e15193 Background: Butyrophilin subfamily 3 member A3 (BTN3A3) is expressed in various tumors, where it modulates glycolysis, proliferation, invasion, and migration to influence progression. However, its molecular mechanisms in nasopharyngeal carcinoma (NPC) remain unclear. This study aims to elucidate BTN3A3's role in NPC progression, identify biomarkers for invasion/metastasis risk stratification, and provide foundations for novel therapeutic targets. Methods: Immunohistochemistry (IHC) profiled BTN3A3 expression in nasopharyngeal tissues and correlated it with clinicopathological parameters and prognosis. BTN3A3 was overexpressed or silenced in NPC cell lines. Proliferation and tumorigenicity were assessed via CCK-8, colony-formation, and subcutaneous xenograft assays. Cell motility was quantified by wound-healing and transwell assays. RNA-sequencing and KEGG pathway enrichment were used to identify BTN3A3-regulated pathways. Western blotting evaluated its impact on glycolytic proteins. Integrated analysis of mass-spectrometry interactome data and the GeneCards glycolysis gene set identified PKM2 as a candidate binding partner. The BTN3A3–PKM2 interaction was predicted by molecular docking and validated by co-immunoprecipitation and immunofluorescence co-localization. Functional rescue experiments were conducted using PKM2 knockdown. Results: BTN3A3 mRNA and protein levels were significantly elevated in NPC versus chronic inflammation tissues (P < 0.001), and further increased in metastatic primary tumors (P = 0.003). Multivariate Cox analysis identified BTN3A3 as an independent predictor of poorer overall and progression-free survival. KEGG enrichment of RNA-seq data identified glycolysis as the top pathway regulated by BTN3A3, and western blotting confirmed BTN3A3 manipulation altered key glycolytic enzymes (GLUT1, HK2, LDHA; P < 0.01). Integrated interactome analysis nominated PKM2 as a candidate partner, and molecular docking predicted a high-affinity BTN3A3–PKM2 interaction, which was validated by co-immunoprecipitation and co-localization. BTN3A3 increased total and phospho-PKM2-Ser37 levels. Functionally, PKM2 knockdown attenuated BTN3A3-driven increases in proliferation, invasion, migration, and glycolytic flux (P < 0.05). Furthermore, BTN3A3 promoted PKM2 nuclear translocation, and enhanced the association between ERK1/2 and PKM2, indicating it facilitates ERK1/2-mediated phosphorylation and nuclear shuttling of PKM2 to drive glycolysis and NPC progression. Conclusions: BTN3A3 is upregulated in NPC and associated with poor prognosis and distant metastasis. Functionally, it promotes glycolytic reprogramming and tumor progression by enhancing ERK1/2-dependent phosphorylation of PKM2 at Ser37 and PKM2 nuclear translocation.

PURPOSE:The efficacy and safety of TPF-induced chemotherapy(IC) combined with concurrent chemoradiotherapy(CCRT) compared to CCRT and sequential PF-adjuvant chemotherapy(AC) lack phase III randomized controlled clinical trials for evaluation, so the comparative efficacy and safety between the two approaches remain unclear. METHODS AND MATERIALS:This randomized clinical phase III trial recruited patients from May 2018 to July 2021,at 4 institutions in China(NCT03574324), 266 patients were enrolled and randomly assigned to either the IC or AC groups. The IC group received TPF followed by CCRT, while the AC group received CCRT followed by PF. We are reporting on the primary outcome of progression-free survival (PFS) and secondary endpoints of overall survival(OS), locoregional relapse-free survival(LRFS), distant metastasis-free survival(DMFS), and toxicity profile. RESULTS:The 3-year PFS was similar between the two groups, with 79 % for the IC group and 74.5 % for the AC group (P = 0.454) at a median follow-up of 39 months. Similar findings were observed, with no significant disparities in OS, LRFS, and DMFS between the two treatment cohorts. Both groups had similar compliance rates for radiotherapy and chemotherapy. However, the IC group experienced fewer grade toxic effects during CCRT, such as nausea/vomiting, swallowing, and dryness (101[77.10 %] vs. 114[89.06 %] patients,40 [30.53 %]vs56 [43.75 %] patients and 58 [44.27 %]vs86 [67.19 %] patients, respectively). However,3-4 grade leukopenia and neutrophilia patients were increased(58 [44.27 %]vs28 [21.88 %] patients and 78 [59.54 %]vs24 [18.75 %]) in the IC group. CONCLUSIONS:In this randomized clinical trial, IC did not improve 3-year PFS for LA-NPC patients and increased hematological toxicity. Still, the advantage of it is that it did reduce the incidence rates of nausea/vomiting, swallowing, and dry mouth during radiotherapy.

