Cytotechnology最新文献

Glycation of low-density lipoprotein (LDL) occurs when blood glucose levels are high, as in diabetic patients. This can lead to abnormal cholesterol transport in the body. Glycated LDL (G-LDL) is harmful and triggers oxidative stress in human erythrocytes. The present study investigated the protective effect of crocin, found in saffron, against G-LDL-induced cytotoxicity, oxidative damage, and formation of reactive species in human erythrocytes. These parameters were assessed in isolated human erythrocytes exposed to 6 mg/ml G-LDL, with and without varying concentrations of crocin (0.5, 1.0, 1.5 mM). Increased hemolysis, methemoglobin, and oxoferrylHb were seen in G-LDL alone-incubated cells. A significant increase in reactive species in G-LDL-exposed erythrocytes led to enhanced oxidation of lipids, proteins, and thiols. The activities of certain key antioxidant and membrane-bound enzymes were reduced. The antioxidant capacity of cells was compromised as indicated by a diminished ability to neutralize free radicals and donate electrons. G-LDL significantly altered surface morphology, forming echinocytes and agglutinating the cells. All these characteristics were significantly restored when erythrocytes were pre-treated with crocin, before incubation with G-LDL, in a crocin concentration-dependent manner. Furthermore, erythrocytes incubated with 1.5 mM crocin alone did not show alterations in any of the above parameters, indicating that crocin was not toxic to these cells. These results clearly show that crocin is strongly cytoprotective against G-LDL-induced damage and toxicity in erythrocytes. Hence, it can be used as an efficient dietary antioxidant in various atherosclerotic cardiovascular disorders, as seen in diabetic patients.

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Oral squamous cell carcinoma (OSCC) is a common malignant tumor of the head and neck, representing a significant public health concern. Neurexophilin-4 (NXPH4) facilitates cancer cell proliferation and invasion. E26 oncogene homolog 1 (ETS1) can influence OSCC cell migration and invasion by modulating various factors. This study aims to explore the molecular mechanism of NXPH4 and ETS1 in the progression of OSCC. Bioinformatics analysis was used to analyze the NXPH4 and ETS1 expression in OSCC tissues and explore the relationship between NXPH4 and tumor stage. The mRNA and protein levels were examined by reverse transcription-quantitative polymerase chain reaction (qRT-PCR) and western blot. Besides, the OSCC tumor models were constructed, and tumor volume and weight were detected. The immunohistochemistry (IHC) assay was used to analyze Ki-67 expression in vivo. The apoptosis and reactive oxygen species (ROS) were measured by flow cytometry. The levels of malondialdehyde (MDA) and ferrous ion (Fe²⁺) were detected using the corresponding detection kits. The cell proliferation was tested by 5‑ethynyl‑2'‑deoxyuridine (EdU) staining. Tumor Immune Estimation Resource (TIMER) 2.0 and flow cytometry were used to analyze the relationship between NXPH4 and macrophage M2 polarization. The interaction between NXPH4 and ETS1 was demonstrated by chromatin immunoprecipitation (CHIP) and a dual luciferase reporter gene assay. NXPH4 expression was significantly increased in OSCC tissues and CAL-27 and SCC-25 cell lines (P < 0.01). Silencing NXPH4 inhibited the OSCC tumor growth in vivo. In human OSCC cell lines, silencing NXPH4 promoted the apoptosis, MDA, and ROS levels and suppressed the GPX4 expression (P < 0.01), cell proliferation, and macrophage M2 polarization. Furthermore, ETS1 could bind to NXPH4 and positively regulate its expression. Knockdown of ETS1 facilitated the apoptosis, increased MDA and ROS levels, and repressed the GPX4 expression, cell proliferation, and macrophage M2 polarization. However, up-regulation of NXPH4 in vitro reversed the effect of sh-ETS1 in human OSCC cell lines. ETS1 elevates the progression of OSCC and enhances macrophage M2 polarization by contributing to NXPH4. These findings identify the ETS1/NXPH4 axis as a potential therapeutic target in OSCC.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00869-6.

