Cytotechnology最新文献

Resveratrol-rich extracts from Vitis vinifera cane have gained recognition as natural compounds with significant anticancer potential. This study examined the impact of this extract on A549 lung cancer cells using both 2D monolayer culture and a more physiologically relevant 3D co-culture model incorporating peripheral blood mononuclear cells (PBMCs). In monolayer culture, at 40 µg/mL we evaluated the extract's impact on cell viability, colony formation ability, morphological changes, and oxidative stress markers. A 3D co-culture model was employed to examine the effects of the extract on matrix metalloproteinase ( MMP-2 and MMP-9) activity, c-Jun N-terminal kinase (JNK) signaling pathway, and nitric oxide (NO) production. Our results demonstrated that the extract significantly reduced A549 cell viability (IC50 39.11 ± 0.32 µg/mL) and colony formation while inducing morphological changes and cellular redox imbalance markers malondialdehyde (MDA) (p ≤ 0.001), protein carbonyl (p ≤ 0.0001), Advanced Oxidation Protein Products (AOPP) (p ≤ 0.01), nitric oxide (NO) (p ≤ 0.0001), Superoxide Dismutase (SOD) activity (p ≤ 0.01), and glutathione (GSH) level (p ≤ 0.01). and in monolayer cultures. In the 3D co-culture system, treatment with the extract significantly decreased the growth rate of the 3D structures (P ≤ 0.1), reduced MMP-2 and MMP-9 activities (P ≤ 0.1), and JNK phosphorylation, with an overproduction of nitric oxide (P ≤ 0.001). These findings reveal novel mechanisms by which resveratrol-rich extracts from Vitis vinifera cane may exert anticancer effects by modulating the JNK/MMPs/NO axis in lung cancer cells A549. This extract could represent a promising therapeutic alternative for lung cancer treatment.

Endometrial cancer (EC) is one of the most pervasive gynecological malignancies, with its incidence increasing annually. Although progress has been made, the underlying molecular mechanisms driving EC progression remain poorly understood. N⁶-methyladenosine (m⁶A) RNA modification is instrumental in tumor biology, but the function of the m⁶A "writer" RNA-binding motif protein 15 (RBM15) in EC remains unclear. This study investigates the role of RBM15 through m⁶A-dependent regulation of its downstream target. RBM15 expression was analyzed using the TNMplot database and validated by qRT-PCR in EC tissues and cell lines. Cell functional assays were conducted to assess the impact of RBM15 knockdown. Bioinformatics analytics and MeRIP-qPCR were used to identify m⁶A-modified downstream targets of RBM15. Furthermore, RNA stability assay, correlation analysis, and Western blotting were employed to explore the underlying regulatory mechanisms. Rescue experiments were done to confirm functional interactions. RBM15 was significantly overexpressed in EC, and its silencing impeded EC cell proliferation, migration, and invasion. PBK was identified as a direct downstream target, with RBM15-mediated m⁶A enrichment enhancing mRNA stability. PBK expression was positively correlated with RBM15 levels in patient samples. RBM15 silencing partially attenuated the PBK-driven promotive effects in EC cells. Mechanistically, RBM15 suppressed the p53 pathway by stabilizing PBK, resulting in decreased phospho-p53 protein levels. In summary, RBM15 promotes EC progression by enhancing m⁶A-dependent stabilization of PBK mRNA, which in turn suppresses p53 signaling, revealing a novel RBM15-PBK-p53 axis as a potential treatment target.

Ovarian cancer (OC) is a type of gynecological cancer, characterized by a high mortality rate. MicroRNAs (miRNAs) and cell autophagy are associated with the chemoresistance of tumors. The purpose of this study was to investigate the role of miR-675-5p in cisplatin (DDP) resistance in OC cells and its related mechanism. We collected OC tissues and paracancerous tissues. The OC cell line A2780 and the DDP-resistant cell line A2780/DDP were used for the study. RT-qPCR, Western blot and immunofluorescence were used to detect the expression of related genes and proteins. Cell proliferation and apoptosis were evaluated by CCK-8 assay and flow cytometry. Furthermore, the effect of miR-675-5p on the progression of OC was investigated in a xenograft model of nude mice. In this research, miR-675-5p expression was significantly increased in OC tissues and A2780/DDP cells. The suppression of miR-675-5p in A2780/DDP cells resulted in a significant reduction in DDP resistance. Further study revealed that miR-675-5p inhibited TSC2 expression in OC cells by binding to the TSC2 3'UTR, thereby activating the mTOR signaling pathway and promoting autophagy in A2780/DDP cells. Knocking down miR-675-5p suppressed DDP resistance in A2780/DDP cells through TSC2/mTOR axis, while overexpressing miR-675-5p suppressed DDP sensitivity in A2780 cells through TSC2. The tumor volume was decreased after injection of miR-675-5p-knockdown A2780/DDP cells in vivo. Knocking down miR-675-5p can inhibit the mTOR signaling pathway and autophagy through TSC2, thereby reducing the DDP resistance of OC cells. This may provide a potential therapeutic target for OC.