IRF2 is an interferon regulatory factor with context-dependent roles in cancer. We examined IRF2 expression and function in nasopharyngeal carcinoma (NPC) using immunohistochemistry, immunofluorescence, western blot, and functional assays in cell lines and a xenograft model. IRF2 was upregulated and predominantly nuclear in NPC, correlating with advanced T stage and higher EBV DNA load. ROC analysis indicated diagnostic value (AUC = 0.837). IRF2 overexpression promoted proliferation, migration, invasion, epithelial-mesenchymal transition (EMT) and suppressed apoptosis, whereas knockdown inhibited these phenotypes and reduced tumor growth in vivo. Mechanistically, IRF2 activated the Wnt/β-catenin pathway by upregulating β-catenin and facilitating its nuclear translocation. These findings establish IRF2 as an oncogenic driver in NPC and suggest the IRF2/Wnt/β-catenin axis as a potential therapeutic target.

6100 Background: To investigate whether TPF induction chemotherapy combined with concurrent chemoradiotherapy (CCRT) can provide greater survival benefits compared to CCRT followed by PF adjuvant chemotherapy. Methods: Patients with newly diagnosed locally advanced (Stage III–IVA) nasopharyngeal carcinoma (NPC) treated at the Affiliated Tumor Hospital of Guizhou Medical University, the Second Affiliated Hospital of Guizhou Medical University, the Second Affiliated Hospital of Zunyi Medical University, and Guiyang Hospital of Guizhou Aviation from May 2018 to July 2021 were enrolled. Each group included 133 patients. The experimental group received 3 cycles of TPF induction chemotherapy (docetaxel 75mg/m²,intravenous infusion, Day 1; cisplatin 75mg/m²,continuous intravenous infusion over 5 days,10:00–22:00 daily; fluorouracil 750mg/m²/day, continuous intravenous infusion over 5 days, 22:00–10:00 daily) followed by 2–3 cycles of concurrent chemotherapy (cisplatin 100mg/m², continuous intravenous infusion over 2 days, 10:00–22:00 daily). The control group received PF adjuvant chemotherapy (cisplatin 80mg/m², continuous intravenous infusion over 5 days,10:00–22:00 daily; fluorouracil 800mg/m²/day, continuous intravenous infusion over 5 days, 22:00–10:00 daily) combined with 2–3 cycles of concurrent chemotherapy (same as induction chemotherapy).Both groups underwent intensity-modulated radiotherapy (IMRT),with total doses of 69.96 Gy for T1–T2 primary lesions, 72.6 Gy for T3–T4 lesions, and 69.96 Gy for positive lymph nodes. Data were analyzed using SPSS 26.0.Differences in 5-year PFS, OS, LRFS, DMFS, and adverse events were compared between the two groups. Results: No significant differences were observed between the two groups in age, sex, Karnofsky Performance Status (KPS) score, T stage, N stage, or overall stage ( P > 0.05).At a median follow-up of 58 months, the 5-year PFS in both the intention-to-treat and per-protocol populations was similar between the induction chemotherapy (IC) and adjuvant chemotherapy (AC) groups (66.6% vs 66.0%, P = 0.589; 75.3% vs 69.9%, P =0.471). The 5-year OS, LRFS, and DMFS rates were 73.0% vs 71.3% ( P =0.582), 87.4% vs 90.8% ( P =0.508), and 76.9% vs 72.6% ( P =0.267), respectively, with no significant differences. Conclusions: Both groups had similar 5-year PFS, OS, LRFS, DMFS, and long-term toxicity profiles. Clinical trial information: NCT03574324 . 5-year observation indicators for two groups. Observation indicators IC+CCRT CCRT+AC P value Risk ratio(95% CI) PFS ITT Population 66.6% 66.0% 0.589 0.89（0.58-1.36） PP Population 75.3% 69.9% 0.471 0.85（0.55-1.33） OS 73.0% 71.3% 0.582 0.88（0.55-1.39） LRFS 87.4% 90.8% 0.508 1.30（0.60-2.80） DMFS 76.9% 72.6% 0.267 0.75（0.45-1.25） OS, LRFS, and DMFS were all calculated in the ITT population.

