Exosomes are essential for several tumor progression-related processes, including the epithelial–mesenchymal transition (EMT). Long non-coding RNAs (lncRNAs) comprise a major group of exosomal components and regulate the neoplastic development of several cancer types; however, the progressive role of exosomal lncRNAs in bladder cancer have rarely been addressed. In this study, we identified two potential aggressiveness-promoting exosomal lncRNAs, LINC00960 and LINC02470. Exosomes derived from high-grade bladder cancer cells enhanced the viability, migration, invasion and clonogenicity of recipient low-grade bladder cancer cells and activated major EMT-upstream signaling pathways, including β-catenin signaling, Notch signaling, and Smad2/3 signaling pathways. Nevertheless, LINC00960 and LINC02470 were expressed at significantly higher levels in T24 and J82 cells and their secreted exosomes than in TSGH-8301 cells. Moreover, exosomes derived from LINC00960 knockdown or LINC02470 knockdown T24 cells significantly attenuated the ability of exosomes to promote cell aggressiveness and activate EMT-related signaling pathways in recipient TSGH-8301 cells. Our findings indicate that exosome-derived LINC00960 and LINC02470 from high-grade bladder cancer cells promote the malignant behaviors of recipient low-grade bladder cancer cells and induce EMT by upregulating β-catenin signaling, Notch signaling, and Smad2/3 signaling. Both lncRNAs may serve as potential liquid biomarkers for the prognostic surveillance of bladder cancer progression.

Bladder cancer progression and metastasis have become major threats in clinical practice, increasing mortality and therapeutic refractoriness; recently, epigenetic dysregulation of epithelial-to-mesenchymal transition (EMT)-related signaling pathways has been explored. However, research in the fields of long noncoding RNA (lncRNA) and competing endogenous RNA (ceRNA) regulation in bladder cancer progression is just beginning. This study was designed to determine potential EMT-related ceRNA regulation in bladder cancer progression and elucidate the underlying mechanisms that provoke aggressiveness. After screening the intersection of bioinformatic pipelines, LINC02470 was identified as the most upregulated lncRNA during bladder cancer initiation and progression. Both in vitro and in vivo biological effects indicated that LINC02470 promotes bladder cancer cell viability, migration, invasion, and tumorigenicity. On a molecular level, miR-143-3p directly targets and reduces both LINC02470 and SMAD3 RNA expression. Therefore, the LINC02470–miR-143-3p–SMAD3 ceRNA axis rescues SMAD3 translation upon LINC02470 sponging miR-143-3p, and SMAD3 consequently activates the TGF-β-induced EMT process. In conclusion, this is the first study to demonstrate that LINC02470 plays a pivotally regulatory role in the promotion of TGF-β-induced EMT through the miR-143-3p/SMAD3 axis, thereby aggravating bladder cancer progression. Our study warrants further investigation of LINC02470 as an indicatively prognostic marker of bladder cancer.

Gemcitabine (GCB) resistance is a major issue in bladder cancer chemoresistance, but its underlying mechanism has not been determined. Epithelial-mesenchymal transition (EMT) has been shown to be comprehensively involved in GCB resistance in several other cancer types, but the direct connection between EMT and GCB remains unclear. This study was designed to elucidate the mechanism of EMT-related GCB resistance in bladder cancer and identify a potential phytochemical to modulate drug sensitivity. The biological effects of ellagic acid (EA) or its combined effects with GCB were compared in GCB-resistant cells and the GCB-sensitive line in terms of cell viability, apoptosis, motility, and in vivo tumorigenicity. The molecular regulation of EMT-related GCB resistance was evaluated at both the mRNA and protein expression levels. Our results indicated that TGF-β/Smad induced the overactivation of EMT in GCB-resistant cells and reduced the expression of GCB influx transporters (hCNT1 and hENT1). Moreover, ellagic acid (EA) inhibited the TGF-β signaling pathway both in vitro and in vivo by reducing Smad2, Smad3, and Smad4 expression and thereby resensitized GCB sensitivity. In conclusion, our results demonstrate that TGF-β/Smad-induced EMT contributes to GCB resistance in bladder cancer by reducing GCB influx and also elucidate the novel mechanisms of EA-mediated inhibition of TGF-β/Smad-induced EMT to overcome GCB resistance. Our study warrants further investigation of EA as an effective therapeutic adjuvant agent for overcoming GCB resistance in bladder cancer.

Extracellular vesicles (EVs) are critical mediators of intercellular communication within the tumour microenvironment and play a significant role in drug resistance. We aimed to investigate the mechanisms underlying gemcitabine (GCB) resistance in bladder cancer. GCB‐resistant bladder cancer cells exhibited dysregulation of nucleoside‐metabolizing enzymes and transporters. Characterization of EV subpopulations derived from GCB‐resistant cells revealed their ability to transfer drug‐resistant phenotypes to naïve cancer cells by modulating intracellular levels of nucleoside metabolic proteins and transporters. Proteomic and transcriptomic analyses identified the histone protein H3.2 and its corresponding transcript, H3C14, as key regulators in the transmission of GCB resistance. Notably, H3C14 overexpression in resistant cells restored GCB sensitivity, whereas its knockdown induced GCB resistance. Rab27A‐mediated biogenesis and secretion emerged as a crucial mechanism regulating EV release and H3C14 excretion in GCB‐resistant cells. A specific EV subpopulation enriched in CD147 and LAMB1—referred to as Excretion EVs—carried H3.2 (H3C14) but did not induce GCB resistance in recipient cells, suggesting their primary role in eliminating proteins associated with tumour progression and drug resistance. These findings highlight the role of EV‐mediated H3C14 excretion in regulating GCB resistance and suggest potential therapeutic strategies targeting EV pathways to overcome drug resistance in bladder cancer.

