Search Results (833)

Given its highly aggressive nature and poor clinical outcome, pancreatic ductal adenocarcinoma (PDAC) requires physiologically relevant in vitro models that more accurately reflect tumor biology and drug response. In this study, adhesive and non-adhesive hydrogel microenvironments were comparatively evaluated for pancreatic cancer spheroid modeling using PANC-1 and MIA PaCa-2 cells. Gelatin methacryloyl (GelMA) hydrogels were synthesized, photocrosslinked, and optimized in terms of stability, swelling, degradation, and cytocompatibility, while 3% agarose was used as a non-adhesive counterpart. Although the optimized GelMA formulation showed adequate structural stability and no cytotoxicity, it did not support spheroid formation. In contrast, agarose enabled the formation of compact, viable, and proliferative spheroids in both cell lines. Agarose-derived spheroids exhibited time-dependent growth, positive Ki-67 staining, and increased HIF-1α expression under 3D conditions, indicating the establishment of hypoxia-associated tumor-like microenvironments. Gemcitabine treatment induced a time-dependent reduction in spheroid viability, while viable cell populations persisted throughout exposure, reflecting the heterogeneous therapeutic response typical of 3D tumor models. Overall, these findings provide a comparative, microenvironment-based assessment of pancreatic cancer spheroid modeling, indicating that hydrogel-dependent differences in adhesivity and structural dynamics are important determinants of spheroid assembly, hypoxia-associated molecular adaptation, and chemotherapeutic response. Overall, these findings provide a comparative, microenvironment-based assessment of pancreatic cancer spheroid modeling, indicating that hydrogel-dependent differences in adhesivity and structural dynamics are important determinants of spheroid assembly, hypoxia-associated molecular adaptation, and chemotherapeutic response. Overall, these findings provide a comparative, microenvironment-based assessment of pancreatic cancer spheroid modeling, indicating that hydrogel-dependent differences in adhesivity and structural dynamics are important determinants of spheroid assembly, hypoxia-associated molecular adaptation, and chemotherapeutic response. Full article

Enteroviruses are positive-sense single-stranded RNA viruses and common pathogens that are responsible for diverse public health diseases. To facilitate the study of the virus biology and pathogenesis of enterovirus, we developed a rapid method for construction of the enteroviral cDNA clones including enterovirus A71 (EV-A71) and coxsackievirus B5 (CVB5). As described for EV-A71, the full-length cDNA of CVB5 was amplified by long-distance PCR and cloned into a T7 promoter-containing plasmid using directional seamless cloning technology. The virus was successfully rescued by single transfection into cells stably expressing T7 polymerase and exhibited characteristics similar to the parental virus. Next, through systematic construction and the optimization of the EV-A71 and CVB5 reporter viruses, we successfully generated two novel reporter virus panels with high virus titers, rapid replication, and relatively stable genetic inheritance across passages using the new fluorescence proteins mScarlet3-H and the smallest miRFP670nano3. Analysis of critical determinants for the reporter virus construction revealed that reporter gene sizes, genomic insertion sites, and the usage of protease recognition sites are crucial parameters. The EV-A71 and CVB5 reporter viruses enable antiviral drug evaluation, as demonstrated by our identification of gemcitabine as a broad-spectrum inhibitor of both viruses. These systems also facilitate the functional interrogation of host factors, exemplified by our discovery that METTL3 promotes EV-A71 and CVB5 replication. These reverse genetic tools, including infectious cDNA clones and reporter viruses, will advance basic enterovirus biology and accelerate antiviral drug discovery. Full article

