Gemcitabine is a widely used intravesical chemotherapeutic agent for non-muscle-invasive bladder cancer (NMIBC), yet its efficacy remains suboptimal, with high recurrence rates. While traditionally considered a cytotoxic agent, emerging evidence suggests that certain chemotherapeutics may exert immunomodulatory effects. This study investigated whether gemcitabine induces immunogenic cell death (ICD) and explored the underlying mechanisms involving endoplasmic reticulum (ER) stress. A retrospective analysis was conducted to assess the association between gemcitabine-related complications and NMIBC recurrence. In vitro assays were used to evaluate ICD hallmarks (ATP, HMGB1, CALR) and pro-inflammatory cytokines. RNA sequencing and western blotting were performed to explore ER stress pathways. Functional assays included dendritic cell (DC) maturation, phagocytosis, and CD8⁺T cell proliferation and cytotoxicity. In vivo vaccination and T cell depletion models were used to evaluate antitumor immunity. Clinical data revealed an inverse correlation between gemcitabine-related complications and tumor recurrence. Gemcitabine induced classical ICD markers in bladder cancer cells and elicited CD8⁺T cell-mediated antitumor immunity in vivo. Mechanistically, gemcitabine activated the PERK branch of the ER stress response, promoting CALR translocation and DAMP release. PERK overexpression enhanced DC maturation, phagocytosis, and stimulation of CD8⁺T cell proliferation and cytotoxicity, whereas PERK knockdown impaired these effects. This study demonstrates that short-term, high-dose gemcitabine induces PERK-dependent ICD, which enhances antitumor immune responses via DC and CD8⁺T cell activation. These findings provide a mechanistic rationale for combining gemcitabine with immunotherapy and suggest PERK as a potential biomarker and therapeutic target for ICD enhancement in bladder cancer.

Conditionally replicative oncolytic adenoviruses (CRAds) display significant anti-tumor effects. However, the traditional adenovirus of serotype 5 (Ad5) entering cancer cells via coxsackie virus and adenovirus receptor (CAR) can’t be utilized for bladder cancer with low expression of CAR, which limits the application of Ad5. We utilized Ad5/F11p containing the chimeric fiber gene encoding the Ad5 fiber tail domain and Ad11p fiber shaft and knob domains to construct bladder cancer-specific chimeric type viruses Ad5/F11p-PSCAE-UPII-E1A, which can infect bladder cancer cells mediated by CD46 molecule. We carried out series of experiments in vitro to research anti-tumor effect of Ad5/F11p-PSCAE-UPII-E1A and the interaction in combination with cisplatin. The results demonstrated Ad5/F11p-PSCAE-UPII-E1A could infect bladder cancer cells (T24, EJ and 5637) in a CAR-independent way, and exert anti-tumor effect by blocking the cancer cells in G1 phase and inducing apoptosis. Ad5/F11p-PSCAE-UPII-E1A plus cisplatin enhanced the anti-proliferative effect and increased the number of apoptotic cells compared with viruses or cisplatin alone. Ad5/F11p-PSCAE-UPII-E1A plus cisplatin could upregulate the proteins expression of p53, Bax, and cleaved caspase-3, and downregulated Bcl-2 protein expression in T24, EJ and 5637 cells. We constructed a bladder cancer-specific oncolytic adenovirus and provided new combination treatment strategies for bladder cancer.