[This retracts the article DOI: 10.1007/s10616-013-9534-x.].

Bladder cancer (BLCA) is a prevalent malignancy of the urinary tract. Long noncoding RNAs (lncRNAs) exert significant effects on various human cancers by targeting microRNAs (miRs). This study, therefore, probed the action of the LINC00152/miR-103a-3p axis in epithelial-mesenchymal transition (EMT) and progression of BLCA. LINC00152, miR-103a-3p, and ADORA3 expression in clinical BLCA tissues were tested. LINC00152 was knocked down in BLCA cells alone or combined with miR-103a-3p inhibition or ADORA3 overexpression. The level of EMT- and PI3K/AKT pathway-related genes was assessed with Western blot. Cell proliferation, invasion and migration, and apoptosis were examined with CCK8 and EdU, Transwell, and flow cytometry assays, respectively. Binding of miR-103a-3p to LINC00152 and ADORA3 was validated through RIP and dual-luciferase reporter gene assays. The action of LINC00152 in BLCA progression was verified in vivo. LINC00152 and ADORA3 expression was abundant and miR-103a-3p expression was poor in BLCA tissues. LINC00152 knockdown or PI3K/AKT pathway inhibition repressed malignant behaviors and EMT of BLCA cells. Mechanistically, LINC00152 bound specifically to miR-103a-3p, which targeted ADORA3 and then activated the PI3K/AKT pathway. miR-103a-3p inhibition or ADORA3 overexpression nullified the suppressive impacts of LINC00152 knockdown on EMT and malignant behaviors of BLCA cells. Furthermore, LINC00152 knockdown decreased ADORA3 expression and suppressed tumor growth in vivo. LINC00152 knockdown upregulates miR-103a-3p to reduce ADORA3 expression, thus protecting against the malignant biological behaviors and EMT of BLCA cells.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00829-0.

Super-resolution fluorescence microscopy (SRM) has enabled visualization of nanoscale cellular structures, but systematic evaluation of resolution assessment methods across diverse biological structures and SRM modalities remains lacking. Here, we comparatively assessed three resolution metrics-Full Width at Half Maximum (FWHM), decorrelation analysis, and Fourier Ring Correlation (FRC)-across two SRM techniques (Super-resolution Radial Fluctuation, SRRF; Stimulated Emission Depletion, STED) using key subcellular structures: microtubules (filaments), mitochondria (membranes), and nuclear pore protein Nup98 (single particles) in HeLa/U2OS cells. Our results showed decorrelation analysis provided robust resolution estimates across all structures and modalities (confocal/SRRF/STED), exhibiting superior performance for dense nuclear pore complexes where FWHM failed due to overlapping point spread functions. FWHM was effective for discrete structures (microtubules/mitochondria) but unsuitable for densely packed features. NanoJ-SQUIRREL's integrated FRC analysis successfully quantified SRRF resolution for all structures, with resolution-scaled Pearson coefficient (RSP > 0.90) confirming minimal reconstruction artifacts. STED achieved significantly higher resolution than SRRF, but severe photobleaching prevented FRC application. Our study demonstrates decorrelation analysis offers universal robustness across structures and modalities, FWHM is suitable for discrete structures, and NanoJ-SQUIRREL provides standardized artifact validation and FRC-based resolution quantification for computational super-resolution. This study establishes a framework for context-appropriate resolution assessment in cell biology, emphasizing integration of structural features and technical principles.

Obesity has become a global public problem. Guizhi Fuling Decoction (GZFLD) is a traditional Chinese medicine decoction. This study aims to investigate the therapeutic effects of GZFLD on high-fat diet (HFD)-induced obesity in mice. The obese mouse model was induced by HFD. The effects of GZFLD on obesity were assessed by recording body weight (BW) and inguinal white adipose tissue (Ing-WAT) weight of mice. Histological changes in adipose and liver tissues were evaluated by hematoxylin-eosin staining. Enzyme-linked immunosorbent assay was exploited to detect the expression of blood lipid indexes and inflammatory factors. The expression of pyroptosis-related proteins was detected by western blot and immunohistochemistry. Transcriptome sequencing was executed to explore the mechanism of GZFLD on obese mice and screen the pyroptosis-related genes. Furthermore, the effect of Angiotensin-converting enzyme 2 (ACE2) on obese mice was verified by ACE2 inhibitor-DX600. In HFD-induced mice, with the increased dose of GZFLD, GZFLD significantly improved obesity, hepatic steatosis and dyslipidemia, with decreased BW, Ing-WAT weight, Ing-WAT/BW ratio, levels of triglycerides, cholesterol low-density lipoprotein, and increased high-density lipoprotein levels. Hematoxylin-eosin staining exhibited that GZFLD improved steatosis and adipocyte enlargement induced by HFD. Additionally, GZFLD suppressed the levels of lactate dehydrogenase, inflammation factors, and pyroptosis-related proteins. RNA-sequencing identified ACE2 as a key gene target. The ACE2 inhibitor DX600 attenuated the inhibitory effects of GZFLD on hepatic steatosis, dyslipidemia, inflammation, and pyroptosis in HFD-induced obese mice. Collectively, GZFLD had a protective effect on HFD-induced obesity by alleviating dyslipidemia, and inhibiting inflammation and pyroptosis, which was achieved by up-regulating ACE2.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00848-x.

