Cytotechnology最新文献

The aggressive phenotype of colorectal cancer (CRC) is largely driven by interactions within the tumor microenvironment, specifically the co-occurrence of inflammation and hypoxic stress. While the pro-metastatic enzyme Kallikrein-related Peptidase 4 (KLK4) is known to contribute to dissemination, the precise molecular mechanism by which IL-6 and hypoxia converge to regulate KLK4 expression and subsequent metastatic potential remains to be elucidated. This study investigated the influence of the IL-6 cytokine on KLK4 gene expression and metastatic potential in the HT-29 colon cancer cell line under both normal and hypoxic conditions. Healthy non-cancerous endothelial cells (HUVEC) served as a comparative control. Expression was assessed via Real-Time PCR (mRNA) and Western blot (protein), while metastatic potential was determined by the scratch assay. Our findings demonstrate a substantial and marked upregulation in KLK4 gene and protein expression in HT-29 cells over a 48-hour period in response to IL-6, hypoxia, and the combined treatments. This increase in KLK4 was found to be associated with simultaneous upregulation of STAT-3 and p-STAT-3 proteins, strongly suggesting that the STAT-3 signaling pathway mediates this induction. The effects observed were tumor-specific: the non-cancerous HUVEC line showed only transient KLK4 changes and decreased proliferation in individual treatments. In sharp contrast, the combined IL-6 and hypoxia treatments significantly enhanced proliferative activity and metastatic potential in HT-29 cells. Western blot analysis collectively indicates that the augmented KLK4 expression in CRC cells is likely mediated through IL-6 and hypoxia-induced STAT-3 activation. These findings establish KLK4 as a potential downstream effector of the IL-6/STAT-3 pathway, offering a novel therapeutic target for mitigating metastatic potential in colon cancer.

Derived from Rehmanniae Radix, Rehmannioside A (ReA) appears to provide a defensive effect against diseases. This study was designed to investigate ReA's role and mechanism in cerebral ischemia-reperfusion injury (CI/RI). A transient middle cerebral artery occlusion (tMCAO) mouse model was developed. tMCAO mice were intraperitoneally injected with different doses of ReA for 3 days, and then underwent neurological function examination. Then, brain histopathology was observed by HE staining, neuronal apoptosis was observed by TUNEL staining, glial fibrillary acidic protein (GFAP) expression was detected by immunofluorescence staining to assess astrocyte activation, and blood-brain barrier (BBB) integrity was assessed by determining hemoglobin content and brain water content. p38 MAPK pathway-related proteins were detected by Western blot. Treatment with ReA in tMCAO mice showed a dose-dependent reduction in BBB damage, improvements in neurological function, decreased neuronal apoptosis, and inhibition of astrocyte activation. ReA inhibited p38 MAPK pathway activation, and the p38 MAPK pathway inhibitor SB203580 potentiated the ameliorative effects of ReA on CI/RI. ReA improves CI/RI by inhibiting astrocyte activation and reducing BBB damage through modulation of the p38 MAPK pathway.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00859-8.

This study investigates the characterization and biological effects of Dual Drug-Loaded Nanoparticles on HepG2/doxorubicin (DOX) cells, focusing on the anti-cancer ability of Doxorubicin/Curcumin-Polyethylene Glycol-Polycaprolactone Nanoparticles (DOX/Cur-PEG-PCL-NPs). DOX/Cur-PEG-PCL-NPs, along with single-drug and blank nanoparticles, were prepared and characterized by nanoparticle tracking analysis (NTA) for hydrodynamic diameter and Zeta potential. The morphology of DOX/Cur-PEG-PCL-NPs was characterized by transmission electron microscopy (TEM). Drug loading, encapsulation efficiency, and release profiles were evaluated. In HepG2/DOX cells, the DOX/Cur-NPs significantly reduced viability, migration, and invasion, while promoting apoptosis and reactive oxygen species (ROS) generation. Molecular analyses indicated downregulation of Nuclear Factor Erythroid 2-Related Factor 2 (Nrf2), NAD(P)H Quinone Dehydrogenase 1 (NQO1), Heme Oxygenase 1 (HO-1), and Multidrug Resistance Protein 1 (MDR1), and upregulation of Kelch-like ECH-associated protein 1 (Keap1), Bcl-2-associated X protein (Bax), and caspase-3. These results demonstrate that DOX/Cur-PEG-PCL-NPs enhance anticancer efficacy by modulating oxidative stress and apoptosis pathways. These results demonstrate that DOX/Cur-PEG-PCL-NPs effectively reverse chemoresistance and suppress tumor progression through modulation of the Nrf2 pathway and apoptosis induction, offering a promising strategy for targeted liver cancer therapy.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00855-y.