Chemotherapy remains the main treatment for muscle‐invasive bladder cancer (BLCA) despite drug resistance and lack of target drugs greatly limiting long‐term survival of patients. Thus, novel and effective drugs specific to BLCA are required to aid in its treatment and improve patient survival. In the present study, we found that the compound Ro‐31‐8220, a pan‐protein kinase C inhibitor, displays potent anti‐bladder cancer efficacy in vitro and in vivo . Ro‐31‐8220 treatment suppressed bladder cancer cell migration and invasion and also induced cell apoptosis in a dose‐dependent manner. Proteomic analysis showed that Ro‐31‐8220 treatment altered the expression of numerous proteins and KEGG enrichment analysis demonstrated that multiple signal pathways are regulated by Ro‐31‐8220, including autophagy. To further validate these results, we carried out western blotting, GFP‐LC3 fusion protein and transmission electron microscopy analyses, all of which demonstrated that Ro‐31‐8220 induced bladder cancer cell autophagy. Blockade of autophagy with chloroquine, an autophagy inhibitor, attenuated Ro‐31‐8220 induced bladder cancer cell death. In a bladder cancer xenograft tumor growth mice model, we showed that intraperitoneal injection of Ro‐31‐8220 significantly decreased tumor size and tumor weight compared to the control group, suggesting an in vivo tumor suppression ability of Ro‐31‐8220 through activation of autophagy. These results suggest that Ro‐31‐8220 may be a novel promising candidate drug for bladder cancer therapy. Further studies, including clinical trials, are required to validate these results.

Prostate cancer is the most common cancer and remains a leading cause of cancer-related deaths among men worldwide. Androgen deprivation therapy continues to be the cornerstone of treatment for prostate cancer. However, the efficacy of this treatments is often limited, leading to the emergence of drug resistance and tumor recurrence. TAK-901, an inhibitor of Aurora kinase B, has been shown to inhibit tumor growth both in vitro and in vivo models. To date, the effect of TAK-901 on prostate cancer and the underlying mechanism remain unknown. In this study, we found that TAK-901 could inhibit proliferation, colony formation and migration, while also inducing apoptosis in prostate cancer cells. We further demonstrated that TAK-901 activates the CHK1 signaling pathway, leading to G2/M-phase arrest in these cells. Additionally, we identified EPHA2 as a novel therapeutic target of TAK-901. By mutating the binding sites between EPHA2 and TAK-901, we discovered that these mutations could reverse the anti-proliferative effects of TAK-901 in prostate cancer models. Our study is the first to reveal that TAK-901 induces apoptosis in prostate cancer cells and inhibits cell growth by targeting EPHA2. These findings provide valuable insights into the underlying mechanisms of TAK-901 and may develop its therapeutic applications in prostate cancer.

Objective: Endometrial cancer (EC) remains a significant clinical challenge, particularly for patients with advanced or recurrent disease. This study aims to investigate the effects of Sanguinarine Chloride (S.C), a natural benzophenanthridine alkaloid with broad anti-tumor properties, on EC cell growth and invasion, and to elucidate its underlying molecular mechanisms. Methods: S.C’s effects on EC cell viability, proliferation, invasion, and apoptosis were evaluated using CCK-8, EdU, colony formation, 3D matrigel drop assay, FACS and Western blotting (WB). To evaluate its effects on ferroptosis, malondialdehyde (MDA) assay kits, DCFH-DA and the C11 BODIPY581/591 probe, were employed. The molecular mechanisms through which S.C regulates FTO-ACSL4 axis were investigated using plasmid transfection and WB. Additionally, a mouse xenograft model derived from EC cells was established to evaluate the in vivo effects of S.C and its molecular mechanisms, utilizing hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC) and WB. Results: S.C significantly inhibited EC cell growth and invasion. It induced cell death primarily through ferroptosis, as inhibitors (Ferrostatin-1, Deferoxamine) reversed this effect. S.C downregulated the RNA demethylase FTO, leading to increased ACSL4 expression, enhanced lipid peroxidation, suppression of the NRF2-GPX4 axis, and activated NCOA4-mediated ferritinophagy. Knocking down or pharmacologically inhibiting ACSL4 reduced S.C-induced ferroptosis. Furthermore, in a murine xenograft model, S.C significantly suppressed tumor growth, which was associated with consistent alterations in these ferroptosis-related markers in vivo. Conclusions: Our findings reveal that S.C triggers ferroptosis in EC via the novel FTO-ACSL4 axis, highlighting its potential as a therapeutic agent and identifying the FTO-ACSL4 pathway as a promising target for endometrial cancer treatment.