Upper tract urothelial carcinoma (UTUC) is a rare and aggressive malignancy, accounting for only 5–10% of all urothelial carcinomas (UCs). Lung, bone, liver, and distant lymph nodes are common sites of metastasis, while gastrointestinal metastasis is extremely rare. We present a case of a 63-year-old female who developed a descending colon lesion 19 months after left radical nephroureterectomy for high-grade ureteral UC. The diagnosis was established by computed tomography (CT), magnetic resonance imaging (MRI), colonoscopy, and biopsy, which excluded primary colorectal malignancy. First-line therapy consisted of six 21-day cycles of gemcitabine plus cisplatin, followed by two cycles of tislelizumab maintenance immunotherapy. Restaging with contrast-enhanced CT and positron emission tomography/computed tomography (PET/CT) demonstrated disease progression. Despite switching to second-line nab-paclitaxel, the patient rapidly deteriorated from tumor cachexia and ultimately succumbed to septic shock secondary to severe pulmonary infection. This represents the first reported case of descending colon metastasis from primary ureteral UC. It highlights the colon as a potential metastatic site where biopsy is essential for definitive diagnosis. Notably, although the patient initially responded to platinum-based therapy, the subsequent rapid progression underscores the need for vigilant monitoring and timely adjustment of therapeutic strategies in managing such high-risk presentations. Full article

Pharmacological inhibition of the nucleoside transporter hENT1 is a promising therapeutic target across a range of diseases, including cardiovascular disorders, neurodegenerative conditions, and cancer. However, current inhibitors lack drug-like properties, necessitating the development of new inhibitors with improved pharmacological profiles. We employed a dual-pharmacophore virtual screening protocol to identify putative hENT1 inhibitors from a library of over 2 million compounds, followed by structure-based molecular docking. To validate the inhibition effect of the lead compounds, we established a functional assay using gemcitabine (GEM)-induced cytotoxicity as a readout of hENT transport activity using eight cancer cell lines. H292 was the optimal cancer cell line for the validation assay based on its high GEM sensitivity (IC50 = 28 nM) and the concentration-dependent cytotoxicity inhibition of the reference inhibitor NBTI, a hENT1 inhibitor. Of the 19 candidate compounds, two leads (compounds 2 and 3) demonstrated potency comparable to NBTI, increasing GEM IC50 values by 2.2- and 2.9-fold at 5 µM, respectively. Both compounds were non-cytotoxic to normal fibroblasts, exhibited favorable ADME properties, displayed superior docking scores of −12.63 and −12.49 kcal/mol compared to NBTI (−9.06 kcal/mol), and displayed a novel vertical binding orientation within the hENT1 binding pocket distinct from NBTI’s horizontal mode. This study established a validated non-radioactive, gemcitabine-based functional assay for hENT inhibitor discovery and identified two putative inhibitors with therapeutic potential for cancer chemosensitization, pain management, and cardio- and neuroprotection. The non-radioactive functional assay overcomes the limitations of traditional radiolabeled methods, enabling scalable, broader screening applications. Full article

While artemisinin and its derivatives demonstrate broad antitumor potential, the stereochemical influence on the bioactivity of higher-order artemisinin assemblies remains inadequately characterized. Herein, we report the synthesis, chromatographic separation, and structural elucidation of four stereoisomeric artemisinin trimers, followed by systematic evaluation of their antitumor efficacy against MCF-7 and MDA-MB-231 breast cancer cell lines. All trimers exhibited potent cytotoxicity against MCF-7 cells (IC₅₀ < 0.09 μM), with trimer 6a (β, β, β) demonstrating robust antitumor activity in both in vitro and in vivo xenograft models. Remarkably, pronounced stereochemistry-dependent activity emerged against MDA-MB-231 cells: 6a displayed approximately 100-fold greater potency than 6b (β, β, α) and 6.6-fold superiority over gemcitabine. Mechanistic investigations revealed that 6a downregulates Cyclin D1, CDK4, and CDK6 expression, thereby inducing G0/G1 phase cell cycle arrest. These findings underscore the pivotal role of stereochemical configuration in modulating artemisinin trimer bioactivity and provide rational guidance for structure-based design of artemisinin-derived anticancer therapeutics. Full article

