Radiation-induced lung fibrosis (RILF) is a severe and life-threatening complication of thoracic radiotherapy. Cell death is the key issue in RILF. Ferroptosis is a form programmed cell death implicated in the pathologies of inflammation. This study aimed to investigate the role of ferroptosis in RILF, and the effectiveness and the potential underlying mechanism of ferroptosis inhibitor on RILF. Immunofluorescence, western blot and RT-PCR assays were performed to examine the ferroptosis maker glutathione peroxidase 4 (GPX4) in a mice RILF model. The lung tissue sections were stained with hematoxylin and eosin (H&E), Masson trichrome staining and Sirius-Red staining to evaluate the histopathological changes in RILF mice. Reactive oxygen species (ROS) and hydroxyproline (HYP) in lungs were measured by the relevant kits. The serum levels of inflammatory cytokines (TNF-α, IL-6, IL-10, and TGF-β1) were measured with Elisa. The protein and mRNA levels of GPX4, nuclear factor (erythroid-derived 2)-like 2 (Nrf2), hemeoxygenase-1 (HO1) and quinone oxidoreductase 1 (NQO1) in lungs were examined by western blot and RT-PCR. GPX4 levels of the irradiated lungs were significantly down-regulated than the groups with no irradiation, and the ferroptosis inhibitor, liproxstatin-1, increased GPX4 levels significantly in RILF mice. Treatment with liproxstatin-1 lowered the Szapiel and Ashcroft scores significantly, down-regulated the levels of ROS and HYP in lungs and reduced the serum inflammatory cytokines levels in RILF mice. The protein and the mRNA levels of Nrf2, HO1 and NQO1 were up-regulated by liproxsratin-1 in RILF. Our data suggested that ferroptosis played a critical role in RILF, ferroptosis inhibitor liproxstatin-1 alleviated RILF via down-regulation of TGF-β1 by the activation of Nrf2 pathway. The effectiveness of ferroptosis inhibition on RILF provides a novel therapeutic target for RILF.

Radiation‐induced lung injury (RILI) is one of the most common, serious, and dose‐limiting toxicities of thoracic radiotherapy. A primary cause for this is the radiation‐induced cell death. Ferroptosis is a recently recognized form of regulated cell death, characterized by the accumulation of lipid peroxidation products and lethal reactive oxygen species (ROS). The ROS generated by irradiation might be the original trigger of ferroptosis in RILI. In addition, activation of the P62‐Kelch‐like ECH‐associated protein 1 (Keap1)‐nuclear factor erythroid 2‐related factor 2 (NRF2) pathway has been shown to blunt ferroptosis and thus acts as a protective factor. Therefore, this study aimed to explore the protective effect of the P62‐Keap1‐NRF2 pathway against radiation‐induced ferroptosis in alveolar epithelial cells. First, we found that radiation induced ferroptosis in vitro using a RILI cell model, which could be significantly reduced by ferrostatin‐1 (Fer‐1), a specific ferroptosis inhibitor. Additionally, overexpression of P62 interacted with Keap1 to facilitate the translocation of NRF2 into the nucleus and promote the expression of its target proteins, including quinone oxidoreductase 1 (NQO1), heme oxygenase 1 (HO1), and ferritin heavy chain 1 (FTH1). In summary, our results demonstrated that the activation of the P62‐Keap1‐NRF2 pathway prevents radiation‐induced ferroptosis in RILI cells, providing a theoretical basis of finding a potential therapeutic approach for RILI.

Background: The relationship of inflammation and tumor is becoming more and more important in the study on the pathogenesis of colorectal cancer. The role of TLR9-mediated immune inflammation reaction in the process is not currently clear. The purpose of the study was to discuss the correlation of TLR9 signal activation with tumor progression by detecting the expression of TLR9 and its downstream molecule NF-kappaB in colorectal cancer tissues at different stages. Methods: TLR9 expression in colorectal cancer tissues was detected by immunohistochemical streptavidin-perosidase method and Western blot. Results: The result showed that the high expression of TLR9 was correlated with tumor poorly differentiation, invasion and liver metastasis, the abnomal increasing levels of CEA in blood. With the signal activation, the levels of TLR9 protein raised more in advanced colorectal cancer than in early colorectal cancer. Afterward, we found that the activation of specific expression of TLR9 signal was related to histologic origin. TLR9-C expression displayed in both advanced cancer and para-carcinoma tissues, and TLR9-R protein was predominat in partial sigmoid and rectal cancer tissues. With the differential expression of TLR9, the levels of its downstream molecule NF-kappaB protein increased in colon cancer tissues and decreased in rectal cancer tissues. Conclusion: The results confirmed that TLR9 signaling activation participated in the clinical process of colorectal cancer and influenced NF-kappaB expression.

