Cytotechnology最新文献

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.

Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder. This study aims to investigate the expression of miR-103a-3p in PCOS and its potential molecular mechanisms. RT-qPCR was used to detect the levels of miR-103a-3p in the serum of PCOS patients, and to analyze its correlation with clinical indicators. The diagnostic value of miR-103a-3p for PCOS was evaluated through the ROC curve. Subsequently, the viability of KGN cells, inflammatory factors, and oxidative stress indicators were detected using CCK-8, ELISA, and oxidative stress detection kits, respectively. Finally, the targeting relationship between miR-103a-3p and PTEN was validated using dual luciferase reporter and RIP assays. The expression of miR-103a-3p in the serum of PCOS patients was significantly lower than that in the control group. miR-103a-3p demonstrates high sensitivity and specificity for the diagnosis of PCOS (AUC = 0.881). The expression level of miR-103a-3p is correlated with clinical indicators. In KGN cells, overexpression of miR-103a-3p significantly enhances cell viability, inhibits the secretion of inflammatory factors, and reduces oxidative stress levels, whereas inhibition of miR-103a-3p exhibits the opposite effects. The dual luciferase reporter assay and RIP experiment confirmed the direct interaction between miR-103a-3p and PTEN. In PCOS patients, miR-103a-3p is expressed at low levels and is closely associated with hormonal and metabolic indicators, potentially serving as an early diagnostic marker for PCOS. In vitro studies have found that miR-103a-3p may inhibit the progression of PCOS inflammation by targeting PTEN.

Our study focuses mainly on identifying the hepatoprotective activity of Naregamia alata ethyl acetate (NAEA) extract on Wistar rats. In addition, tangeretin was isolated, characterized and studied for in vivo and in silico approaches. D-GalN-induced hepatotoxicity rats were treated with 200 and 400 mg/kg of NAEA and its antioxidant and hepatoprotective activity was determined. The active constituent in the extract was isolated and spectral characterization was carried out. Cytoprotective, antioxidant and hepatoprotective activity of tangeretin was analyzed using MTT assay, DCFA-ROS assay and D-GalN induced toxicity in HepG2 cells. The anti-inflammatory activity of tangeretin in D-GalN treated cells was assessed by measuring the level of IL-6 and TNF-α via qRT-PCR. Molecular docking of tangeretin with the TACE enzyme was performed using AutoDock tools. Acute toxicity study in rats shows that NAEA exhibits no toxicity up to 4000 mg/kg. Hepatoprotective activity of the extract was confirmed by histopathological analysis and liver enzymes in d-galactosamine-induced hepatotoxicity rats treated with 200 and 400 mg/kg of NAEA. Spectral characterization reveals that the active constituent is tangeretin and MTT assay reveals IC₅₀ of 44.14 µM. 21.25 and 42.5 µM of tangeretin show antioxidant activity in DCFH-DA staining. The level of IL-6 and TNF-α were downregulated by tangeretin in HepG2 cells pretreated with D-Galactosamine. Molecular docking studies show that tangeretin binds to TACE with the binding energy of -9.13 kcal/mol and exhibits a low inhibition constant of 204.12 nM. Our findings show that the antioxidant, anti-inflammatory and hepatoprotective activity of Naregemia alata is due to the presence of tangeretin.

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Supplementary information: The online version contains supplementary material available at 10.1007/s10616-026-00902-2.

Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related mortality. Circulating tumor cells (CTCs) are widely recognized as the origin of hematogenous tumor metastasis. Cortactin (CTTN) plays a critical role in epithelial-mesenchymal transition, which is a critical step for malignant progression of tumor. This study aims to clarify the correlation between CTCs and recurrence in HCC. A total of 80 clinical patients were participated in this study and received 50 qualified peripheral blood samples. Patient clinical basic information was collected, CTC cells are isolated and counted at baseline to analyze the sensitivity and correlation of CTC counts as a patient recurrence biomarker. In-situ cancer mice of HCC were used to observe the effects of CTTN overexpression and knockdown on CTCs. The sensitivity and specificity of CTCs detection (>4.5/5 mL) were 90% and 82.5% (AUC = 0.871). Patients were divided into the CTC-Low group (n = 34) and CTC-High group (n = 16) at CTCs counts >4.5/5 mL. CTC >4.5/5 mL was associated with recurrence with a chi-square statistic (χ²) of 19.324 and p <0.001. The effect size, quantified by Cramer's V, was 0.622. CTTN is expressed on CTCs of HCC patients and CTTN overexpression promoted in-situ cancer growth and metastasis, increasing CTC numbers in HCC mice, while CTTN knockdown has the opposite effects. This study provides a research basis for the identification of clinical biomarker of CTTN for predicting the prognosis of patients with HCC after surgery and offers potential targets for the blockade therapy of CTCs.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00879-4.

