Cytotechnology最新文献

Pancreatic ductal adenocarcinoma (PDAC) is a cancer with a very poor prognosis, which is bolstered by its resistance and pathogenesis. Quest for PDAC treatments has not left any stones unturned. A well-known naphthoquinone found in many foods, plumbagin (PLGN) demonstrated effectiveness in treating cancer, arthritis, dysmenorrhea, and injuries. We hypothesize therapeutic advantages of PLGN in PDAC and used web resources to assist us identify the targets of PLGN in order to test the idea. After that, in-vitro studies were carried out to comprehend PLGN impacts on two PDAC cell lines. Further, drug combination with PLGN and pre-sensitization by PLGN was also carried out. Findings from network pharmacology investigations revealed PLGN's targets in PDAC, indicating effect on PDAC survival by targeting crucial signalling. In-vitro tests demonstrated dose-dependent cytotoxicity of PLGN, with IC₅₀ values for Panc-1 and Mia-PaCa-2 cells after 48 h were 2.89 ± 0.42µM and 13.10 ± 0.76µM, respectively. Dox-PLGN and Gem-PLGN combination treatment had antagonistic effects, although doxorubicin's impact was amplified by PLGN pre-sensitization. The EMT transition and genes that cause resistance were the primary pathways impacted by PLGN therapy that enhance Dox effect. Analysis of gene expression and ROS production demonstrated that PLGN had a different effect on both cell lines. Overall, PLGN shows promise as a pre-sensitizing natural substance for PDAC treatment, promoting more research on its effectiveness, tolerability, and possible incorporation into current chemotherapy regimens.

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Supplementary information: The online version contains supplementary material available at 10.1007/s10616-026-00912-0.

Myocardial cell injury following myocardial infarction (MI) is closely associated with abnormal glycolysis metabolism. Shexiang-Tongxin Dropping Pill (STDP) exhibits potential cardioprotective effects, but its mechanism in regulating post-MI glycolysis remains poorly understood. This study aimed to elucidate the molecular mechanism by which STDP ameliorates MI-induced injury through integrin subunit beta 2 (ITGB2) methylation regulation, providing new therapeutic insights and potential targets. The in vivo MI model was generated by permanent ligation of the left anterior descending coronary artery in Sprague-Dawley rats, while the in vitro model was established through hypoxia/reoxygenation treatment in AC16 cardiomyocytes. Histopathological evaluation of myocardial injury and apoptosis was performed using hematoxylin and eosin staining and terminal deoxynucleotidyl transferase dUTP nick-end labeling assay, respectively. Cardiac functional impairment was quantified by measuring serum lactate dehydrogenase (LDH) and creatine kinase-myocardial band (CK-MB) levels. Western blot analysis was conducted to examine the expression of fibrosis-related proteins (collagen I, α-smooth muscle actin) and cardiac remodeling markers (atrial natriuretic peptide, brain natriuretic peptide). Cellular glycolytic metabolism was assessed by monitoring extracellular acidification rate and oxygen consumption rate using Seahorse metabolic analysis. Furthermore, methylation-specific PCR (MSP) combined with DNA immunoprecipitation was employed to investigate ITGB2 promoter methylation status. STDP significantly attenuated myocardial fibrosis, apoptosis, and cardiac dysfunction in MI rats, concurrently reducing serum LDH and CK-MB levels. It suppressed glycolytic activity and the expression of pyruvate kinase M2 (PKM2). Furthermore, STDP upregulated ITGB2 promoter methylation and decreased its expression. These protective effects were reversed by the methylation inhibitor 5-aza-2'-deoxycytidine. STDP promotes ITGB2 promoter methylation, thereby reducing its expression. This process is associated with the restoration of abnormal glycolysis and downregulation of PKM2, collectively contributing to the amelioration of myocardial injury post-MI.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-026-00911-1.