Introduction: Lung cancer remains the leading cause of cancer-related mortality among men in Poland. Prognosis is generally poor, largely due to late diagnosis at advanced stages and the aggressive biological nature of the disease. Aim: This study aimed to evaluate the effectiveness of various treatment modalities and determine their impact on overall survival in male patients diagnosed with small-cell (SCLC) and non-small-cell lung cancer (NSCLC). Methods: This retrospective cohort study analyzed 1431 men (mean age: 61.5 years) treated at the Katowice Oncology Center in Poland between 2002 and 2012. Overall survival was assessed using the Kaplan–Meier method and multivariable Cox proportional hazards regression. Evaluated prognostic factors included clinical stage, surgical intervention (partial or total lung resection), first-line treatment regimen, and the number of treatment cycles. Results: Survival probabilities declined progressively with advancing clinical stage for both SCLC and NSCLC. Patients who underwent surgical resection demonstrated significantly longer survival compared to non-surgically treated patients (p < 0.001). Furthermore, combined radiochemotherapy yielded superior therapeutic outcomes compared to chemotherapy alone. In the non-surgical NSCLC cohort, first-line treatment with platinum derivatives combined with gemcitabine resulted in the highest 1-year survival rate compared to other pharmacological schemes. Discussion: The high mortality observed within the first 12 months post diagnosis reflects the late-stage presentation common during the study period. The findings align with established oncological principles, confirming that surgical resection and multimodal therapies offer the greatest survival advantages for eligible patients. Conclusions: Survival rates for both SCLC and NSCLC are overwhelmingly dictated by early diagnosis and the feasibility of surgical resection. Improving long-term outcomes depends heavily on implementing effective lung cancer screening programs to detect the disease at operable stages and utilizing optimized combined treatment protocols. Full article

Cholangiocarcinoma (CCA) is a highly lethal, heterogeneous malignancy arising from the biliary tract. Although the prevalence of CCA is relatively low, its incidence has increased in the last few decades, and the overall prognosis is poor. Surgical resection remains the most efficacious treatment modality for CCA. However, due to its aggressive nature and often asymptomatic presentation, most patients are first diagnosed with advanced disease, precluding them from curative intervention. Moreover, due to its heterogeneity at the molecular, genomic, and epigenetic levels, drug treatment of CCA remains challenging. In this review, we discuss the current standard drug treatment approaches, recent breakthroughs, and promising new therapeutics for CCA. We summarize key clinical data for the standard first-line chemotherapy regimen and its efficacy and resistance mechanisms, along with more recent studies supporting or proposing second-line treatments. We highlight landmark clinical trials, including ABC-02, which established gemcitabine-cisplatin (GC) as the first-line regimen against biliary cancers. Additionally, we discuss recent findings on the susceptibility of CCA against targeted therapies and other immunologic molecules, including results from the KEYNOTE-966 and TOPAZ-1 clinical trials. Finally, we critically analyze new therapeutics in the preclinical and clinical space, such as CAR-T cells and oncolytic viruses that may be effective against CCA. Full article

Atractylodes lancea (AL) has been shown to be a promising candidate for the treatment of cholangiocarcinoma (CCA). The study explored the potential of atractylodin (AT) and β-eudesmol (BE) to chemosensitize the effects of standard chemotherapeutics in CCA. The cytotoxicities of AT and BE on CL6, HuCCT1, and HuH28 when used in combination with 5-fluorouracil (5FU), gemcitabine (GEM), and cisplatin (Cis) were assessed by MTT assay. The modulatory effects of both compounds on mRNA expression of the reuptake and efflux transporters were determined by real-time PCR. The FIC (Fractional Inhibitory Concentration) indices indicated synergistic interactions (AT-5FU in all cell lines and BE-5FU in HuH28) and antagonistic interactions (BE-Cis in all cell lines and AT-Cis or AT-GEM in HuCCT1). The synergistic interactions observed with the AT-5FU and BE-5FU combinations were well correlated with the significant upregulation of the mRNA expression of the reuptake transporter genes hENT1 (2.64-fold) and hOCT3 (5.02-fold) and the significant downregulation of the mRNA expression of the efflux transporter gene ABCC2 (0.33-fold). AT and BE, when purified or present as significant components in AL, may benefit CCA treatment when used as adjunct therapy to standard chemotherapeutic drugs, particularly 5FU. The mechanism of synergistic activity may, at least in part, involve modulation of transporter gene expression and activity. Full article

Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal malignancy marked by substantial molecular heterogeneity and variable response to gemcitabine-based therapy. While KRAS mutations are nearly universal, the broader RTK-RAS and MAPK signaling architecture and its relationship to treatment response remain incompletely defined. We conducted an integrative clinical-genomic analysis of 184 PDAC tumors stratified by age at diagnosis and gemcitabine exposure, interrogating somatic alterations across curated RTK-RAS/MAPK gene sets. Conversational artificial intelligence agents (AI-HOPE-RTK-RAS and AI-HOPE-MAPK) enabled dynamic cohort construction and pathway-level analyses, with findings validated using standard statistical methods. In late-onset PDAC, ERBB2 and RET mutations were significantly enriched in gemcitabine-treated tumors. Early-onset cases demonstrated differential enrichment of CACNA2D family alterations in non-treated tumors and higher frequencies of FLNB and TP53 mutations in treated disease. Importantly, late-onset patients not treated with gemcitabine who lacked RTK-RAS or MAPK alterations exhibited significantly improved overall survival. These findings reveal age- and treatment-dependent pathway dependencies beyond canonical KRAS status and support a precision oncology framework in PDAC. Conversational AI facilitated rapid, multidimensional clinical–genomic integration to uncover clinically relevant signaling substructures. Full article

Bladder cancer (BCa) is the second most common cancer of the genitourinary tract globally. It has limited treatment options, high recurrence rate, and acquires resistance to platinum-based therapy. Therefore, identifying novel therapeutic targets is urgently needed. Analysis of the TCGA data revealed that the enzyme galactokinase-1 (GALK1) is overexpressed (p < 0.0001) in bladder tumors compared to normal tissue. Our data also confirmed GALK1 protein upregulation in multiple human BCa cell lines and rodent bladder tumors. However, the precise role of GALK1 in BCa progression and effects of its specific inhibitor remain unexamined. In this study, we demonstrate that GALK1 gene silencing using shRNA resulted in a significant reduction in BCa cell proliferation, migration, and invasion. Pharmacological inhibition of GALK1 using small molecule Cpd36 resulted in anticancer efficacy against BCa. Cpd36 inhibited proliferation, migration, and invasion of BCa cells. Further, Cpd36 induced G1 phase cell cycle arrest, apoptosis, mitochondrial membrane depolarization, and ROS production in the BCa cells. Mechanistically, Cpd36-induced reduction in cell proliferation was associated with a decrease in expression of GALK1, PCNA proteins. Inhibition of metastatic potential was accompanied by decreased migration, invasion, and MMP-9 expression. Cell cycle arrest was associated with decrease in Cyclin D1 and increased expression of p21 and p27. Induction of apoptosis was linked with increased expression of cleaved caspase-3 and cleaved PARP, while downregulating p-AKT. Additionally, Cpd36 in combination with cisplatin or gemcitabine showed a strong synergistic effect on BCa cells. Taken together, our findings suggest that GALK1 plays a significant role in BCa cell survival and validates its inhibitors as promising therapeutic options for managing this disease. Full article