Radioimmunotherapy for lung cancer is effective but could cause cardiac damage. Our previous research indicated that combining radiotherapy with PD-1 inhibitors leads to myocardial injury, marked by increased HMGB1 and pyroptosis-related proteins in heart tissue. Pyroptosis, a pro-inflammatory form of programmed death, releases abundant inflammatory cytokines that further amplify tissue damage. Notably, in addition to its pivotal role in cardiac injury, HMGB1 exerts context-dependent, dual effects within the tumor microenvironment—either tumor-promoting or tumor-suppressive—whose net impact remains undefined. Therefore, this study proposes to specifically block HMGB1 in a lung cancer model to determine whether this intervention can suppress pyroptosis, alleviate cardiac damage, and to concurrently evaluate its potential impact on the tumor control, thereby providing a novel therapeutic strategy to mitigate the cardiotoxicity of radiation–immunotherapy. We established a cardiac injury model in tumor-bearing mice using radiation and PD-1 inhibitors to assess the impact of HMGB1 blockade on heart and tumor treatment. We evaluated cardiac injury and fibrosis with HE and Masson staining, assessed cardiac function via echocardiography and detect the level of cytokine in heart by ELISA. Lymphocyte infiltration was analyzed by flow cytometry, while immunofluorescence, immunohistochemistry, Western blotting, and PCR examined changes of HMGB1 and pyroptosis pathways. Additionally, we monitored tumor growth and necrosis following HMGB1 blockade. Recent research demonstrates that the combination of radiotherapy and PD-1 inhibitors significantly exacerbates myocardial injury compared to radiotherapy alone. This exacerbation is evidenced by elevated levels of inflammatory cytokines, including HMGB1, IL-1β, and IL-18, within the myocardium, along with increased fibrosis and pyroptosis. Additionally, there is an upregulation of the pyroptosis pathway, specifically the HMGB1-Caspase-1-GSDMD axis. The incorporation of an HMGB1 neutralizing antibody into the combined treatment regimen has been shown to down-regulate HMGB1 and proteins associated with pyroptosis, thereby substantially reducing cardiotoxicity and myocardial injury. Notably, the administration of the antibody does not compromise the anti-tumor efficacy of the combined regimen, as indicated by comparable tumor growth, necrosis levels, and peripheral blood lymphocyte distribution relative to the group receiving the combined treatment without the antibody. HMGB1 blockade attenuates cardiac injury caused by radiotherapy combined with PD-1 inhibitor while maintaining the anti-tumor efficacy. This demonstrates that in the context of tumor treatment, targeting HMGB1 represents a promising option for alleviating the cardiac damage caused by combined therapy.

e18052 Background: The recurrence and progression of nasopharyngeal carcinoma (NPC) are major factors contributing to unsatisfactory treatment outcomes. The role of Interferon Regulatory Factor 2 (IRF2) in NPC progression remains unclear. This study aimed to elucidate the biological functions, underlying mechanisms, and clinical significance of IRF2 in NPC. Methods: Subcellular localization and pan-cancer expression profiling of IRF2 were analyzed using the Human Protein Atlas database. Localization and expression levels of IRF2 were assessed in NPC cell lines (CNE1, HONE1, 58F, 6-10B), 64 NPC tissues, 36 chronic nasopharyngitis tissues, and the immortalized nasopharyngeal epithelial cell line NP69 via immunohistochemistry (IHC), immunofluorescence (IF) and Western blot (WB). Its diagnostic value was evaluated using receiver operating characteristic (ROC) curve analysis. Stable IRF2-overexpressing and knockdown NPC cell lines were established. Functional studies were conducted using CCK-8, colony formation, wound healing, Transwell, and flow cytometry assays, as well as a xenograft tumor model in nude mice. The effects of IRF2 on epithelial-mesenchymal transition (EMT) markers and key proteins of the Wnt/β-catenin pathway (GSK-3β, β-catenin) were examined by WB. Nuclear and cytoplasmic fractionation combined with IF was used to observe β-catenin nuclear translocation. Results: IRF2 was significantly upregulated in NPC, primarily localized in the nucleus, and demonstrated diagnostic value (AUC = 0.837, 95% CI: 0.760–0.915). Its expression level positively correlated with T stage and plasma Epstein-Barr virus (EBV) DNA load. IRF2 overexpression promoted NPC cell proliferation, migration, and invasion, while suppressing apoptosis; conversely, IRF2 knockdown produced opposite effects. In vivo experiments confirmed that IRF2 promoted tumor growth. Mechanistically, IRF2 induced EMT (downregulating E-cadherin, upregulating N-cadherin, Snail1, Snail2, ZEB1) and activated the Wnt/β-catenin signaling pathway by upregulating GSK-3β and β-catenin and promoting β-catenin nuclear translocation. Conclusions: IRF2 is primarily located in the nucleus of NPC cells and functions as an oncoprotein, whose expression is associated with local tumor invasion and plasma EBV DNA load. IRF2 drives NPC progression by promoting cell proliferation, migration, invasion, EMT, and inhibiting apoptosis, potentially through activation of the Wnt/β-catenin signaling pathway, suggesting its potential as a therapeutic target.

Abstract Radiotherapy has emerged as a cornerstone modality in the management of malignant tumors. Its evolution from a localized intervention to a radical curative strategy has been particularly pronounced in lung cancer, where its role has shifted from symptom palliation to survival prolongation. Radiation esophagitis (RE) is one of the most common acute adverse effects of thoracic radiotherapy, particularly in patients with lung cancer. In the early phase of RE, the predominant features include reactive oxygen species‐mediated DNA damage and disruption of the epithelial barrier. The inflammatory phase is characterized by nuclear factor kappa B activation, edema, and pain. During the dysregulated repair phase, impaired regeneration of the esophageal mucosal epithelium, compounded by microbiota imbalance, further exacerbates tissue injury. In the late phase, transforming growth factor beta‐driven fibrosis may culminate in stricture or fistula formation. Clinically, the condition primarily manifests as dysphagia and odynophagia. The prevention and management of RE remain significant clinical challenges. This review examines key issues related to RE within the context of precision radiotherapy, with particular emphasis on its incidence and current management strategies.