Objective: Long segment ureteral lesion with obstruction is a clinically difficult issue for recovering and maintaining organ or tissue function. Regeneration medicine using various biomaterials as a scaffold in supporting tissue regrowth is emerging. We developed this customized scaffold using electrospinning and 3-dimensional assistance and expected that it may provide an alternative biomaterial for ureter defect repair. Material and Methods: Our study synthesized polycaprolactone and silk fibroin combination as biomaterial scaffolds. The differences in physicochemical properties and biocompatibility of polycaprolactone–silk fibroin bio-scaffolds prepared by electrospinning alone and 3-dimensional printing combined with electrospinning in proper ratios were compared and characterized. SV-HUC-1 uroepithelial cells cultured in polycaprolactone–silk fibroin (4 : 6) scaffolds were observed under a scanning electron microscope and using calcein-acetomethoxy and propidium iodide stain. The ex vivo resected healthy human ureteral segment tissue was anastomosed with the polycaprolactone–silk fibroin scaffolds and cultured in an ex vivo bath for 2 weeks. The cellular growth on the polycaprolactone–silk fibroin scaffold was observed microscopically. In the New Zealand white rabbit model, we performed a 1/5 ratio (2 cm out of 10 cm) defect replacement of the unilateral ureter. After 7 weeks, the rabbits were sacrificed and the implanted ureter scaffolds were resected for tissue sectioning and the cellular growth was observed by hematoxylin and eosin and Masson staining. Results: When the proportion of silk fibroin was increased and the 3-dimensional electrospinning method was used, both the size and diameter of nanofiber holes were increased in the polycaprolactone–silk fibroin scaffold. Scanning electron microscope and fluorescent stain revealed that cultured 3T3 and SV-HUC-1 uroepithelial cells could electively penetrate inside the polycaprolactone–silk fibroin (4 : 6) nanofibrous scaffolds in 3 days. The polycaprolactone–silk fibroin scaffold anastomosis in an ex vivo bath showed cellular growth stably along the scaffold for 2 weeks, and most of the cells grow along with the outboard of the scaffold in layers. In an animal model, different layered cells can be observed to grow along with the outboard of the scaffold with mucosa, submucosa, muscular layer, and the serosa layer order after 7 weeks. Mucosa and muscular layer growth along the scaffold inner wall were seen simultaneously. Conclusion: 3-dimensional electrospinning synthesized 4 : 6 polycaprolactone–silk fibroin nanofiber scaffolds that are feasible for tissue growth and achieve the purpose of ureteral reconstruction in animal experiments. This new form of 3-dimensional electrospinning constructed polycaprolactone–silk fibroin nanofiber scaffold may be considered as a clinical urinary tract tissue reconstruction alternative in the future. Cite this article as: Hu H-Y, Wu C-L, Huang C-S, Bai M-Y, Yu D-S. The study of 3D printing-assisted electrospinning technology in producing tissue regeneration polymer-fibroin scaffold for ureter repair. Turk J Urol. 2022;48(2):118-129.

Background: Psychological distress is an important component of cancer survivorship, particularly among patients undergoing radical cystectomy (RC) for bladder cancer. However, the prognostic significance of postoperative psychiatric medication exposure following RC remains unclear. Aims: To investigate the association between postoperative antidepressant and/or anxiolytic exposure and survival outcomes following RC for bladder cancer. Methods: This retrospective propensity score–matched cohort study used data from the TriNetX US Collaborative Network. Adult patients who underwent RC for bladder cancer (2005–2025) were identified. Postoperative psychiatric medication exposure was defined as new use of antidepressants (Anatomical Therapeutic Chemical: N06A) and/or anxiolytics (ATC: N05B). The primary analysis evaluated combined antidepressant/anxiolytic exposure, with separate antidepressant‐only and anxiolytic‐only subgroup analyses. Overall survival (OS) and all‐cause mortality were evaluated after 1:1 propensity score matching (PSM). Results: The primary analysis identified 2461 exposed and 7257 unexposed patients, yielding 1521 matched pairs after PSM. Patients exposed to postoperative psychiatric medication were associated with significantly poorer OS than their matched controls (hazard ratio [HR]: 1.377, 95% confidence interval [CI]: 1.261–1.502; log‐rank p < 0.001). Similar associations were observed in subgroup analyses. Antidepressant‐only exposure was associated with poorer OS (HR: 1.260, 95% CI: 1.161–1.367; p < 0.001), and the anxiolytic‐only subgroup demonstrated the strongest association with poorer survival (HR: 1.457, 95% CI: 1.324–1.604; p < 0.001). Conclusions: Psychiatric medication use may represent a clinically observable marker of psychological burden and survivorship vulnerability following RC. Early recognition of postoperative psychological vulnerability may facilitate timely psycho‐oncological support and multidisciplinary survivorship care following RC.