Non-small cell lung cancer (NSCLC) severely impairs patients' health and quality of life, placing a substantial burden on both patients' families and society. N4-acetylcytidine (ac4C), a pivotal RNA modification among the most abundant in eukaryotes, regulates disease development. Research has shown the ac4C-related gene NAT10 promotes lung cancer progression, yet the mechanism is unclear. NAT10 expression was assessed using data from the TCGA, CPTAC, ENCORI, and TNMplot databases, as well as qRT-PCR, and Western blot analyses. Cell proliferation was evaluated using MTT and EdU assays. Apoptosis was detected using flow cytomety. Cell invasion and migration were respectively detected by transwell and wound-healing assays. Cancer stem cell properties were analyzed via tumor sphere formation assays. Moreover, dual-luciferase reporter assay, ChIP assay, and RIP assay were performed to analyze the interaction between CEBPA and NAT10. Additionally, mRNA stability was evaluated using actinomycin D assay. The animal model was established to analyze the effects of NAT10 and CEBPA on tumor growth in vivo. NAT10 showed high expression levels in NSCLC tissues and cells. Silencing NAT10 inhibited cell viability, proliferation, invasion, migration, and stemness, while promoting apoptosis in NSCLC cells. The transcription factor CEBPA could bind to NAT10, thereby promoting NAT10 expression. Notably, NAT10 mediated the ac4C modification of CEBPA mRNA, which enhanced CEBPA mRNA stability and accelerated the malignant progression of NSCLC. Finally, NAT10 upregulated CEBPA expression, which in turn promoted tumor growth in vivo. NAT10 catalyzed ac4C acetylation on CEBPA mRNA, enhancing its stability and increasing CEBPA protein expression, thereby promoting the malignant progression of NSCLC. The identification of this feedback loop provides a preclinical rationale for developing NSCLC therapeutic strategies targeting NAT10 or CEBPA.

Sepsis, a life-threatening systemic inflammatory response syndrome mediated by dysregulated host-pathogen interactions, frequently precipitates in renal injury. Renal tubular epithelial cell (RTEC) injury is a hallmark of septic acute kidney injury (AKI). Recent studies have demonstrated the involvement of serum response factor (SRF) in septic AKI. Herein, the role and mechanism of SRF as a transcription factor in regulating RTEC dysfunction were explored. HK-2 cells (the human RTEC line) were treated with lipopolysaccharide (LPS) for the establishment of in vitro septic AKI models. HK-2 cell viability was validated using CCK-8 assay. HK-2 cell apoptosis was evaluated by flow cytometry analysis. Measurement of proinflammatory cytokine concentration was conducted using enzyme-linked immunosorbent assay kits. RT-qPCR were required for determining gene levels. Western blotting was prepared for testing the protein levels of proinflammatory cytokines, apoptosis-related markers, NF-κB p65 and NF-κB inducing kinase (NIK). The binding of SRF to NIK promoter was confirmed by chromatin immunoprecipitation and luciferase reporter assays. LPS treatment suppressed HK-2 cell viability and accelerated HK-2 cell inflammation and apoptosis, which was antagonized by SRF depletion. Mechanically, SRF as a transcription factor bound to NIK promoter. SRF silencing inhibited LPS-induced NF-κB signaling activation in HK-2 cells. In rescue assays, NIK overexpression counteracted the restrictive impact of SRF deficiency on LPS-induced HK-2 cell dysfunction. SRF aggravates LPS-elicited HK-2 cell injury via binding to NIK promoter and activating NF-κB signaling.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00854-z.