Marine-derived functional ingredients are gaining attention for their therapeutic potential in skin health and inflammation. The wound healing and anti-inflammatory properties of Holothuria atra, an edible sea cucumber, remain underexplored. This study aimed to evaluate the in vitro wound healing activity of H. atra methanol extract and the in silico anti-tumor necrosis factor-α (TNF-α) of its metabolites. Cytotoxicity on human keratinocyte (HaCaT) cells was assessed, followed by scratch assays under normal and TNF-α-induced conditions. The components of the extract were identified based on their molecular masses, and key metabolites were docked against TNF-α binding sites. The extract showed 83.8 ± 2.5% cell viability at 100 ng/mL, which was used as the maximum concentration. In the scratch assay, wound closure increased with dose, reaching 53.2 ± 8.8% after 48 h at 100 ng/mL, compared to 18.4 ± 6.9% in the control (p < 0.0001). With TNF-α (10 ng/mL), cell migration dropped (1.0 ± 0.2% after 48 h); however, 100 ng/mL extract restored closure to 53.2 ± 15.1% (p < 0.001) after 48 h. GC-MS revealed major fatty acids, including arachidonic (17.5%) and palmitic acid (15.2%), while LC-MS identified ten saponins, six verified by standards. Docking showed metabolite binding to TNF-α (-9.29 to - 7.47 kcal/mol), particularly at Tyr119. In conclusion, H. atra methanol extract enhanced keratinocyte wound healing in vitro, and its metabolites showed TNF-α binding ability based on the molecular docking study. However, further in vivo validation is warranted.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00880-x.

Knee osteoarthritis (OA) is a prevalent degenerative joint disease and a leading cause of disability worldwide. While traditionally considered a mechanical disorder, accumulating evidence highlights the critical role of dysregulated immune responses in OA pathogenesis. Despite this, specific immune-related biomarkers for early diagnosis remain poorly defined. This study aims to systematically identify and validate immune-related molecular signatures involved in knee OA, providing potential diagnostic biomarkers and therapeutic targets. We applied integrated bioinformatics and machine-learning approaches to identify immune-related biomarkers specific to OA and validated their expression through in vitro experiments. Two immune-related hub genes, BCL6 and HMGB2, were found to be significantly downregulated in OA cartilage. Both demonstrated strong diagnostic performance and were consistently validated across external datasets and in vitro assays. This study identifies BCL6 and HMGB2 as robust immune-related biomarkers in knee OA, providing new insights into disease mechanisms and potential avenues for early diagnosis and therapeutic intervention.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00876-7.

This study was to investigate the therapeutic effect of Panax notoginseng-Bletilla striata (PN-BS) in reflux esophagitis (RE) and its molecular mechanism. Using the '4.2 mm pyloric clamp + 2/3 fundoplication' method, a rat model of RE was developed. RE cell model was established by exposing HET-1 A (esophageal epithelial cells) to bile salt. Esophageal mucosal injury was observed by HE staining, and epithelial barrier dysfunction was assessed using Toluidine blue staining. HET-1 A cell viability was measured by CCK-8. Inflammatory factors in tissues and cells were detected by enzyme-linked immunosorbent assay. Claudin-4, Claudin-5, NLRP3, cleaved-caspase-1, p-p38 MAPK, and p38 MAPK protein levels were detected by Western blot. PN-BS attenuated esophageal mucosal injury and inflammation and improved esophageal barrier dysfunction in RE rats. Panax notoginseng saponins (PNS, the main active ingredient of PN) and Bletilla striata polysaccharides (BSP, the main active ingredient of BS) attenuated acid and bile salt-induced esophageal barrier dysfunction. PNS and BSP inhibited NLRP3 inflammasomes and p38 MAPK pathway activation. An inhibitor of NLRP3 inflammasomes (MCC950) or an inhibitor of the p38 MAPK pathway (SB203580) further enhanced the ameliorative effects of PNS and BSP. PN-BS reduces esophageal barrier dysfunction by inhibiting the activation of NLRP3 inflammasomes and p38 MAPK pathway, thereby improving RE.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00858-9.

The imbalance of T helper 17 (Th17)/regulatory T (Treg) is pivotal in the development of chronic obstructive pulmonary disease (COPD). This study focused on evaluating the regulatory effects of short-chain fatty acids (SCFAs) on Th17/Treg balance in COPD. A COPD mouse model was induced by exposure to cigarette smoke (CS) and intranasal lipopolysaccharide (LPS) instillation. Mice were administered sodium acetate (NaA), sodium propionate (NaP), or sodium butyrate (NaB) via oral gavage. Lung function, histopathological changes, cytokine levels, and the distribution of Th17 and Treg cells were then evaluated. Epithelial cell injury induced by cigarette smoke extract (CSE) was assessed using the Cell Counting Kit-8 (CCK-8) assay, wound healing assay, and flow cytometry for apoptosis and T cell subset analysis. SCFAs treatment attenuated pathological damage in lung tissue, including reduced inflammatory cell infiltration and pulmonary fibrosis. SCFAs suppressed apoptosis and increased peak inspiratory flow (PIF) and peak expiratory flow (PEF). SCFAs effectively rebalanced the Th17/Treg axis by suppressing Th17 differentiation while promoting Treg cell expansion, which was accompanied by reduced levels of inflammatory signaling factors (IL-17, IL-6, IL-2, and TNF-α). Moreover, SCFAs enhanced CSE-induced MLE-12 cell viability and migration, and suppressed apoptosis. SCFAs blocked the differentiation of naïve CD4⁺ T cells into Th17 cells while facilitating their differentiation into Treg cells. SCFAs alleviate the pathological features of COPD by restoring the Th17/Treg balance and enhancing epithelial resilience, suggesting their promise as a therapeutic strategy for CS-related COPD.