In the tumor microenvironment, hypoxia and stromal interactions contribute to enhanced malignant behavior in cancer cells. This study aimed to assess whether pancreatic cancer cells with higher malignancy display stronger responses to hypoxia and stromal cells than their less malignant parental cells, and evaluated the underlying mechanisms, focusing on lysophosphatidic acid (LPA) receptor signaling linked to the acquisition of malignant traits. Highly invasive PANC-M10 cells, derived from the parental pancreatic cancer PANC-1 cells, were cultured at 1% O₂ to mimic hypoxic conditions and co-cultured with lymphatic endothelial SVEC4-10 cells. Exposure to 1% O₂ increased LPAR2 and LPAR3 expression in PANC-M10 cells. Although cell proliferation in response to LPA treatment in 1% O₂ culture also increased in PANC-1 cells, the increase was more pronounced in PANC-M10 cells. PANC-M10 cells displayed markedly elevated invasive activity in 1% O₂ compared with PANC-1 cells. This hypoxia-induced invasion was reduced by AM966 (LPA₁ antagonist) and GRI-977,143 (LPA₂ agonist), while (2 S)-OMPT (LPA₃ agonist) further enhanced invasive capacity, indicating distinct receptor-dependent functions. Co-culture with SVEC4-10 cells at 1% O₂ amplified the invasive behavior of PANC-M10 cells beyond that observed under monoculture. In addition, the supernatant collected from PANC-M10 cells maintained at 1% O₂ more effectively stimulated SVEC4-10 tube formation than the supernatant from PANC-1 cells. These findings demonstrate that highly invasive pancreatic cancer cells undergo hypoxia-driven crosstalk with lymphatic endothelial cells, promoting tumor progression through LPA receptor-mediated signaling pathways.

Colorectal cancer (CRC) remains a major cause of cancer-related mortality worldwide, largely due to therapeutic resistance and the complexity of the tumor microenvironment. The extracellular matrix (ECM) critically regulates tumor progression by balancing matrix metalloproteinases (MMPs) and their inhibitors (TIMPs). Theranekron^® D6, an extract of Tarantula cubensis with known anti-inflammatory and wound-healing effects, has shown anticancer potential. This study investigated its molecular impact on ECM remodeling and apoptosis in human CRC. Human colorectal adenocarcinoma (Caco-2) and non-tumoral embryonic kidney (HEK293) cells were treated with Theranekron^® D6 (1-1000 µL/mL). Cell viability was evaluated using the MTT assay, revealing a dose- and time-dependent cytotoxic response with an IC₅₀ of 400 µL/mL at 48 h. Gene expression of 62 target genes spanning ECM remodeling, apoptosis, and key signaling axes (PI3K/AKT, NF-κB, and SMAD/STAT) was analyzed by quantitative polymerase chain reaction (qPCR), and protein levels of TIMP1, MMP2, MMP9, MMP14 and MMP28 were quantified by enzyme-linked immunosorbent assay (ELISA). Pathway interactions were assessed via STRING-based enrichment and clustering analyses. Theranekron^® markedly upregulated pyruvate dehydrogenase kinase 1 (PDK1) by 23-fold, with concurrent upregulation of PTEN suggesting a negative feedback mechanism that may limit net pro-survival PI3K/AKT output. This regulation restored a TIMP1-dominant ECM profile and reduced MMP2 and MMP28 protein levels. NF-κB pathway components were broadly downregulated at the transcriptional level. Concurrently, Theranekron^® induced transcriptional changes associated with mitochondrial and SMAD-dependent apoptotic pathways. Theranekron^® acts as a cell-context-specific modulator, simultaneously regulating PI3K/AKT/PDK1, NF-κB and SMAD pathways to restore ECM integrity and modulate apoptotic signaling pathways in Caco-2 CRC cells, suggesting its potential as a multi-target experimental therapeutic candidate in in vitro CRC models.