To characterize the differences of dynamic changes for absolute lymphocyte count (ALC) among esophageal squamous cell carcinoma (ESCC) patients treated with neoadjuvant chemoradiotherapy (nCRT) with or without pembrolizumab, as well as to investigate the clinical and lymphocyte-related organs dosimetric parameters that would impact ALC nadir during nCRT. A total of 216 ESCC patients who received nCRT (with pembrolizumab 144; without pembrolizumab: 72) were identified from a prospective cohort. Weekly and 1-month post-nCRT ALC were identified. lymphocyte-related organs at risk (LOARs) were delineated. linear and logistic regression analysis was used to analyze the association between G4 lymphopenia/lymphopenia nadir and clinical/DVHs factors. Receiver-operating characteristic curves were used to derive optimal dosimetric planning constraints. Grade 4 (G4) lymphopenia was defined as ALC < 0.2 × 109/L during nCRT. G4 lymphopenia was observed in 35 ESCC patients (16.2

Purpose: Analyze the incidence and risk factors of thyroid dysfunction in patients with advanced nasopharyngeal carcinoma (LA-NPC) after intensity-modulated radiotherapy (IMRT) and PD-1 inhibitor treatment and their relationship with treatment efficacy and prognosis. Methods: Eighty-five LA-NPC patients treated with IMRT and PD-1 inhibitors were retrospectively collected from March 1, 2019, to May 30, 2022. The incidence of thyroid dysfunction after combination therapy was analyzed. The Kaplan-Meier method was used to analyze the relationship between thyroid dysfunction and patient prognosis. Logistic regression analysis was used to screen independent risk factors for thyroid dysfunction. Results: As of data cutoff (May 31, 2024), the median follow-up time was 27.8 months (range: 25.6 to 32.0 months). The median time of onset of thyroid dysfunction was 8.26 months. The incidence of thyroid dysfunction is 47.06% (40/85), with clinical hypothyroidism being the main cause at an incidence rate of 28.24% (24/85) and clinical hyperthyroidism at an incidence rate of 3.53% (3/85). The incidence of grade 1 thyroid immune-related adverse events (irAEs) was 29.41% (25/85), and the incidence of grade 2 thyroid irAEs was 17.65% (15/85). Patients with thyroid dysfunction had longer overall survival, progression-free survival, and distant metastasis- free survival at both one and two years compared to patients with normal thyroid function, but the difference was not statistically significant (p > 0.05). Multivariate logistic regression analysis showed that pretreatment lactate dehydrogenase (LDH) (p = 0.079) is an independent predictor of thyroid dysfunction after radiotherapy in combination with immunotherapy for LA-NPC. Conclusion: The study found that the addition of immunotherapy increases the risk and shortens the onset time of thyroid dysfunction in LA-NPC patients treated with chemoradiotherapy. Pretreatment LDH may serve as an independent risk factor for thyroid dysfunction for LA-NPC patients.

Objective: To investigate the involvement of the circadian clock gene BMAL1 in regulating radiosensitivity in nasopharyngeal carcinoma (NPC). Methods: The regulatory role of BMAL1 in NPC radiotherapy was investigated using 44 NPC cases and the NPC cell line CNE2, which had been genetically modified to either overexpress or silence BMAL1. Infected CNE2 cells were divided into four groups, including those with BMAL1 overexpression and BMAL1 RNA interference (RNAi), as well as their respective control groups. The group responses to different doses of radiotherapy were examined. Results: The 5-year survival rates of the BMAL1 overexpression group demonstrated a significant increase compared to those of the low expression group (p < 0.05). Furthermore, the BMAL1 overexpression group exhibited a higher tumor-growth inhibition rate (p < 0.001) and apoptotic rate (p = 0.004) following radiotherapy; a decreased proportion of S-phase cells (p < 0.001) but an increased proportion of G2/M-phase cells (p = 0.001) after 24 h of 8Gy irradiation; reduced number of cell colonies (p = 0.042), and a lower survival score (p = 0.037). Conclusions: Our findings demonstrate a positive correlation between BMAL1 expression and NPC survival, suggesting that BMAL1 promotes NPC radiosensitivity.

Acquired radio-resistance is thought to be one of the main causes of recurrent metastasis after failure of nasopharyngeal carcinoma (NPC) radiotherapy, which may be related to X-ray-induced epithelial–mesenchymal transition (EMT) activation. The circadian clock gene, BMAL1, has been shown to correlate with the sensitivity of NPCs to radiotherapy, but the specific mechanism has not been reported. NPC cells were irradiated by conventional fractionation to generate radiotherapy-resistant cells. NPC cells with BMAL1 gene stabilization/overexpression and interference were obtained by lentiviral transfection. Western blotting, colony formation analysis, cell counting kit-8 assays, wound-healing tests, Transwell assays, flow cytometry, the EDU method, nuclear plasma separation experiments, HE staining, immunohistochemical staining and TUNEL staining were performed to explore the influence and molecular mechanism of the circadian clock gene, BMAL1, on NPC-acquired radio-resistance and EMT through in vitro and in vivo experiments. The results indicated that there was a gradual downregulation of BMAL1 gene protein expression during the routine dose induction of radio-resistance in NPC cells. EMT activation was present in the radiation-resistant cell line 5-8FR, and was accompanied by the significant enhancement of proliferation, migration and invasion. The BMAL1 gene significantly increased the radiosensitivity of the radiation-resistant cell line 5-8FR and reversed the acquired radio-resistance of NPCs, which was accomplished by inhibiting the TGF-β1/Smads/Snail1 axis-mediated EMT.

