nonenal (4HNE), a reactive aldehyde produced during lipid peroxidation, reduces angiogenesis in cultured mouse coronary endothelial cells (MCECs). In our previous studies, we found that 4HNE lowers both mRNA and protein levels of vascular endothelial growth factor receptor 2 (VEGFR2), a key regulator of angiogenesis. Since VEGFR2 transcription is regulated by nuclear factor kappa B (NFκB)—a well-known transcription factor—and aldehyde dehydrogenase 2 (ALDH2), a mitochondrial enzyme that detoxifies 4HNE, we sought to investigate how the interaction between ALDH2, 4HNE, and NFκB and VEGFR2 affects angiogenesis. Given that coronary endothelial cell rarefaction contributes to cardiometabolic conditions such as heart failure with preserved ejection fraction (HFpEF), it is imperative to study the underlying signaling mechanisms regulating coronary angiogenesis. In this study, we hypothesized that activation of ALDH2 enhances NFκB signaling, thereby preventing the 4HNE-induced reduction in VEGFR2 expression and restoring angiogenesis. To test this, MCECs were treated with disulfiram (DSF; 2.5 μM), an ALDH2 inhibitor; Alda-1 (10 μM), an ALDH2 activator; and prostratin (10 μM), an NFκB activator, prior to 4HNE (75 μM) exposure. Prostratin treatment increased both cellular and nuclear levels of NFκB across all conditions. Alda-1 pretreatment significantly rescued 4HNE-induced impairment of angiogenesis (p < 0.005 vs. 4HNE), while prostratin further enhanced Alda-1's effects (p < 0.005 vs. Alda-1 alone). Alda-1 restored VEGFR2 levels suppressed by 4HNE, and this effect was further potentiated by prostratin (p < 0.0005 vs. Alda1). In conclusion, NFκB activation enhances the protective effect of ALDH2 against 4HNE-induced angiogenic dysfunction by restoring VEGFR2 expression.

Bruton's tyrosine kinase (BTK) plays a pivotal role in intracellular signaling within B-cells, governing development, differentiation, and survival, and is an integral target in treating B-cell malignancies and autoimmune disorders. The impact on B-cell receptor, Toll-like receptor, and chemokine receptor pathways establishes its therapeutic interest. This review integrates existing studies on BTK functional roles in immunology and BTK inhibitors (BTKis) targeting. Literature was ascertained by searching prominent databases such as PubMed, Embase, Web of Science and Google scholar for BTK's molecular biology, the history of BTKis and their clinical significance. The review addresses mechanisms of resistance, particularly the C481S mutation, which impedes covalent inhibition but is abrogated by non-covalent BTKis like pirtobrutinib, including BTK's application in autoimmune and inflammatory disorders, and discusses the importance of further long-term safety and efficacy in chronic diseases. BTK inhibitors have proven to be an essential part of targeted therapy for B-cell cancer, and continuing advances are addressing challenges like drug resistance and side effects. However, major challenges like the emergence of novel resistance mutations, the need for accurate biomarkers to direct therapy, and the continuing challenge of controlling drug-associated toxicities and maintaining costs under control still pertain. Ongoing research is essential to fully realize the therapeutic potential of BTK inhibitors and integrate them into personalized regimens for cancer and immune diseases alike.