Apigenin (API) is a natural compound with an anti-cancer effect. This study aimed to investigate the mechanism of API against liver cancer. In vitro and in vivo, Huh7 cells and H22-induced liver orthotopic hepatoma model were constructed to assess the anti-tumor effect of API. Cell viability, apoptosis, mitochondrial membrane potential (MMP), reactive oxygen species (ROS), spleen lymphocyte differentiation, relative factors, and tumor pathological damage were determined using cell counting kit-8, flow cytometry, enzyme-linked immunosorbent assay, western blot, immunofluorescence, Hematoxylin-Eosin staining, and TUNEL. API inhibited liver cancer cell viability and promoted apoptosis both in vitro and in vivo. API increased ROS, interleukin (IL)-1β/6/8 and tumor necrosis factor (TNF)-α, expression of cleaved-Caspase-3/9, B-cell lymphoma-2 associated X protein, and phosphorylated nuclear transcription factor-kappa B proteins, while it reduced MMP of Huh7 cells. API increased interferon (IFN)-γ⁺ CD4⁺ cells (Th1) and decreased IL-4⁺ CD4⁺ cells (Th2) with increasing tumor TNF-α and IFN-γ and decreasing IL-1β and IL-4. The anti-tumor and T-cell regulating effects of API are similar to those of positive control cyclophosphamide. This study displayed that API has the potential to effectively prevent liver cancer by triggering mitochondrial apoptosis, thereby regulating the Th1/Th2 balance.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00868-7.

This study investigated whether the microrchidia family CW-type zinc finger 2(MORC2) decreases cellular response to paclitaxel by modulating glycolysis in ovarian cancer. In resistant ovarian cancer cell sublines for paclitaxel, including A2780/Taxol and SKOV3/Taxol, and parental cell lines, including A2780 and SKOV3, the differential expression of MORC2 was confirmed via qRT-PCR at the mRNA level and through western blot analysis at the protein level. shRNA interference was used to downregulate MORC2 in paclitaxel-resistant cells, and the actions of MORC2 reduction on paclitaxel resistance, cell cycle distribution, the uptake of glucose, and lactate production were investigated. As shown in A2780/Taxol and SKOV3/Taxol cells, MORC2 was highly expressed, and MORC2 protein expression was related to the time and concentration of paclitaxel. MORC2 downregulation made paclitaxel-resistant ovarian cancer cells more sensitive to paclitaxel, which indicated that more cells were arrested at the G0/G1 phase, suppressed glucose metabolism-related indicators (GLUT4, LDHA, and HK2). MORC2 knockdown reduced tumor growth and glycolysis, and enhanced paclitaxel sensitivity in vivo. These findings suggest MORC2 is upregulated in paclitaxel-resistant ovarian cancer cells, and that downregulation of MORC2 may reduce glycolysis and improve sensitivity to paclitaxel should be further examined.

This study aimed to investigate the regulatory role of chordin-like 2 (CHRDL2) in the Th17/Treg balance and its impact on the progression of osteoarthritis (OA). We evaluated the levels of CHRDL2 and Th17/Treg-related cytokines, and the proportions of Th17 and Treg cells in peripheral blood from both healthy subjects and OA patients. An OA mouse model was established by destabilization of the medial meniscus (DMM) surgery, and lentivirus-mediated overexpression and knockdown of CHRDL2 were conducted. The clinical and pathological manifestations of the mice were assessed, and knee joint cartilage damage was evaluated using histological staining. Additionally, we examined the levels of Treg/Th17-related inflammatory factors and transcription factors in peripheral blood, as well as the Treg/Th17 ratio. In both OA patients and mice, CHRDL2 expression was downregulated, with a significant increase in Th17 cell proportion and IL-17 levels, while Treg cell proportion and IL-10 levels were significantly decreased. Overexpression of CHRDL2 significantly improved the clinical and pathological manifestations in OA mice, corrected the Th17/Treg imbalance, reduced IL-17 and RORγt levels, and increased IL-10 and Foxp3 levels. However, knockdown of CHRDL2 results in the opposite effect. This study demonstrates that CHRDL2 can suppress OA progression by regulating the Th17/Treg balance and may serve as a key therapeutic target for alleviating immune dysregulation in OA.

Prostate cancer (PC) continues to represent a significant contributor to male cancer mortality worldwide, necessitating the discovery of innovative diagnostic indicators and molecular targets. Our investigation utilized computational biology approaches combining multi-omics analysis with machine intelligence to elucidate the role of palmitoylation-related genes (PRGs) in PC pathogenesis and prognosis. By harmonizing transcriptomic datasets from TCGA and GEO repositories, we identified dysregulated PRGs and stratified PC into two molecularly distinct subtypes via unsupervised clustering. These subtypes exhibited divergent clinical outcomes, immune microenvironment heterogeneity (e.g., Dendritic cells, CD8⁺ T cells, and Macrophages infiltration), and distinct drug sensitivity profiles. Single-cell RNA sequencing further localized key PRGs-ZDHHC2, ZDHHC5, ZDHHC15, ZDHHC9, and LYPLA1-within tumor cell populations, linking their expression to immune evasion and metabolic reprogramming. A robust diagnostic model, integrating 101 machine learning algorithms, demonstrated high predictive accuracy for survival and immunotherapy response. Functional validation in DU145 cells confirmed that modulating these PRGs significantly suppressed proliferation and colony formation, highlighting their pathobiological relevance. Collectively, this multidimensional analysis delineates a comprehensive framework for understanding palmitoylation-driven oncogenesis and establishes a precision medicine toolkit for risk stratification and treatment optimization in PC.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00842-3.