6027 Background: To explore the molecular mechanism of biological clock gene BMAL1 inhibiting the proliferation and metastasis of nasopharyngeal carcinoma(NPC). Methods: 1. IHC detected the expression of BMAL1 in 41 cases of nasopharyngeal chronic inflammatory tissue and 71 cases of NPCtissue. PCR and Western blot measured the expression levels of BMAL1 in NP69 in NPC cells and human immortalized nasopharyngeal epithelial cells, respectively. 2. To detect the effect of BMAL1 on the proliferation capacity of NPCcells in vitro and in vivo; Scratch healing and Transwell invasion migration experiments detected the effect of BMAL1 on the invasion and migration ability of NPCcells in vitro. Fluorescence in vivo imaging and HE staining to determine the transfer of nude mouse tail intravenous injection model. PCR and Western blot detect the effect of BMAL1 on the expression of EMT-related markers at the transcriptional and protein levels in NPC cells. 3. A series of molecular biological means such as bioinformatics technology, ChIP experiment, double luciferase reporter gene experiment, immunofluorescence colocalization experiment and Co-IP were used to verify its mechanism. Results: 1. The expression of BMAL1 in NPCtissues and cells is reduced. The expression of BMAL1 in NPC tissues was associated with the M stage of nasopharyngeal carcinoma patients (p=0.006). 2. Overexpression of BMAL1 can inhibit the proliferation, invasion and migration ability of nasopharyngeal cancer cells, while knocking down BMAL1 is the opposite. Overexpression of BMAL1 inhibits EMT of NPCcells. 3. There are 5 BMAL1 binding sites in the TGF-β1promoter region, and BMAL1 can directly bind to the TGF-β1promoter region, which can inhibit the transcriptional activity of TGF-β1. Overexpression of BMAL1 inhibits TGF-β1/Smads pathway activity. Under the induction of recombinant human TGF-β1, the proliferation ability of NPC cells was enhanced, the activity of TGF-β1/Smads signaling pathway was enhanced, and nasopharyngeal cancer cells underwent EMT, and cell invasion and migration capacity increased. After overexpression of BMAL1, TGF-β1 no longer promotes TGF-β1/Smads signaling pathway activity, EMT is inhibited, and cell invasion and migration ability is weakened. BMAL1 combined with TGF-β1 has certain diagnostic value in predicting whether nasopharyngeal carcinoma metastasis. Conclusions: BMAL1 expression is downregulated in nasopharyngeal carcinoma, and its expression level is related to the development and metastasis of nasopharyngeal carcinoma. BMAL1 can inhibit the proliferation, epithelial-mesenchymal transformation and invasion and metastasis ability of NPC cells, and its mechanism is related to the inhibition of TGF-β1/Smads signaling pathway activity. It is speculated that it may become a new prognostic marker and therapeutic target for nasopharyngeal carcinoma.

PurposeThis study aimed to evaluate 5-year outcomes and the late toxicity profile of chrono-chemotherapy with different infusion rates in patients with locally advanced nasopharyngeal carcinoma (NPC).Methods and materialsOur retrospective analysis included 70 patients with locally advanced NPC stages III and IVB (according to the 2010 American Joint Committee on Cancer staging system). Patients were treated with two cycles of induction chemotherapy (IC) before concurrent chemoradiotherapy (CCRT) at Guizhou Cancer Hospital. The IC with docetaxel, cisplatin (DDP) and fluorouracil regimen. Patients were divided into two groups during CCRT. Using a “MELODIE” multi-channel programmed pump, DDP (100 mg/m2) was administered for 12 hours from 10:00 am to 10:00 pm and repeated every 3 weeks for 2-3 cycles. DDP was administered at the peak period of 4:00 pm in the sinusoidal chrono-modulated infusion group (Arm A, n=35). The patients in Arm B received a constant rate of infusion. Both arms received radiotherapy through the same technique and dose fraction. The long-term survival and disease progression were observed.ResultsAfter a median follow-up of 82.8 months, the 5-year progression-free survival rate was 81.3% in Arm A and 79.6% in Arm B (P = 0.85). The 5-year overall survival rate was not significantly different between Arm A and Arm B (79.6% vs 85.3%, P = 0.79). The 5-year distant metastasis-free survival rate was 83.6% in Arm A and 84.6% in Arm B (P = 0.75). The 5-year local recurrence-free survival rate was 88.2% in Arm A and 85.3% in Arm B (P = 0.16). There were no late toxicities of grade 3-4 in either group. Both groups had grade 1-2 late toxicities. Dry mouth was the most common late toxic side effect, followed by hearing loss and difficulty in swallowing. There was no statistically significant difference between Arm A and Arm B in terms of side effects.ConclusionLong-term analysis confirmed that in CCRT, cisplatin administration with sinusoidal chrono-modulated infusion was not superior to the constant infusion rate in terms of long-term toxicity and prognosis.