Adipose-derived stromal/stem cells (ASCs) are a readily accessible mesenchymal stem cell population with high regenerative and immunomodulatory potential. Despite their regenerative potential, ASCs experience replicative senescence during in vitro expansion, resulting in oxidative damage and impaired cellular function. Alpha-lipoic acid (ALA) is a potent antioxidant with reported anti-aging and cytoprotective effects in various cell types. This study aimed to investigate the effects of ALA on proliferation, migration, invasion capacity, and oxidative stress response of primary human ASCs in vitro. Human ASCs were isolated from adipose tissue samples and characterized via flow cytometry and multilineage differentiation. The safe and effective concentration of ALA was determined using WST-1 cell viability assays. Functional evaluations included wound healing and invasion assays. Gene expression of stemness, proliferation, cell cycle, migration, and apoptosis markers was analyzed by qRT-PCR, while MMP2 and MMP9 protein expression was assessed by immunofluorescence. Cellular oxidative stress levels were measured using DCF-DA flow cytometry, and cytotoxicity was evaluated with the LDH release assay. ALA supplementation significantly enhanced ASC proliferation (p < 0.01), migration (p < 0.001), and invasion (p < 0.01). Upregulation of MMP2 (p < 0.001) and MMP9 (p < 0.001) proteins was confirmed by immunofluorescence. ROS levels (p < 0.001) and LDH release (p < 0.001) were markedly reduced in ALA-treated cells under oxidative challenge. The ALA group also maintained higher expression of key proliferation and stemness-associated genes. ALA improves the in vitro expansion and functional properties of ASCs by enhancing proliferative and migratory capacities and mitigating oxidative stress-induced damage. These findings suggest that ALA may serve as a beneficial supplement in ASC-based regenerative applications and long-term culture systems.

TMEM63 is a mechanosensitive transmembrane protein implicated in calcium regulation; however, its specific function in cochlear auditory cells remains undefined. Calcium signaling is essential for cochlear hair cell proliferation, apoptosis, and migration—processes fundamental to hearing preservation and potential regeneration. This research sought to examine TMEM63's regulatory function in proliferation, apoptosis, and migration within HEI-OC1 cochlear hair cell systems, aimed at delineating the involvement of calcium-dependent signaling pathways, particularly the CDC42/AMPK axis. HEI-OC1 cells were subjected to TMEM63 overexpression or knockdown via plasmid transfection or RNA interference. Calcium chelation was performed using BAPTA-AM. Cell proliferation was assessed by CCK-8 screen, apoptosis via Annexin V-fluorescein isothiocyanate/propidium iodide analysis, as well as cellular cycling by flow cytometric evaluation. Wound repair combined with transwell analysis was conducted toward evaluate motility and invasiveness. Intracellular calcium levels were measured using ELISA, and signaling pathway activation was analyzed via Western blot and immunofluorescence. TMEM63 overexpression significantly promoted cell proliferation, enhanced migration, and suppressed apoptosis, while knockdown had the opposite effects. These phenotypes were reversed by BAPTA-AM, indicating a calcium-dependent mechanism. TMEM63 modulated intracellular Ca²⁺ levels and activated CDC42 and CaMKII while suppressing AMPK phosphorylation. Immunofluorescence confirmed consistent changes in CDC42, β-catenin, AMPK, and E-cadherin expression. TMEM63 regulates cochlear hair cell proliferation, apoptosis, and migration through calcium-dependent activation of the CDC42/AMPK signaling pathway. These findings highlight TMEM63 as a potential modulator of cochlear cell function and a novel target for hearing preservation strategies.

Breast cancer continues to be the most commonly diagnosed cancer among women worldwide, with estrogen receptor-positive (ER +) types making up the largest portion. At the heart of this subtype is the estrogen signaling pathway, especially the estrogen receptor alpha (ERα), which plays a major role in the development, growth, and response to these cancers. This review takes a close look at the structure and function of estrogen receptors and how they influence cancer progression. We explore current treatment strategies—including well-known drugs such as Tamoxifen and Fulvestrant, as well as aromatase inhibitors—and explain how these therapies work and why resistance sometimes develops. This review also dives into newer, more targeted options, such as HER2 inhibitors, CDK4/6 blockers, and drugs that interfere with the PI3K/AKT/mTOR pathway. These treatments are changing the game for many patients. In addition, the review highlights exciting progress in drug design, showing how researchers are improving the precision and effectiveness of cancer medications through innovations in medicinal chemistry. Antibody-drug conjugates (ADCs) are being developed to deliver powerful drugs directly to cancer cells with fewer side effects. This article also looks at emerging approaches, like oral estrogen receptor degraders (SERDs), combination therapies, and precision medicine techniques that tailor treatment based on each patient's unique genetic profile. Altogether, these developments represent a major step forward in our understanding and treatment of ER+ breast cancer.