Diabetic kidney disease (DKD) is a leading cause of kidney failure. It is characterized by damage to renal tubular endothelial cells, which is exacerbated by macrophage-derived exosomes under high-glucose (HG) conditions. Triptolide (TPL), an active ingredient extracted from Tripterygium wilfordii Hook F, has been used in the treatment of DKD. In this study, we aimed to investigate whether TPL protects against renal damage induced by macrophage-derived exosomes under HG conditions and to explore the involvement of the protein kinase B/mammalian target of rapamycin (AKT/mTOR) signaling pathway in this process. Exosomes were isolated from macrophages cultured in normal glucose (NG) and HG environments and characterized using transmission electron microscopy, nanoparticle tracking analysis, and Western blot. Subsequent in vitro experiments on mouse renal tubular epithelial cells (mRTECs) demonstrated that 5 ng/mL TPL enhanced cell viability, reduced apoptosis and inflammation, and promoted autophagy-effects that were reversed by the autophagy inhibitor chloroquine (CQ). Molecular docking and bioinformatic analyses suggested a stable interaction between TPL and AKT, implicating the AKT/mTOR pathway in TPL-mediated autophagy activation. Further in vivo studies using an AKT agonist supported the role of TPL in inhibiting AKT/mTOR signaling and enhancing autophagic activity, ultimately ameliorating renal injury in a DKD model. These findings provide novel insights into the mechanisms by TPL may mitigate exosome-induced renal damage, highlighting its potential as a therapeutic agent for DKD.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00883-8.

Cervical cancer (CC) represents the fourth most commonly diagnosed cancer and main cause of cancer mortality among women globally. We aim to investigate the mechanism of HOXB7 in CC cell progression, providing novel therapeutic implications for CC. Levels of HOXB7, miR-552-3p and IFITM1 in CC cells and tissues were measured by RT-qPCR and WB. After transfection of HOXB7 siRNA into CC cells, cell proliferation, invasion and migration were detected by CCK-8 method and transwell. The bindings of HOXB7 to miR-552-3p promoter and miR-552-3p to IFITM1 were analyzed. Effect of miR-552-3p and IFITM1 on the biological function of CC cells was detected in combined experiments. Results showed that the expression of HOXB7 and miR-552-3p was increased and the expression of IFITM1 was decreased. Cell proliferation, invasion and migration were reduced upon knockdown of HOXB7. Mechanically, HOXB7 bound and upregulated miR-552-3p expression, promoted the targeted binding of miR-552-3p to IFITM1, and reduced IFITM1 expression. miR-552-3p overexpression or IFITM1 downregulation reduced the inhibitory action of HOXB7 silencing on the proliferation, invasion and migration of CC cells. In conclusion, HOXB7 promotes the progression of CC cells via the miR-552-3p/IFITM1 axis.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-026-00903-1.

Microglial polarization plays a crucial role in Parkinson's disease (PD). This study explores how serpin family A member 1 (SERPINA1) suppresses neuroinflammation and alleviates neuronal damage in PD. Adeno-associated viruses were injected into mice to manipulate the expression of SERPINA1 or runt-related transcription factor 1 (RUNX1) in the substantia nigra, followed by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) modeling. Behavioral tests, histopathology (HE and Nissl staining), immunohistochemistry (IHC), immunofluorescence, and enzyme-linked immunosorbent assay were conducted to evaluate neuroinflammation and neuronal damage in mice. BV2 microglial cells were infected with lentiviruses overexpressing SERPINA1 and treated with 100 µM 1-methyl-4-phenyl-pyridinium (MPP)⁺. MPP⁺ increased pro-inflammatory cytokines and iNOS while decreasing anti-inflammatory cytokines and arginase-1 expression in BV2 cells. SERPINA1 and RUNX1 were upregulated in the SN of MPTP-induced mice. RUNX1 bound to the promoter region of SERPINA1 to induce its transcription. SERPINA1 or RUNX1 overexpression alleviated PD-related neuronal damage and neuroinflammation in mice and MPP⁺-induced inflammation in BV2 cells. SERPINA1 knockdown inhibited M2 polarization in the presence of RUNX1 overexpression. Taken together, RUNX1 transcriptionally activates SERPINA1, promoting microglial M2 polarization, suppressing neuroinflammation, and alleviating neuronal damage in PD.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00878-5.

