Cytotechnology最新文献

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.

Gastric cancer (GC) remains among the leading causes of cancer-related mortality worldwide. Chronic alcohol consumption is a known risk factor for GC, but direct experimental evidence supporting alcohol as an independent oncogenic driver remains insufficient. In this study, we established a subcutaneous xenograft tumor model in nude mice by injecting MKN-45 cells and utilizing phorbol-12-myristate-13-acetate (PMA) to differentiate THP-1 cells into M0 macrophages. MKN-45 cells were then cultured with conditioned medium from alcohol-treated macrophages to evaluate malignant behaviors. Our findings demonstrate that alcohol consumption accelerated tumor growth in nude mice and increased the expression of Ki67 and epithelial‒mesenchymal transition (EMT) markers (N-cadherin and vimentin) but decreased the expression of E-cadherin. Furthermore, alcohol suppressed the expression of M1 macrophage polarization markers (iNOS, CD16, and CD86) and increased the expression of M2 markers (Arg1, CD163, and CD206), thereby promoting GC cell proliferation and migration. Notably, the expression of miR-29c-3p was downregulated in the blood of patients with GC who consumed alcohol and in the tumor tissues of alcohol-exposed mice. The overexpression of miR-29c-3p in macrophages reversed alcohol-induced effects by promoting M1 polarization and inhibiting M2 polarization. Mechanistically, alcohol suppresses miR-29c-3p expression in macrophages, leading to increased COL1A1 expression, which drives M2 polarization and accelerates GC progression.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00862-z.

CircRNAs are associated with the progression of various tumors. This study aims to explore the regulation of circUSP36 on cervical cancer progression and the mechanism behind. Human cervical cancer cell lines HeLa and SiHa were cultured for RT-qPCR detection of circUSP36, miR-520d-5p, and FBXO5 mRNA and western blotting detection of FBXO5 protein. After transfection, HeLa and SiHa cells were subjected to CCK-8, transwell, scratch, and flow cytometry assays to measure their proliferation, invasion, migration, and apoptosis. Luciferase reporters were used to detect the binding of miR-520d-5p to circUSP36 and FBXO5. CircUSP36 and FBXO5 were highly expressed in HeLa and SiHa while miR-520d-5p was expressed at low levels. CircUSP36 or FBXO5 knockdown or miR-520d-5p overexpression reduced proliferation, invasion, and migration and increased apoptosis in cervical cancer cell lines. CircUSP36 targeted miR-520d-5p and inhibited its expression. miR-520d-5p negatively regulated FBXO5 expression by binding to FBXO5. miR-520d-5p inhibition or FBXO5 overexpression reversed the tumor-suppressive effect of circUSP36 knockdown. CircUSP36 promotes FBXO5 expression by targeting and inhibiting miR-520d-5p, thereby driving cervical cancer cell proliferation, invasion, and migration.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00852-1.

Melanoma is an aggressive and highly metastatic skin cancer characterized by high mortality and resistance to conventional chemotherapy. Aberrant activation of the PI3K and MAPK signaling pathways in melanoma cells enhances survival and drives tumor progression. Targeting autophagy has emerged as a novel therapeutic strategy to induce cell death in melanoma. In this study, we evaluated IIIM-321, a synthetically derived lipidated analog of liphagal, which effectively downregulated both MAPK and PI3K signaling pathways, leading to a time- and dose-dependent reduction in melanoma cell proliferation. IIIM-321 induced significant pro-apoptotic effects in B16F10 cells, accompanied by autophagy inhibition, reactive oxygen species (ROS) accumulation, modulation of antioxidant defenses, and mitochondrial membrane potential (ΔΨM) disruption. The compound induced G0/G1 cell cycle arrest through upregulation of p21, downregulation of Cyclin D1, and its degradation via phosphorylated GSK3β, ultimately triggering apoptosis. Apoptotic induction was further confirmed by annexin-V/PI staining, cytochrome c release, caspase activation, and PARP cleavage. IIIM-321-mediated autophagy inhibition was evidenced by increased LC3-II, decreased Beclin1, and p62/SQSTM1 accumulation. These findings were supported by acridine orange staining and immunocytochemistry for Beclin1 and p62. Additionally, IIIM-321 suppressed both basal and α-MSH-stimulated melanogenesis in B16F10 cells, along with reduced expression of Tyrosinase, Trp-1, and Trp-2. Collectively, our findings establish IIIM-321 as a promising anti-melanoma agent with therapeutic potential for future clinical development.

Graphical abstract:

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00834-3.

Circular RNAs (circRNAs) represent a distinct category of endogenous non-coding RNAs, distinguished by their covalently closed-loop structure, which imparts remarkable stability and tissue-specific expression. Historically perceived as less prevalent than linear RNAs, recent findings indicate that circRNAs are integral to various biological processes. A well-documented function of circRNAs is their capacity to serve as microRNA (miRNA) sponges; however, recent investigations have revealed their participation in protein interactions and the potential for translation into functional peptides, thereby broadening their functional scope. Researchers have found that circRNAs play a big role in controlling the growth of adipose tissue in cattle, but we still don't fully understand the complex networks and downstream pathways they affect. This review aims to consolidate existing knowledge and provide new insights into the mechanistic roles of circRNAs in bovine adipogenesis, highlighting their effects on preadipocyte proliferation, differentiation, and lipid metabolism. To address this gap, a systematic literature search was performed utilizing databases such as ScienceDirect, PubMed, Google Scholar, and Web of Science to gather recent advancements in circRNA research pertinent to bovine adipogenesis. Key areas of focus include circRNA biogenesis and function, circRNA-mediated regulation of adipogenesis, molecular signaling pathways, epigenetic modifications, and the functional diversity of adipogenic regulation. This comprehensive review synthesizes current findings, identifies knowledge gaps, and delineates future research trajectories, particularly concerning the regulatory networks and downstream effectors associated with circRNAs. By elucidating the molecular mechanisms that underlie circRNA function in bovine adipogenesis, this review contributes to the fields of molecular biology, agricultural biotechnology, and veterinary science. Additionally, it highlights the potential of circRNAs as innovative biomarkers and therapeutic targets for enhancing cattle adipose tissue composition and meat quality, thereby establishing a foundation for future translational research in livestock genetics and metabolic regulation.