Rho GTPases, including RhoA, Rac1, and Cdc42, are central regulators of cytoskeletal dynamics, cell migration, proliferation, and survival, and their dysregulation is firmly implicated in cancer initiation and progression. Although extensive mechanistic and translational studies have been conducted over the past three decades, the overall research landscape linking Rho GTPase signalling to cancer remains dispersed across biological processes, tumour types, and therapeutic strategies. In this study, we performed a bibliometric analysis of 1,457 articles published between 1990 and 2024, retrieved from the Web of Science Core Collection, and refined through manual screening. Quantitative analyses were conducted using Microsoft Excel, and collaboration, co-citation, and keyword networks were visualized using VOSviewer. Our analysis reveals a substantial increase in publications after 2004, with Oncogene identified as a leading journal and the United States as the most prolific contributing country. The literature predominantly focuses on core hallmarks of cancer, including epithelial-mesenchymal transition, tumour invasion, and metastasis, with breast cancer emerging as the most frequently studied disease context. Canonical Rho GTPases, RhoA, Rac1, and Cdc42 dominate the field as key mechanistic drivers and prognostic markers. In contrast, atypical Rho GTPases remain comparatively understudied. Although Rho GTPase signalling has been widely investigated as a therapeutic target, clinical translation remains challenging, largely due to signalling redundancy, pathway plasticity, and tumour adaptive responses rather than insufficient research activity. Collectively, these findings highlight persistent gaps, including limited integration with immuno-oncology and precision medicine frameworks and underexplored roles of non-canonical Rho GTPases. This bibliometric assessment provides a structured overview of three decades of research and identifies priorities to guide future investigations into Rho GTPase-driven cancer biology and therapy.

Gene therapy, utilizing a protein-based delivery system, offers a promising alternative to viral vectors owing to their stability, cell specificity, and adaptability. This study investigates the potential of collagen, a key extracellular matrix (ECM) component, as a gene delivery vehicle. While natural collagens play a crucial role in tissue regeneration, their high molecular weight limit efficient cellular uptake. To overcome this, we engineered a collagen-like protein (CLP) containing G-X-Y repeats mimicking type I collagen and retaining its functional properties. Furthermore, mussel adhesive protein (MAP) with histone-like characteristics is capable of delivering foreign genetic material into mammalian cells. We hypothesize that fusing CLP with MAP, would synergize the ECM-binding of collagen with the cationic property of MAP, enhancing gene delivery efficiency. The recombinant fusion protein was successfully expressed in E. coli and purified. The fusion protein exhibited strong DNA binding ability similar to MAP and the protein-DNA complex remained stable even in the presence of serum and DNase enzyme. The in vitro transfection studies substantiate the potential of CLP-MAP to deliver plasmid DNA in NIH-3T3 cells. Our findings indicate that the CLP-MAP fusion protein is effective in DNA binding and biocompatible, positioning it as a promising novel gene delivery system.

Food-derived bioactive peptides are gaining significant attention as safer, natural alternatives to synthetic drugs for managing hypertension. While research has predominantly focused on angiotensin-converting enzyme (ACE) inhibition, the frequent disconnect between in vitro potency and in vivo efficacy suggests the involvement of alternative molecular mechanisms. This review critically assesses recent literature regarding two pivotal non-ACE inhibitory pathways: the inhibition of renin, the rate-limiting enzyme of the renin-angiotensin system (RAS), and the upregulation of endothelial nitric oxide synthase (eNOS). We examine insights from studies characterizing peptides acting through these pathways and discuss emerging mechanisms such as angiotensin II receptor blockade and the ACE2/Ang(1-7)/MasR axis. Furthermore, we identify critical knowledge gaps arising from methodological inconsistencies in in vivo assays and the limited number of in vivo validation studies. The evidence suggests that antihypertensive food peptides function through multifaceted mechanisms, offering a holistic approach to blood pressure regulation. Future research must prioritize integrated approaches combining in silico, in vivo, and in vivo models to bridge existing translational gaps.