INTRODUCTION:Metronomic capecitabine used as an adjuvant therapy improves survival in patients with locoregionally advanced nasopharyngeal carcinoma (LA-NPC). This therapeutic approach may also contribute to improving immune function, consequently enhancing overall therapeutic efficacy. AIM:We aimed to evaluate the effect of metronomic capecitabine as adjuvant therapy on immune function and survival in cases of LA-NPC. SUBJECTS AND METHODS:28 patients with LA-NPC were enrolled in the study and equally assigned to two groups of 14 each: experimental and control group. The experimental group received induction chemotherapy + concurrent chemotherapy + adjuvant chemotherapy as well as oral capecitabine at a dose of 650 mg/m² of body surface area twice daily for 1 year, with the option to discontinue in case of intolerance. The control group did not receive additional chemotherapy or targeted drugs after the induction chemotherapy + concurrent chemoradiotherapy; however, they were followed up regularly. Changes in immune function and survival were compared between the two groups. RESULTS:The median follow-up time was 43.5 months. One year after adjuvant chemotherapy, the experimental group showed higher levels of CD8 + cells, CD28 + CD8 + cells, and activated CD8 + cells compared to the control group (P < 0.05). The CD4/CD8 ratio and proportion of monocyte-derived dendritic cells were also higher in the experimental group than in the control group, but the difference was not statistically significant (P ≥ 0.05). Comparisons of 3-year overall survival, local-regional recurrence-free survival, progression-free survival, and distant metastasis-free survival between the two groups showed percentages of 92.9% vs. 78.6%, 92.9% vs. 92.9%, 78.6% vs. 71.4%, and 85.7% vs. 0.78 0.6% respectively, but these differences were not significant (P > 0 0.05 ). CONCLUSION:Metronomic capecitabine chemotherapy was observed to induce an immunomodulatory effect in LA-NPC. TRIAL REGISTRATION:NCT02958111, date of registration 04-11-2016.

Immunotherapy is currently one of the most viable therapies for head and neck squamous cell carcinoma (HNSCC), characterized by high immune cell infiltration. The Wnt-signaling inhibitor and immune activation mediator, Dickkopf-1 (DKK1), has a strong correlation with tumor growth, tumor microenvironment, and, consequently, disease prognosis. Nevertheless, it is still unclear how DKK1 expression, HNSCC prognosis, and tumor-infiltrating lymphocytes are related. To better understand these associations, we examined how DKK1 expression varies across different tumor and normal tissues. In our study, we investigated the association between DKK1 mRNA expression and clinical outcomes. Next, we assessed the link between DKK1 expression and tumor immune cell infiltration. Additionally, using immunohistochemistry, we evaluated the expression of DKK1 in 15 healthy head and neck tissue samples, and the expression of CD3, CD4, and DKK1 in 27 HNSCC samples. We also explored aberrant DKK1 expression during tumorigenesis. DKK1 expression was remarkably higher in HNSCC tissues than in healthy tissues, and was shown to be associated with tumor stage, grade, lymph node metastasis, histology, and a dismal clinical prognosis in HNSCC. DKK1 expression in HNSCC tissues was inversely correlated with CD3+ (P < 0.0001) and CD4+ (P < 0.0001) immune cell infiltration, while that in immune cells was inversely associated with HNSCC prognosis. These findings offer a bioinformatics perspective on the function of DKK1 in HNSCC immunotherapy and provide justification for clinical research on DKK1-targeted HNSCC treatments. DKK1 is a central target for improving the efficacy of HNSCC immunotherapy.

PURPOSE:This prospective clinical trial aims to compare the efficacy and safety of gemcitabine plus cisplatin (GP) versus docetaxel plus cisplatin and fluorouracil (TPF) as induction chemotherapy combined with locoregional radiotherapy in de novo metastatic nasopharyngeal carcinoma (dmNPC). METHODS:146 dmNPC patients were randomly assigned in a 1:1 ratio to receive 4-6 cycles of GP (GP group) or TPF induction chemotherapy (TPF group) followed by locoregional radiotherapy (LRRT). The primary endpoint was overall survival (OS). Secondary endpoints consisted of progression-free survival（PFS）, objective response rate (ORR), disease control rate (DCR), and treatment-related adverse events (AEs). RESULTS:As of data cutoff (May 31, 2024), the median follow-up time was 60.0 months (IQR 40.3-68.1). There is no significant difference in median OS (35.4 vs. 34.8 months, p = 0.2609) and PFS (15.8 vs. 14.3 months, p = 0.2318) between the GP and TPF groups. No significant differences in ORR (65.8 % vs. 71.2 %, p = 0.476) and DCR (79.5 % vs. 82.2 %, p = 0.674) were observed between GP and TPF group too. Furthermore, the 5-year OS was 40.1 % (95 % CI, 29.6 %-54.2 %) in the GP group, compared with 27.2 % (95 % CI, 17.9 %-41.3 %) in the TPF group（HR = 0.79, 95 % CI, 0.53-1.20). However, the TPF group had higher incidences of grade 3-4 AEs such as neutropenia, leukopenia, nausea, and diarrhea. CONCLUSION:The study indicates that 4-6 cycles of TPF induction chemotherapy combined with LRRT achieves a therapeutic effect comparable to the GP regimen with controllable safety.