The radiation-induced lung injury is a common complication from radiotherapy in lung cancer. CpG ODN is TLR9 activator with potential immune modulatory effects and sensitization of radiotherapy in lung cancer. This study aimed to examine the effect of CpG ODN on acute radiation-induced lung injury in mice. The mouse model of radiation-induced lung injury was established by a single dose of 20 Gy X-rays exposure to the left lung. The results showed that the pneumonia score was lower in RT+CpG group than in RT group on 15th and 30th days. Compared with RT group, CpG ODN reduced the serum concentrations of MDA (P < 0.05) and increased the serum concentrations of SOD, GSH (P < 0.05). The serum concentration of TNF-α in RT+CpG group was lower on 15th and 30th days post-irradiation (P < 0.05). The study demonstrated that CpG ODN has preventive effects of acute radiation-induced lung injury in mice. Lung inflammatory reaction and oxidative stress are promoted in the initiation of radiation-induced pneumonia. CpG ODN may reduce the injury of reactive oxygen species and adjust the serum TNF-α concentration in the mice after irradiation, which reduces the generation of the inflammatory cytokines.

Background: Many studies have reported that the inflammatory immune response related to TLR9 signaling activation participates in tumor development and affects the treatment outcome. RUNX3 functions as a tumor suppressor by regulating DNA methylation. RUNX3 protein plays an important role in TGF-β signaling pathway that is involved in tumor growth inhibition and apoptosis. At present, radiotherapy is still an important treatment in lung cancer, which induces immune response and affects the therapeutic outcome. The role of TLR9 signaling activation and RUNX3 in this process is not clear. Methods: In this study, we investigated the expression of TLR9 in tumor and RUNX3 in surrounding tissues by immunohistochemical methods and analyzed the relationship on postoperative survival in lung cancer. Results: We found that the high expression of TLR9 was the risk factor in postoperative survival of lung cancer with no difference in lifetime. The high expression of RUNX3 in lung cancer with TLR9 signaling activation was in favor of progression-free survival and overall survival in postoperative radiotherapy. It suggested that RUNX3 played an important role in lung cancer radiotherapy. In order to determine the effect of RUNX3 in lung cancer radiation with TLR9 signaling activation, we introduced 5-Aza-2ʹ-deoxycytidine (5-Aza-CdR) and exposed lung cancer A459 cells repeatedly. The high expression of RUNX3 especially RUNX3-B in cells treated with 5-Aza-CdR was observed. We examined that 5-Aza-CdR induced more cell blocking in G2/M phase in combining irradiation. Conclusion: The result implied that it was feasible to improve radiosensitivity of lung cancer with TLR9 signaling activation by increasing RUNX3 expression, and 5-Aza-CdR was an option in this process.

Introduction: Platinum-based drugs, including cisplatin, carboplatin, and oxaliplatin, are widely used in the treatment of various malignancies. However, the route-stratified adverse-event reporting patterns have not been fully characterized. This study aims to describe and characterize the patterns and time-to-onset distributions of adverse events (AEs) for platinum drugs across intravenous (IV), intraperitoneal (IP), and intraarterial (IA) routes. Methods: Data covering the period from 2004Q1 to 2024Q4 were collected from the Food and Drug Administration Adverse Event Reporting System (FAERS) database. Disproportionality analyses were conducted to identify potential safety signals by platinum agent and route. The identified signals were then classified according to clinical priority, a categorization based on clinical relevance, reporting rate, fatality rate, and signal stability. Furthermore, time-to-onset profiles were analyzed using Kaplan-Meier analysis, the log-rank test, and Weibull shape parameter modeling. Results: The study identified AE reports associated with cisplatin (5,783 IV, 193 IP, 135 IA), carboplatin (17,091 IV, 115 IP, 171 IA), and oxaliplatin (18,845 IV, 170 IP, 108 IA). IP reports were more frequently submitted for female patients, whereas IA reports were more frequently submitted for male patients, likely reflecting route-associated clinical indications. AE reporting profiles differed by platinum agent and route, with partial overlap. Among the identified AEs, twelve AEs were classified as high clinical priority. Of these, hepatic failure was the most frequently reported high-priority AE for both IV cisplatin and IV oxaliplatin. Notably, all IV platinum agents exhibited early failure- type patterns. The cumulative reporting distributions following IV administration differed significantly by drug. Patients treated with cisplatin showed shorter time to onset, with a median of 16 days (IQR: 7-46 days). Discussion: Through a comprehensive analysis of FAERS data, this study has identified and characterized the AE signals for platinum drugs across IV, IP, and IA routes, assessed clinical priorities, and elucidated temporal characteristics. These findings should be interpreted as exploratory pharmacovigilance signals because FAERS does not support incidence estimation, comparative risk assessment, or causal inference. Therefore, large-scale prospective studies are warranted to validate and extend these results. conclusion: The distinct, drug- and route-specific AE profiles and timing patterns of platinum-based agents underscore the need for ongoing pharmacovigilance and integrated safety monitoring to enable timely risk identification and mitigation. Conclusion: This study identified differences in AE reporting profiles and time-to-onset patterns stratified by platinum agent and administration route. This underscores the importance of pharmacovigilance and the integration of safety monitoring systems to facilitate timely identification and management of AEs in clinical practice.