Background: The primary aim of this Phase 1 clinical trial was to study the safety and dose of a cholecystokinin receptor antagonist, proglumide, in combination with gemcitabine/nab-paclitaxel (GEM-NAB-P) in patients with metastatic pancreatic cancer. The secondary aim was to study the effects of proglumide with GEM-NAB-P on the tumor microenvironment (TME) with tumor biopsies and a blood biomarker assay. An exploratory aim studied the effects of proglumide treatment on cancer-related pain. Methods: Gemcitabine-naïve patients were treated with GEM-NAB-P plus proglumide 1200 mg/day. Tumor biopsies and a liquid biopsy serum sample for analysis of a microRNA biomarker panel were collected pre- and on-treatment to study the TME. McGill pain surveys were done at baseline, week 8 and at the end of treatment. The study was approved and registered (NCT05827055). Results: The mean age of the patients was 68.2 years (range 54–74 years). The starting dose was well-tolerated with no unexpected treatment-related adverse events observed. Multiplex immunohistochemical analysis of tumor biopsies at baseline and week 8 revealed a significant reduction in Ki67+ cells, collagen1α1, and M2-polarized tumor-associated macrophages (TAMs). Week 8 tumor biopsies demonstrated a significant increase in CD8+ T-cells and natural killer cells compared to baseline. The blood biomarker panel showed a significant inverse change in microRNAs associated with decreasing fibrosis and metastasis. The McGill pain scores showed less pain at week 24 or end-of-treatment compared to baseline. Conclusions: Proglumide demonstrates a favorable safety profile when combined with standard chemotherapy for metastatic pancreatic cancer. Its unique ability to remodel TME and alleviate cancer-related pain highlights its potential, warranting further research. Full article

Background/Objectives: The present study aimed to compare the efficacy and safety of intravesical gemcitabine versus Bacillus Calmette–Guérin administered after transurethral resection of bladder tumor in patients with non-muscle-invasive bladder cancer. Methods: A comprehensive literature search was conducted using PubMed, Embase, and the Cochrane Library databases up to September 2025. Studies were eligible for inclusion if they were comparative studies evaluating intravesical gemcitabine versus Bacillus Calmette–Guérin following transurethral resection of bladder tumor. The primary outcomes were recurrence-free survival, progression-free survival, and incidence of adverse events. Pooled hazard ratios were calculated using a random-effects model based on log hazard ratios and standard error values. Results: Seven studies comprising approximately 774 patients were included in this analysis. The pooled hazard ratio was 0.80 (95% confidence interval, 0.42–1.53) for recurrence-free survival and 0.76 (95% confidence interval, 0.46–1.26) for progression-free survival, indicating no significant difference between gemcitabine and Bacillus Calmette–Guérin. However, gemcitabine was associated with a significantly lower incidence of adverse events, with a pooled odds ratio of approximately 0.48 (95% confidence interval, 0.27–0.86). Risk-of-bias assessment revealed that most randomized trials had “some concerns” based on the Risk of Bias 2 tool, whereas non-randomized studies were rated as having “moderate to serious” risk of bias according to the Risk of Bias in Non-randomized Studies of Interventions tool. Conclusions: Gemcitabine demonstrated an oncologic efficacy comparable to that of Bacillus Calmette–Guérin in terms of recurrence and progression outcomes while showing a substantially lower incidence of treatment-related toxicities. Full article

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy with a 5-year survival rate of 13.3%. First-line treatment relies on two chemotherapy regimens, FOLFIRINOX (FOLFNX) or gemcitabine plus nab-paclitaxel (GEMPAC). However, direct clinical comparisons between these regimens have yielded inconsistent results across survival and toxicity endpoints, and the molecular basis of heterogeneous treatment responses remains poorly defined. To investigate regimen-specific tumor-cell-intrinsic mechanisms, we performed quantitative proteomic profiling of a primary PDAC-derived MIA PaCa-2 cell line following treatment with FOLFNX or GEMPAC. Differentially expressed proteins were analyzed using Gene Ontology, KEGG, and Ingenuity Pathway Analysis to define pathway-level alterations, and findings were contextualized using TCGA transcriptomic data. Proteomic analyses revealed that FOLFNX and GEMPAC engage in distinct cytotoxic programs. FOLFNX predominantly suppressed ribosome biogenesis and mitochondrial translation, consistent with sustained metabolic and biosynthetic stress, whereas GEMPAC preferentially disrupted mitotic cytokinesis and phosphatidylinositol phosphate biosynthesis, consistent with mitotic failure. Integration with TCGA data showed that FOLFNX-altered proteins aligned with favorable prognostic expression signatures, whereas GEMPAC-associated proteins were enriched among adverse profiles, reflecting engagement of distinct tumor-intrinsic programs. Together, these findings provide mechanistic insight into differential chemotherapy responses and establish a foundation for proteomics-based biomarkers to guide personalized chemotherapy selection in PDAC. Full article