Objective To understand the effects of clock gene BMAL1 and HIF-1α(Hypoxia inducible factor-1α) on proliferation, migration and sensitivity to radiotherapy of nasopharyngeal carcinoma cells HONE1.At the same time, whether the biological clock gene BMAL1 can affect the expression of HIF-1α protein was investigated.It will lay the foundation for further study on the correlation between clock gene BMAL1 and HIF pathway. Methods BMAL1 gene overexpression and interference lentivirus and HIF-1α gene interference lentivirus were constructed respectively, and were transfected into nasopharyngeal carcinoma cells HONE1. Western blot was used to verify the establishment of overexpressed and knockdown BMAL1 cell lines and HIF-1α gene knockdown cell line, and to investigate the expression of HIF-1α protein in overexpressed and knockdown BMAL1 cell lines.CCK-8 cell proliferation test and scratch test were used to analyze the proliferation and migration ability of cells.Cell apoptosis after radiotherapy was analyzed by flow cytometry.The effects of BMAL1 and HIF-1α on the sensitivity of HONE1 radiotherapy in nasopharyngeal carcinoma cells after X-ray irradiation at different doses (0Gy, 2Gy, 4Gy, 6Gy) were detected by clone formation assay. Results The overexpression of BMAL1 gene and lentivirus interference were constructed to effectively up regulate and down regulate the expression of BMAL1 protein in nasopharyngeal carcinoma cells HONE1.Meanwhile, HIF-1α gene interference lentivirus was constructed to effectively down-regulate the expression of HIF-1α protein in nasopharyngeal carcinoma cell line HONE1, and successfully screen out stable nasopharyngeal carcinoma cell lines.Western blot results showed that overexpression of BMAL1 gene could inhibit the expression of HIF-1α protein in HONE1 of nasopharyngeal carcinoma cells, while knockdown of BMAL1 gene promoted the expression of HIF-1α protein in HONE1 of nasopharyngeal carcinoma cells( P < 0.05).CCK-8 cell proliferation and scratch test showed that overexpression of BMAL1 gene or knockdown of HIF-1α gene could inhibit the proliferation and migration of HONE1 cells ( P < 0.05).Flow cytometry results showed that after 8Gy irradiation for 72 h, the apoptosis rate of BMALl gene overexpression group was higher than that of the overexpression control group, similarly, the apoptosis rate of HIF-1α gene knockdown group was higher than that of the knockdown control group ( P < 0.05).After X-ray irradiation at different doses (0Gy, 2Gy, 4Gy, 6Gy), clon-formation experiment showed that the clon-formation rate and cell survival fraction of BMALl overexpression group or HIF-1α knockdown group were lower than those of negative control group ( P < 0.05).Sigmaplot analysis showed that the D0, Dq and SF2 of the BMAL1 overexpression group or HIF-1α knockdown group were lower than those of the negative control group, and the radiosensitization ratios were 1.381 and 1.063, respectively. Conclusion Overexpression of BMAL1 gene can inhibit the proliferation and migration of nasopharyngeal carcinoma cell line HONE1, increase apoptosis after radiotherapy and improve radiosensitivity.Knock down HIF-1α Gene can inhibit the proliferation and migration of nasopharyngeal carcinoma cell line HONE1, increase apoptosis after radiotherapy and improve radiosensitivity.In nasopharyngeal carcinoma cells HONE1, overexpression of BMAL1 gene can inhibit the expression of HIF-1α protein while knockdown of BMAL1 gene can promote the expression of HIF-1α protein.

ObjectivesProthrombin time (PT) and PT-INR are independent predictors of mortality in patients with cancer. The PT and PT-INR of cancer patients are independent predictive variables of mortality. However, whether the PT or PT-INR is related to in-hospital mortality in severely ill patients with tumors remains unknown.DesignThis was a case–control study based on a multicenter public database.SettingsThis study is a secondary analysis of data extracted from 2014 to 2015 from the Electronic Intensive Care Unit Collaborative Research Database.ParticipantsThe data relevant to seriously ill patients with tumors were obtained from 208 hospitals spread throughout the USA. This research included a total of 200,859 participants. After the samples were screened for patients with combination malignancies and prolonged PT-INR or PT, the remaining 1745 and 1764 participants, respectively, were included in the final data analysis.Primary and secondary outcome measuresThe key evaluation methodology was the PT count and PT-INR, and the main outcome was the in-hospital mortality rate.ResultsAfter controlling for confounding variables, we found a curvilinear connection between PT-INR and in-hospital mortality (p < 0.001), and the inflection point was 2.5. When PT-INR was less than 2.5, an increase in PT-INR was positively associated with in-hospital mortality (OR 1.62, 95% CI 1.24 to 2.13), whereas when PT-INR was greater than 2.5, in-hospital mortality was relatively stable and higher than the baseline before the inflection point. Similarly, our study indicated that the PT exhibited a curvilinear connection with in-hospital mortality. On the left side of the inflection point (PT <22), a rise in the PT was positively linked with in-hospital mortality (OR 1.08, 95% CI 1.04 to 1.13, p < 0.001). On the right side of the inflection point, the baseline PT was above 22, and the in-hospital mortality was stable and higher than the PT count in the prior range (OR 1.01, 95% CI 0.97 to 1.04, 0.7056).ConclusionOur findings revealed that there is a curved rather than a linear link between the PT or PT-INR and in-hospital mortality in critically ill cancer patients. When these two laboratory results are below the inflection point, comprehensive therapy should be employed to reduce the count; when these two laboratory results are above the inflection point, every effort should be made to reduce the numerical value to a value below the inflection point.