Diabetic nephropathy (DN) is one of the most serious and common diabetes-associated complications. Licochalcone B is a chalcone that has been found to possess anti-diabetic activity. However, protective potential of licochalcone B in DN remains unclear. In this study, we investigated the protective effect of licochalcone B against DN both in vitro and in vivo. The results showed that licochalcone B protected HK-2 cells from high glucose (HG)-induced oxidative stress and ferroptosis. Further molecular docking and molecular dynamics simulation revealed the stable binding abilities of licochalcone B and activating transcription factor 3 (ATF3). We found that licochalcone B negatively regulated ATF3 expression. Licochalcone B also inhibited the HG-induced ATF3 expression in HK-2 cells. ATF3 overexpression reversed the inhibitory effects of licochalcone B on oxidative stress and ferroptosis in HG-induced HK-2 cells. Moreover, ATF3 overexpression downregulated the expression levels of solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4), which are two critical regulators of ferroptosis. In vivo assays also proved that licochalcone B administration significantly improved histopathological changes in kidney tissues from HFD/STZ mice. Overall, our findings suggested that licochalcone B exerted protective effect against DN through ameliorating oxidative stress and ferroptosis via regulating ATF3/SLC7A11/GPX4 axis. These results highlighted the therapeutic potential of licochalcone B in preventing and treating DN.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-026-00910-2.

[This corrects the article DOI: 10.1007/s10616-025-00847-y.].

Benign prostatic hyperplasia (BPH) is an increasingly prevalent pathology in adult men. We investigated the mechanism of suppressor of cytokine signaling 6 (SOCS6)/signal transducer and activator of transcription (STAT) 6 pathway regulating the proliferation of prostate epithelial cells under inflammation. In vitro cultured prostate epithelial cells (RWPE-1) were treated with 1 µg/mL lipopolysaccharide for 48 h to simulate an in vitro inflammatory environment and then treated with OE-SOCS6, si-SOCS6, si-STAT6 and their negative control, and a STAT6 inhibitor (AS1517499). SOCS6 expression and cell proliferation were assessed by RT-qPCR, Western blot, CCK-8 and EDU assays. The expression of proliferative proteins Ki-67, MCM7 and PCNA, along with cell cycle were determined by Western blot and flow cytometry. SOCS6 suppressed the proliferation of prostate epithelial cells under inflammation, while SOCS6 knockdown caused the opposite result. SOCS6 downregulation increased STAT6 phosphorylation level, stimulated STAT6 signaling pathway, and promoted progression of prostate epithelial cells under inflammation. Meanwhile, STAT6 knockdown decreased the STAT6 phosphorylation level, impeded the cell cycle progression of prostate epithelial cells under inflammation, and attenuated the cell viability, weakened the cell fluorescence intensity, and notably reduced the expression of proliferative proteins, indicating that STAT6 knockdown suppressed the prostate epithelial cell proliferation. Subsequently, STAT6 inactivation partially reversed the regulatory role of SOCS6 in cell cycle and prostate epithelial cell proliferation under inflammation. SOCS6 repressed prostate epithelial cell proliferation under inflammation, and knockdown of SOCS6 facilitated cell cycle progression by activating STAT6 signaling, thereby accelerating prostate epithelial cell proliferation.