Ischemic stroke (IS) is a leading cause of death and long-term disability worldwide; however, effective neuroprotective therapies remain limited. Glabridin, a flavonoid from licorice, exhibits antioxidant and anti-inflammatory properties, making it a candidate for stroke therapy. To evaluate the therapeutic potential and mechanisms of glabridin in IS, we combined computational and experimental approaches. Network pharmacology and molecular docking were used to predict potential targets and pathways of glabridin in ischemic stroke. Functional validation employed a transient middle cerebral artery occlusion/reperfusion (tMCAO/R) mouse model (behavioral testing, TTC, HE, TUNEL/Nissl staining, and biochemical assays) and an in vitro oxygen-glucose deprivation/reperfusion (OGD/R) model in HT22 neuronal cells (cell viability assays and Western blot analysis). We found that glabridin significantly reduced infarct volume and improved neurological and motor function in MCAO/R mice, decreased neuronal apoptosis, attenuated inflammatory and oxidative stress markers (TNF-α, MDA) and increased antioxidant activity (SOD). In HT22 cells glabridin reduced OGD/R-induced apoptosis and restored cell viability. Mechanistically, both molecular docking and biochemical assays indicated that glabridin activates the PI3K/Akt signaling pathway, which likely mediates its neuroprotective effects. In summary, our integrative study demonstrates that glabridin ameliorates ischemia/reperfusion injury through anti-apoptotic, anti-inflammatory and antioxidant actions associated with PI3K/Akt activation, supporting further preclinical development.

Pore membrane protein 121 (POM121) is highly expressed in non-small cell lung cancer (NSCLC) and is associated with tumor progression and metastasis. However, its role in NSCLC chemoresistance remains unclear. This study aimed to investigate the effect of POM121 on cisplatin (cis-diamminedichloroplatinum [CDDP]) sensitivity in NSCLC and to elucidate its potential molecular mechanisms. Real-time quantitative polymerase chain reaction and western blotting were used to detect the expression of POM121 in normal bronchial epithelial cells and various NSCLC cell lines (A549, H460, H1299, and H1975). POM121 was silenced in NSCLC cells and cisplatin-resistant A549/CDDP cells. Changes in cell viability, half-maximal inhibitory concentration values, and apoptosis rates were analyzed using Cell Counting Kit-8 and flow cytometry, respectively, and the expression of apoptosis-related proteins was detected using western blotting. POM121 expression was significantly upregulated in NSCLC cells. Silencing of POM121 markedly reduced the resistance of A549, H460, and A549/CDDP cells to cisplatin. Silencing of POM121 significantly induced apoptosis in A549, H460, and A549/CDDP cells and promoted apoptosis in cisplatin-induced A549/CDDP cells, suggesting that POM121 may mediate resistance by regulating apoptotic pathways. POM121 promotes cisplatin resistance in NSCLC; its inhibition could be a potential therapeutic strategy to enhance cisplatin efficacy.

Cytotoxic CD8⁺ T lymphocytes play a pivotal role in anti-tumor immunity by eliminating cancer cells. Sargentodoxa cuneata (Sar) has demonstrated anti-cancer potential. This study investigated the therapeutic potential of Sar in lung adenocarcinoma (LUAD) through its modulation of CD8⁺ T cell tumoricidal capacity. CD8⁺ T cells were isolated and co-cultured with treated HCC2935 and H1975 LUAD cell lines. The influences on LUAD cells were assessed by detecting cell viability and apoptosis. The impacts on CD8⁺ T cells were evaluated by measuring cell cytotoxic activity and IFN-γ and TNF-α secretion. The aminoacylase-1 (ACY1)/arginase-2 (ARG2) interaction was predicted by molecular docking and confirmed by GST pull-down and Co-IP assays. Animal xenograft experiments were used to analyze the therapeutic potential of Sar in vivo. ACY1 and ARG2 were upregulated and positively associated with PD-L1 expression in LUAD samples. Mechanistically, ACY1 physically interacted with ARG2 in LUAD cells. ACY1 inhibited apoptosis in LUAD cells and attenuated cytotoxic activity of CD8⁺ T lymphocytes via ARG2. Moreover, Sar induced LUAD cell apoptosis and enhanced CD8⁺ T cell cytotoxicity by downregulating ACY1 in vitro. Sar attenuated xenograft tumor development through ACY1 downregulation in vivo. Our study establishes that Sar emerges as a promising therapeutic agent in LUAD by enhancing CD8⁺ T tumoricidal capacity through targeting the ACY1/ARG2 co-regulatory axis.

Graphical abstract: Sargentodoxa cuneata (Sar) reduces ACY1 expression to disrupt the ACY1/ARG2 interaction and thus downregulates PD-L1, consequently inducing LUAD cell apoptosis and enhancing CD8⁺ T cell cytotoxicity.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00849-w.