BMAL1 is a core circadian clock gene that is expressed rhythmically in a variety of tumor cells and is related to cancer cell proliferation and chemoradiotherapy sensitivity. Radiotherapy plays an important role in the treatment of nasopharyngeal carcinoma (NPC). However, the rhythmicity of BMAL1 in NPC, as well as its precise role in radiotherapy, remains unclear. We assessed changes in BMAL1 expression over 48 h in NPC cells and normal nasopharyngeal epithelial cells NP69 using real-time quantitative polymerase chain reaction (RT-PCR) and western blotting (WB). Then, we induced the overexpression and knocked-down the levels of BMAL1 in NPC cells, and subsequently used Cell Counting Kit-8 assays to assess the proliferation of NPC cells. Xenograft tumour growth was used to evaluate the effect of BMAL1 in vivo. Immunohistochemical staining was used to detect the expression of BMAL1 protein in transplanted tumors. Gene Set Enrichment Analysis (GSEA) was performed to explore the biological signaling pathway. Finally, RT-PCR and WB were used to detect the expressions of BMAL1, p53 and p21. The results showed that the mRNA expression levels of circadian clock gene BMAL1 fluctuated rhythmically with time, and the expression levels of BMAL1 also changed depending on the protein levels in NPC and NP69 cells. Overexpression of BMAL1 inhibited the proliferation of NPC cells, while knockdown BMAL1 had the opposite effects. In a xenograft model, we observed that the upregulation of BMAL1 inhibited tumor growth and enhanced the sensitivity of NPC cells to radiotherapy. Ultimately, the downregulation of BMAL1 promoted tumor growth and decreased radiosensitivity. GSEA analysis suggested that BMAL1 significantly affected the p53 pathway. Overexpression of BMAL1 promoted the expression of p53 and p21, while the knockdown of BMAL1 inhibited the expression of p53 and p21. We speculate that BMAL1 has the potential to be a prognostic biomarker and therapeutic target for NPC.

Objective:To explore the potential mechanism of PD-1 inhibitor P on RIMI from the perspective of immune microenvironment.Methods:To establish a mouse model of radiation-induced myocardial injury (RIMI), twenty C57BL/6 mice were randomly divided into 4 groups, 5 in each group. Group A was the healthy control group; Group B was the PD-1 inhibitor group; Group C was the simple irradiation group, with a heart irradiation of 15 Gy; Group D was the irradiation+ PD-1 inhibitor group. One month after irradiation, the mice were anesthetized and sacrificed. The morphological changes of myocardial tissues were observed by HE staining. The myocardial fibrosis was assessed by Masson staining. CD 3+ , CD 3+ CD 4+ , CD 3+ CD 8 lymphocyte subsets and cytokines (IL-4, IL-6, IL-17A, TNF-α, TGF-β 1 and INF-γ) levels were determined by flow cytometry. The apoptosis rate of myocardial cells was detected by TUNE. Results:One month after irradiation, there was no obvious myocardial fibrosis in group B, and collagen fibers were distributed in the interstitium of myocardial cells in groups C and D. Semi-quantitative analysis results showed that the myocardial collagen volume fraction (CVF) of groups A, B, C and D were (1.97±0.36)%, (2.83±1.03)%, (5.39±0.77)% and (7.72±1.43)%, respectively. The CVF between group A and group B was similar ( P=0.314), and the differences in CVF between the other groups were statistically significant (all P<0.05). Compared with group A, the absolute value and percentage of CD 3+ T lymphocytes were significantly increased in groups B, C and D (all P<0.01). The values in group D were significantly higher than those in group B and group C (all P<0.01); The absolute value and percentage of CD 3+ CD4 T lymphocytes were similar among four groups (all P>0.05); The absolute value and percentage of CD 3+ CD 8 T lymphocytes in group D were significantly higher than those in groups A, B and C (all P<0.001). The expression levels of IL-6, IL-17A, and TGF-β 1 in group D were significantly higher compared with those in groups A, B and C (all P<0.001). The apoptotic index was gradually increased in four groups, and the differences in apoptotic index among four groups were statistically significant (all P<0.001). Conclusion:PD-1 inhibitors can aggravate RIMI by promoting myocardial immune inflammatory response.