Renal ischemia/reperfusion (I/R) injury is a serious medical condition that causes acute kidney dysfunction and is associated with a high mortality rate. Ferroptosis is a form of nonapoptotic cell death marked by excessive lipid peroxidation. This study aimed to investigate the effect of miR-34c-5p/HSPA1B axis on ferroptosis in in hypoxia/reoxygenation (H/R)-stimulated human renal proximal tubular cells (HK-2) and I/R rat models. Erastin was employed to trigger ferroptosis, while Ferrostatin-1 was applied to suppress it. The viability of HK-2 cells was evaluated using cell counting kit-8 assays, whereas apoptosis was detected via TUNEL assays. The levels of mitochondrial superoxide, lipid reactive oxygen species (ROS), and iron concentration were measured as indicators of ferroptosis. Luciferase reporter assays and RNA pulldown assays were utilized to measure the binding relation between miR-34c-5p and HSPA1B. Hematoxylin-eosin staining of rat kidney tissues was performed to evaluate renal pathological changes. H/R treatment induced intracellular ferroptosis, and the inhibition of ferroptosis via Ferrostatin-1 attenuated H/R-induced cell injury. HSPA1B was upregulated in the established cell model and promoted intracellular ferroptosis. MiR-34c-5p targeted HSPA1B in HK-2 cells, leading to a reduction in intracellular ROS production, Fe²⁺ level, and the rate of apoptosis. Importantly, miR-34c-5p mitigated renal histological damage mediated by I/R injury through the suppression of ferroptosis. In conclusion, miR-34c-5p inhibits ferroptosis in renal I/R injury by targeting HSPA1B.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-026-00933-9.

This study aimed to elucidate the active components and therapeutic mechanism of BuZhong Yulin decoction (BZYLD) against recurrent urinary tract infections (rUTI) by integrating UPLC-Q-TOF-MS analysis with network pharmacology. The chemical components of BZYLD were characterized in vitro and in vivo using UPLC-Q-TOF-MS. Potential Targets of the absorbed serum components were predicted using the Swiss Target Prediction database, while rUTI-related targets were retrieved from GeneCards and DisGeNET. A protein-protein interaction (PPI) network was constructed using the STRING database and visualized with Cytoscape 3.6.0. Core targets were identified and subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Finally, the key findings were experimentally validated in a cellular model. We identified 118 components in BZYLD formulation and 21 prototype components in serum. Network pharmacology analysis revealed that key active components included 7,4'-hydroxyflavone(7,4'-DHF), ferulic acid (FA), and 7-methoxycoumarin(7-MC), with IL-6 and TNF emerging as pivotal targets. KEGG pathway analysis highlighted significant enrichment in the PI3K-Akt and MAPK signaling pathways. Consistently, in the cellular assays, BZYLD and its key components (7,4'-DHF, FA, and 7-MC) significantly inhibited the release of IL-6 and TNF-α in infected RAW macrophages. Furthermore, they regulated the expression of proteins in the implicated the expression of key proteins in the implicated pathways, namely PI3K, MEK, and P38. Our integrated approach identified the active components of BZYLD and demonstrated that its anti-rUTI effects are likely mediated through the suppression of key inflammatory mediators via modulation of the PI3K-Akt and MAPK signaling pathways. These findings provide a robust pharmacological foundation for the clinical application of BZYLD.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-026-00928-6.

A protein called cytolysin A or ClyA, encoded by certain bacteria species, can cause cytotoxicity. Although the ClyA protein is not typically expressed at detectable levels in most E. coli strains, here it was successfully overproduced and purified by cloning the structural gene into an hns mutant strain. The cytotoxicity of the purified cytolysin was assessed on two MCF-7 cancer cell lines and HDF normal cell line using the MTT assay. Flow cytometry was employed to examine the cytolysin's ability to induce apoptosis in cancer cells. In addition, a Western blot analysis was carried out to evaluate the expression levels of P53, Bcl2, and Bax proteins. The results revealed that cytolysin exhibited dose-dependent and time-dependent toxicity towards cancer cells, while showing minimal toxicity against normal cells, indicating its selective action against cancer cells. Cytolysin had an IC50 value of 3.29 µg/ml against MCF-7 cancer cells and 12.6 µg/ml against HDF normal cells. Flow cytometry results further demonstrated that cytolysin induced apoptosis in cancer cells, evidenced by increased expression of p53 and BCL2, as well as decreased in Bax, in gene and protein levels. These findings underscore the potential of cytolysin as a targeted therapy for cancer, highlighting its selective cytotoxic effect on cancer cells.