MF VSI Chirurginės gastroenterologijos laboratorija (04.06.01)

Focal nodular hyperplasia (FNH) is a benign liver tumor commonly found in women, often linked to oral contraceptive use. Its occurrence in men is rare and frequently mimics malignant liver lesions, complicating diagnosis and management. This systematic review aims to evaluate the clinical presentation, diagnostic challenges, and treatment approaches of FNH in male patients.

Tikslas. Įvertinti imuninio atsako, susijusio su periferinio kraujo vienbranduolėmis ląstelėmis (PKVL), pokyčius po chirurginio kiaušidžių vėžio (KV) gydymo. Metodai. Atliktas perspektyvusis stebimasis tyrimas, į kurį įtrauktos 23 pacientės, sergančios III-IV stadijos КV ir kurioms atlikta citoredukcinė operacija, ir 23 kontrolinės grupės moterys. Tirta periferinio kraujo ląstelių sudėtis, PKVL subpopuliacijos, jų funkcinis aktyvumas, citokinų raiška PKVL ir jų kiekis serume. Rezultatai. KV sergančioms pacientėms prieš operaciją nustatytas sumažėjęs CD8+ T ląstelių kiekis, padidėjusi М2 monocitų dalis ir pakitusi citokinų raiška. Ankstyvuoju laikotarpiu po operacijos šie imuninės sistemos slopinimo požymiai išliko, be to, sumažėjo PKVL metabolinis aktyvumas ir fagocitozė bei padidėjo IL-6 kiekis serume. Išvados. KV sergančių pacienčių imuninės sistemos pokyčiai rodo galimą priešnavikinio imuniteto slopinimą, kuris išlieka ankstyvuoju laikotarpiu po operacijos.

Background/Objectives: Early and accurate detection of postoperative complications (POCs) remains a major challenge in colorectal cancer (CRC) surgery, underscoring the need for reliable molecular biomarkers. This study evaluated whether plasma miR-21 and miR-181a can predict POCs following left-sided CRC resection. Methods: This longitudinal observational sub-study was conducted within a randomized controlled trial. Adult patients undergoing elective left-sided CRC resection were included. Plasma miR-21 and miR-181a levels were measured preoperatively and on postoperative day (POD) 6 using RT-qPCR. POCs were assessed according to Clavien–Dindo classification. Of 40 enrolled patients, 38 were included in the final analysis (15 with and 23 without postoperative complications). Discriminative performance was assessed using receiver operating characteristic analysis and correlations with inflammatory markers were evaluated. Results: No significant differences in plasma miR-21 or miR-181a levels were observed between groups at baseline or POD6 (all p > 0.05). Both biomarkers showed limited discriminative ability (AUC = 0.61 and 0.54, respectively), while a combined model of miR-181a + TNF-α improved performance (AUC = 0.76, 95% CI: 0.57, 0.94, p = 0.01). At baseline, miR-21 correlated strongly with miR-181a (ρ = 0.81, p < 0.001) and moderately inversely with TNF-α (ρ = −0.35, p = 0.043). Conclusions: MiR-21 or miR-181a measured at baseline or POD6 show limited predictive value for POCs after CRC surgery. Further studies would benefit from larger sample sizes and optimized sampling strategies that reflect possible early dynamic changes in these biomarkers.

Background/Objectives.
Pancreatic ductal adenocarcinoma (PDAC) is one of the most fatal malignancies due to its aggressive nature and resistance to therapy. The epithelial-mesenchymal transition (EMT) drives cancer progression, regulated by transcription factors (TFs) such as SNAI1, SNAI2, ZEB1, ZEB2, and TWIST. This study evaluates EMT-TF expression in PDAC and its clinical relevance.
Methods.
PDAC tissues from 45 patients were analyzed using qRT-PCR and Western blot. Clinical features and survival outcomes were statistically examined for correlations with EMT-TF levels.
Results.
mRNA levels of SNAI1 (16.4-fold, p = 0.02), SNAI2 (21.8-fold, p = 0.028), ZEB1 (17.2-fold, p = 0.037) were significantly elevated in PDAC tissues compared to healthy controls. TWIST showed 3.75-fold increase in PDAC tissue; however, this elevation was not significant (p = 0.124). Corresponding protein-level increases were observed for Snail1/Slug and Zeb1. High SNAI1 expression correlated with peripancreatic invasion (p = 0.026) while ZEB1 overexpression was significantly associated with shorter survival (15.2 vs. 33.3 months, p = 0.037) and remained an independent prognostic factor in multivariate analysis. ZEB2 mRNA was reduced, however, protein levels were elevated. TWIST mRNA overexpression was not reflected at the protein level. Conclusions.
Overexpression of EMT transcription factors SNAI1 and ZEB1 reflect more aggressive histopathological patterns of PDAC. The strong correlation of SNAI1 expression with the expression of other EMT-TFs highlights its’ role in peripancreatic invasion, as well as impact on overall survival and may serve as an argument defining the leading role of SNAI1 in this context.