Oral squamous cell carcinoma (OSCC) is an aggressive malignancy with poor prognosis and limited therapeutic options. Plasminogen activator, urokinase (PLAU) is emerged as a potential key player in OSCC by bioinformatics analysis, but its function in OSCC have not been explored. Therefore, this study investigates its role and mechanism in OSCC progression, with a focus on its interaction with the N6-methyladenosine (m⁶A) writer ZC3H13. Differentially expressed genes in OSCC were identified through analysis of GEO datasets. A candidate oncogene was subsequently selected based on Gene Ontology enrichment analysis and validated using qRT-PCR in patient samples. The role of PLAU was examined by CCK-8, EdU, and transwell assays. In vivo tumorigenicity was evaluated using xenograft models. The relationship amongst PLAU and ZC3H13 was analyzed through correlation analysis, MeRIP, RIP, qRT-PCR, immunoblotting and mRNA stability assays. Finally, rescue experiments were conducted to validate the interactions between ZC3H13 and PLAU. PLAU upregulated in OSCC was identified as a candidate oncogene based on bioinformatic analysis and mRNA expression profiling. PLAU silencing suppressed cell proliferation, migration, invasion, and tumor growth, whereas its overexpression produced the opposite effects. Furthermore, ZC3H13 was positively correlated with PLAU expression. ZC3H13 overexpression enhanced m⁶A levels of PLAU to increase PLAU expression and mRNA stability. In addition, ZC3H13 overexpression partially rescued the suppressive effects of PLAU silencing on OSCC cells. In summary, these results illustrate that ZC3H13-mediated m⁶A modification increases PLAU mRNA stability and expression, thereby enhancing OSCC progression.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00882-9.

Gastric cancer (GC) is one of the most common malignant tumors worldwide. Banxia Xiexin Decoction (BXXXD) has a significant therapeutic effect on digestive system diseases, but its mechanism of action on GC is not yet clear. Using single-cell and bioinformatics to discover prognostic markers and Tumor Microenvironment (TME) components for GC, and further explore the molecular mechanism of BXXXD in treating GC through network pharmacology. KIF2C, KIF20A, KIF11, CDK1, CDC20, FN1, etc. could be used as diagnostic and prognostic markers for GC. The tumor microenvironment of GC mainly includes T cell, Neutrophil, B cell, Macrophage, Epithelial cell, Plasma cell, Stromal cell, Fibroblast, common myeloid progenitor cell. The core active ingredients of BXXXD synergistically regulate the MAPK signaling pathway, oxidative stress response, and apoptosis pathway, and multi-target reverse the pathological process of GC. We have established the correlation between the pathogenesis of GC's "cold and heat disorder" and modern pathological indicators (inflammation, oxidative damage, and cell apoptosis disorder) from the perspective of traditional Chinese medicine syndrome, providing a theoretical basis for the clinical application of BXXXD.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00888-3.

The anti-proliferation, anti-invasion, and anti-migration activities of KLF2 in keloid fibroblasts (KFs) and hypertrophic scar fibroblasts (HSFs) were investigated. KLF2 in normal skin (NF), keloid (KD), hypertrophic scar (HS) tissues, and cell lines was quantified using western blot and quantitative real-time polymerase chain reaction techniques. The processes of cell proliferation, migration, and invasion were observed in both KFs and HSFs, with pathway proteins linked to these processes pinpointed through western blot analysis. KLF2 in KD and HS tissues was lower than that in NF tissues. Enhancing KLF2 expression inhibited KF and HSF proliferation, migration, and invasion. MMP2, MMP-9, PI3K and p-Akt protein levels were inhibited in KFs with KLF2 overexpression. However, inhibition of PI3K and p-Akt protein levels was observed only in KLF2-overexpressed HSFs. In KFs with enhanced KLF2 expression, PI3K agonists eliminated the effect of KLF2 overexpression on cell migration and invasion. KLF2 inhibits the proliferation and migration of KFs by down-regulating MMP-2/9 through the PI3K/AKT pathway, suggesting that KLF2 may be a potential therapeutic target for KD. Hence, these findings offer novel perspectives on the function and molecular pathways of KLF2 in KD, as well as novel strategies for its clinical treatment.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00885-6.