Background: Treatment decisions for older adults with locally advanced or metastatic pancreatic ductal adenocarcinoma (PDAC) often rely on heterogeneous observational evidence and clinical judgment regarding survival benefits, regimen intensity, and tolerability. Methods: We systematically searched Embase, PubMed, and Scopus from inception to 30 March 2025, for studies reporting overall survival (OS) and/or progression-free survival (PFS) in older adults with advanced PDAC receiving systemic chemotherapy, as well as age-stratified outcomes among chemotherapy-treated patients. Hazard ratios (HRs) with 95% confidence intervals (CIs) were primarily extracted from multivariable-adjusted analyses. In cases without reported HRs, estimates were derived from summary statistics or Kaplan–Meier curves. The review protocol was registered in PROSPERO (CRD420261292913). Results: A total of 40 predominantly retrospective studies were included. Chemotherapy was associated with improved OS compared to best supportive care in older adults (9 studies; HR 0.46, 95% CI 0.39–0.54; I² = 18%). Among chemotherapy-treated patients, OS (34 studies; HR 1.00, 95% CI 0.99–1.02; I² = 23%) and PFS (11 studies; HR 0.96, 95% CI 0.86–1.07; I² = 10%) did not differ by age. Combination chemotherapy demonstrated superior OS (13 studies; HR 0.66, 95% CI 0.54–0.80; I² = 86%) with substantial heterogeneity and PFS (7 studies; HR 0.63, 95% CI 0.53–0.74; I² = 30%) compared to monotherapy. FOLFIRINOX and gemcitabine plus nab-paclitaxel demonstrated comparable OS (8 studies; HR 0.98, 95% CI 0.90–1.05; I² = 60%) and PFS (2 studies; HR 0.97, 95% CI 0.92–1.02; I² = 0%). Conclusions: Among carefully selected older adults with advanced PDAC, chemotherapy was associated with improved survival compared to supportive care. Chronological age did not predict outcomes, highlighting the need for geriatric-informed prospective trials. Full article

Leucine-rich repeat-containing 8A (LRRC8A; also known as SWELL1), the essential subunit of volume-regulated anion channels (VRACs), is amplified in multiple malignancies and has been implicated in tumor progression and therapeutic resistance. Three-dimensional (3D) cancer spheroids have been well-established as in vitro models that recapitulate characteristics of tumor stemness and intrinsic drug resistance. In the present study, spheroid formation in human breast cancer cell lines, YMB-1 and MDA-MB-468, conferred resistance to multiple anticancer drugs, including doxorubicin (DOX), gemcitabine (GEM), and 5-fluorouracil (5-FU), thereby mimicking the characteristic properties of breast cancer stem-like cells. LRRC8A expression was upregulated in 3D spheroids compared with adherent 2D monolayers, and its pharmacological inhibition induced membrane hyperpolarization accompanied by intracellular Cl⁻ accumulation. Inhibition of LRRC8A significantly sensitized spheroids to DOX, GEM, and 5-FU. Spheroid formation increased the expression of multidrug resistance-related protein 3 (MRP3) and the drug-metabolizing enzyme cytochrome P450 3A4 (CYP3A4), whereas LRRC8A inhibition suppressed their expression. The transcriptional upregulation of MRP3 and CYP3A4 was mediated through the NRF2–CEBPB/D transcriptional axis. Collectively, these findings suggest that LRRC8A inhibition may represent a therapeutic strategy to overcome chemoresistance by repressing MRP3 and/or CYP3A4 expression in breast cancer stem cells. Full article