This study delves into the osteogenic properties of Rhizoma Drynariae, a traditional Chinese medicinal herb known for facilitating bone repair. Rhizoma Drynariae-derived exosomes were isolated and investigated to understand their impact on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). The exosomes significantly enhanced differentiation by elevating the expression of osteogenic genes such as RUNX2, OPN, and OCN, and promoting calcium nodule formation. Network pharmacology and bioinformatics analysis identified 86 pivotal genes, pinpointing the critical role of the TNF signaling pathway and inflammatory response in this enhancement. Functional validation showed that TNF signaling inhibition reduced osteogenic gene expression, mineralization, and ALP activity, confirming its essential role. These findings reveal the intricate molecular interactions underpinning the effects of Rhizoma Drynariae exosomes, providing a foundation for developing innovative therapeutic strategies to treat bone disorders and regenerate damaged bone tissue effectively. Graphical Abstract. Experimental Validation and Potential Molecular Mechanisms of Rhizoma Drynariae -Exosomes in Promoting Osteogenic Differentiation of BMSCs.

Graphical abstract: Experimental Validation and Potential Molecular Mechanisms of Rhizoma Drynariae -Exosomes in Promoting Osteogenic Differentiation of BMSCs.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00860-1.

Background: The high glucose (HG) levels in the body of a patient with gestational diabetes mellitus (GDM) constantly stimulate the trophoblast cells, leading to impaired development of the placenta thus affecting the development of the embryo. Previous analyses have indicated that the expression of microtubule-associated protein 6 domain containing 1 (MAP6D1) was lower in placental tissues of healthy pregnant women than that in GDM patients, but its specific effects and potential molecular mechanisms have not been elucidated.

Methods: HTR-8/SVneo cells were stimulated using HG to mimic the stimulation of trophoblast cells by the high glucose environment in vivo, and MAP6D1 expression was inhibited by transfection of MAP6D1 small interfering RNA (siRNA). Subsequently, cell proliferation and apoptosis levels were detected by methylthiazolyldiphenyl-tetrazolium bromide (MTT) and flow cytometry (FCM). Cell metastatic capacity was assessed by cell scratch assay and Transwell analysis. Enzyme linked immunosorbent assay (ELISA) was used to analyze the level of cellular inflammatory response. In addition, the degree of cellular oxidative stress was assessed by malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) levels as well as ROS fluorescence intensity. Western blot assay was used to analyze p38 MAPK pathway changes.

Results: HG induced elevated MAP6D1 expression in HTR-8/SVneo cells, inhibited cell viability and metastatic ability, and promoted apoptosis, while HG stimulation was able to promote cellular inflammatory responses and oxidative stress levels. In addition, HG treatment promoted the level of p38 MAPK protein phosphorylation in HTR-8/SVneo cells. However, inhibition of MAP6D1 expression was able to reverse the above results.

Conclusion: Inhibition of MAP6D1 expression ameliorated trophoblast cell injury induced by HG stimulation, providing a potential novel target for the diagnosis and treatment of GDM.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00866-9.

Natural killer (NK) cells are being developed as therapeutic agents targeting hematological malignancies and solid tumors. However, the lack of an optimal cryopreservation medium poses a significant challenge, as cryopreservation often reduces NK cell recovery, viability, and function, hindering their use in cellular therapies. This study investigated the effects of varying concentrations of Dimethyl sulfoxide (Me₂SO), Proline, Trehalose, and Dextran 40 commonly used in cell cryopreservation on NK cell recovery, viability, and cytotoxic activity. Additionally, we conducted a screening of 19 cryoprotective agents (CPAs) to enhance NK cell cytotoxic activity after freeze-thawing. We found that reducing Me₂SO concentration significantly decreased NK cell cytotoxic activity, and the combination of Proline, Trehalose, and Dextran 40 was insufficient to prevent this decline. A negative interaction effect between Trehalose and Dextran 40 on NK cell cytotoxic activity was also observed. Screening results identified Betaine, Glycine, Polyvinylpyrrolidone (PVP), α-tocopherol, Poloxamer 188, and Creatine as effective in enhancing NK cell cytotoxic activity after freeze-thawing. These findings provide new insights into the interaction effects of CPAs on NK cell cytotoxic activity and contribute to improving NK cell quality in pharmaceutical manufacturing.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00865-w.