Leidinyje pateikiamos rekomendacijos, reglamentuojančios baigiamojo magistro darbo (BMD) rengimo procesą Medicinos vientisųjų studijų programoje. Dokumentas apibrėžia darbo planavimo, tyrimų etikos, metodologinių reikalavimų ir mokslinės komunikacijos principus, užtikrinančius akademinę kokybę ir tyrimų patikimumą.

Pancreatic ductal adenocarcinoma (PDAC) remains largely unresponsive to immunotherapy because of its highly immunosuppressive tumor microenvironment. Aryl hydrocarbon receptor (AHR), a ligand-dependent transcription factor, has emerged as a key regulator of immune homeostasis and inflammation. However, its systemic immunomodulatory role in PDAC, particularly outside the tumor microenvironment, remains poorly understood.

Aryl hydrocarbon receptor (AHR) is a transcription factor that’s commonly upregulated in pancreatic ductal adenocarcinoma (PDAC). There are evidence that upregulated AHR increases PDAC aggressiveness, invasiveness, migration and decreases survival rates. This can be linked to epithelial-mesenchymal transition, when cells acquire a migratory mesenchymal phenotype. AHR expression levels have been shown to influence epithelial or mesenchymal phenotype in some cancer types, however little is known how targeting AHR would affect cell migration and invasiveness in pancreatic cancer. The aim of this study was to investigate how targeting AHR would affect cell migration and invasiveness in pancreatic cancer cells. Two PDAC cell were used for the study (BxPC-3 and Su.86.86). AHR was silenced by lipofectamine mediated siRNA transfection, inhibited by AHR inhibitor (BAY compound) or knocked-out by using CRISPR-CAS system. After silencing, inhibiting or knocking-out AHR, scratch assay was performed to asses the effect on cell migration in 2D system. AHR protein inhibition influence on cell invasiveness and migration was also assessed in 3D system by growing them in low-adherence plates until they formed spheroids. The spheroids were then moved into adherence allowing plates to asses migration. Alternatively the spheroids were put into an extracellular matrix to asses invasion. The results showed that targeting AHR decreases PDAC cells migration and invasiveness. 2D Scratch assay showed that the migration was decreased by silencing AHR (BxPC-3 by 73 %; Su.86.86 by 54 %), inhibiting (BxPC-3 by 36 %; Su.86.86 – no change) and knocking-out (BxPC-3 by 51 %; Su.86.86 – KO not tested). 3D spheroid assays revealed similar trends of reducing cell migration (by 33-41 % depending on cell line) and invasiveness (by 20-43 % depending on cell line) after inhibiting AHR with BAY compound. Targeting AHR could prove to be a viable strategy in slowing PDAC progression by reduce cell invasiveness and migratory capabilities, however differences between cell lines suggest that such strategy should be pursued as a personalized treatment with other molecular mechanisms in mind.

Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer-related deaths, with a five-year survival rate below 10%. Its aggressiveness and chemotherapy resistance are linked to epithelial-mesenchymal transition (EMT), a process enabling epithelial cells to gain mesenchymal properties, enhancing migration and metastasis. EMT is driven by transcription factors such as SNAIL, SLUG, ZEB1/2, and TWIST. SNAI1 plays a key role in metastasis, while high ZEB1 expression correlates with poor survival. This process is tightly regulated by signalling networks, including ELAVL1, which encodes HuR, a protein that stabilizes mRNAs by binding to AU-rich elements (AREs) in their 3′ untranslated regions (3′UTRs), enhancing translation. Tumor tissues from 65 PDAC patients undergoing surgical resection were analyzed. Total RNA was extracted and converted into cDNA for qRT-PCR. BxPC-3, MiaPaCa-2, and Su.86.86 cell lines were cultured in RPMI medium with 10% fetal bovine serum and 1% antibiotics at 37°C in a 5% CO2 humidified environment. For immunoprecipitation, 1–2 × 107 cells were lysed using a manufacturer-provided protocol. Mouse monoclonal anti-HuR (ELAVL1) antibody was used for protein binding, with normal mouse IgG as a control. Samples were analyzed via qRT-PCR. Binding sites were annotated using the CISBP-RNA database, considering only RNAdirect-confirmed 3′UTR motifs. Target gene 3′UTR sequences were retrieved from GENCODE V47 (GRCh38). Statistical analysis was performed using GraphPad Prism, applying the Kruskal-Wallis test with Dunn’s multiple comparisons and Spearman’s correlation. Data were presented as median with ± interquartile range, with significance at p<0.05. A strong positive correlation was identified between ELAVL1 and the transcription factors ZEB1, SNAI1, and SNAI2 (r = 0.74, 0.76, and 0.76, respectively). Expression levels of EMT-TFs and ELAVL1 were classified as high and low. Elevated ELAVL1 expression was associated with a 71.65-fold increase in ZEB1, whereas high ZEB1 levels corresponded to a 96.1% reduction in ELAVL1 expression. Similarly, high ELAVL1 levels resulted in a substantial upregulation of SNAI1 (312.35-fold), while even low ELAVL1 expression maintained elevated SNAI1 levels (8.45-fold). However, when SNAI1 expression was high, ELAVL1 levels declined by 90%, whereas simultaneous high expression of both factors led to a moderate 3.71-fold increase in ELAVL1. High SNAI2 expression was associated with a 93% reduction in ELAVL1, whereas low SNAI2 levels corresponded to a striking 198.11-fold increase in ELAVL1. Structural analysis identified multiple ELAVL1 binding sites: 34 in ZEB1, 1 in SNAI1, and 9 in SNAI2. Immunoprecipitation analysis confirmed that ELAVL1 can potentially bind to ZEB1, SNAI1, and SNAI2 in MiaPaCa-2 and Su.86.86 cell lines. In conclusion, our findings highlight a significant regulatory interplay between ELAVL1 and key EMT transcription factors in PDAC. The strong positive correlation between ELAVL1 and these genes, along with their observed expression patterns, suggests that ELAVL1 plays a crucial role in EMT regulation. Furthermore, structural mapping identified multiple ELAVL1 binding sites within these genes, reinforcing its post-transcriptional regulatory function. These insights lay the groundwork for further investigations into ELAVL1 as a potential therapeutic target in EMT-driven pancreatic cancer progression.

Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest cancers, partly because the cancer cells can evade the immune system, also the pancreatic cancer cells are highly heterogeneous, and this tumor has a late onset of symptoms, which become visible only in advanced stages. For this study, we wanted to investigate the role of the aryl hydrocarbon receptor (AhR), a potential regulator of immune response and inflammatory pathways in PDAC. Using patient-derived peripheral blood mononuclear cells (PBMCs), we sought to determine how modulation of AhR activity affects the expression and production of immune checkpoint molecules and inflammatory mediators. To reflect the subtle effects of AhR activity on immunity, we divided PBMCs from 22 PDAC patients into four groups: unmodulated control, AhR stimulated by carbidopa, AhR stimulated by tapinarof, and AhR suppressed by bay. After 24 hours of modulation, the PBMCs were frozen for gene expression analysis using RT-PCR. The culture supernatants were also frozen for target protein analysis using ELISA. After experimental analysis all patient results were separated into two parts based on the AhR expression level of the unmodulated group: low AhR and high AhR. The Mann-Whitney test was used to determine the statistical significance in GraphPad software. Our study demonstrated different patterns of gene regulation in response to AhR modulation. In the high AhR expression group, carbidopa significantly downregulated AhR expression (p < 0.05), while in the low AhR expression group, tapinarof induced a significant upregulation of AhR (p < 0.05). Regardless of baseline AhR levels, tapinarof stimulation consistently increased CYP1A1 transcription (p < 0.05) across both groups. Conversely, AhR inhibition with bay resulted in a decreasing trend of CYP1A1 expression in both AhR groups. Interestingly, PTGS2 (encoding COX-2) remained downregulated in AhR-stimulated groups, but no reliable changes were observed. AhR modulation also affected immune checkpoint gene expression. Tapinarof treatment showed an upregulating trend in CD274 (PD-L1) transcription, independent of the initial AhR levels, while CD279 (PD-1) expression remained relatively stable across all conditions. At the protein level, carbidopa treatment was associated with a decreasing trend in soluble PD-L1 concentrations across both AhR expression groups. In contrast, soluble PD-1 levels remained elevated following AhR stimulation with carbidopa and tapinarof. Notably, soluble PGE2 levels showed a decreasing trend in the low AhR group after suppression with bay. These findings suggest the unique immunological role of AhR primary expression, functional activity, and different modulators in PDAC patients' blood. AhR activation enhances CYP1A1 expression, slightly reduces PD-L1 protein production with carbidopa, and affects inflammation-related genes in a context-dependent manner. AhR inhibition attenuates these responses, particularly in patients with low baseline AhR expression. However, these results suggest that AhR may be one of the main targets as a key regulator of immune responses in PDAC, and present new strategies for improving the efficacy of immunotherapeutic interventions.

Background and Objectives: Fireweed (Chamerion angustifolium L.) possesses antibacterial, antioxidant, anti-inflammatory, and anti-cancer properties. This study evaluated the effects of aqueous fireweed leaf extracts and their major compound, oenothein B, on the viability and mitochondrial function of Caco-2 colon cancer cells, emphasizing the impact of leaf fermentation. Materials and Methods: Cells were treated for 48 h with oenothein B and aqueous extracts from unfermented (NF) and fermented leaves (F 24 h, F 48 h). Cell viability and mitochondrial function were assessed by MTT assay and high–resolution respirometry. Results: IC50 values were 0.843 mg/mL (NF), 1.548 mg/mL (F 24 h), 1.931 mg/mL (F 48 h), and 0.09 mg/mL (57 µM) (oenothein B). Mitochondrial respiration decreased in up to 67% (glutamate/malate) and 61% (succinate) in both fermented and unfermented groups, while oenothein B increased in leak respiration by 34–73% but reduced oxidative respiration by 24%. Conclusions: Aqueous extracts of fireweed from both unfermented and fermented leaves significantly reduced Caco-2 cell viability and mitochondrial function. Oenothein B on its own had a stronger effect on cell viability, but a weaker effect on mitochondrial function, compared to fireweed extracts.