Unlike other types of cancers, direct implantation metastasis is a major cause of death in ovarian cancer, whereas stemness-related spheroid formation is a critical step for cancer cell colonization to the mesothelium. Despite extensive research, directly investigating ovarian cancer stemness under stem-cell-selective culture conditions remains uncommon. Resistance to the PARP inhibitor olaparib remains a major clinical challenge and is closely linked to cancer stemness. In this study, we found that Cleavage and Polyadenylation Specificity Factor 4 (CPSF4), a key regulator of pre-mRNA processing, is significantly upregulated in three-dimensional (3D)-cultured ovarian cancer spheroids and closely associated with cancer stemness. The role of CPSF4 in ovarian cancer remains unclear. Here, we demonstrate that CPSF4 contributes to drug resistance in 3D-cultured ovarian cancer cells. Mechanistically, CPSF4 upregulates CACNA2D2, a key modulator of calcium channel activity, thereby maintaining intracellular calcium homeostasis and promoting stemness-associated adaptation to therapeutic stress. Notably, CPSF4 depletion enhances sensitivity to olaparib. Loss of CPSF4/CACNA2D2 significantly sensitizes ovarian cancer cells to olaparib treatment. Collectively, these findings identify a novel CPSF4–CACNA2D2 axis in ovarian cancer and suggest a potential strategy for overcoming reduced sensitivity to olaparib.

IntroductionLung adenocarcinoma (LUAD) is the leading cause of cancer-related mortality worldwide, but its therapeutic efficacy remains suboptimal. This study explores the functional role and underlying mechanism of BUB1 in LUAD.MethodsIn vitro, BUB1 knockdown (si-BUB1) in A549/H1299 cells was performed, and effects were assessed. The ferroptosis inhibitor Fer-1 was used. Mechanistically, the role of the STAT3/GPX4 axis was investigated through overexpression experiments. In vivo, xenograft models were used.ResultsBioinformatics analysis highlighted a significant upregulation of BUB1 in LUAD tissues, with elevated expression levels correlated with reduced disease-free survival (DFS) and overall survival (OS). BUB1 knockdown markedly suppressed cell proliferation, migration, and invasion, while concurrently inducing ferroptosis. This was evidenced by typical mitochondrial morphological changes (shrinkage, increased membrane density, reduced cristae), altered ferroptosis-related markers (decreased FTH1/SLC7A11, increased COX2), elevated Fe²+/MDA levels and reduced GSH activity, which could be reversed by Fer-1. BUB1 silencing suppressed the expression and phosphorylation of STAT3, thereby downregulating the transcription of GPX4. Overexpression of STAT3 and GPX4 partially reversed the inhibitory effects of BUB1 knockdown on LUAD cell malignancy and abrogated the ferroptosis induced by BUB1 silencing. In vivo, xenograft models further validated that BUB1 silencing significantly reduces tumor volume, accompanied by modulation of ferroptosis-related genes in tumor tissues.DiscussionCollectively, our findings identify BUB1 as a novel prognostic biomarker and therapeutic target for LUAD, revealing a new regulatory mechanism by which BUB1 promotes LUAD progression through the activation of the STAT3/GPX4 axis to suppress ferroptosis.

The lack of effective therapeutic options available for microsatellite stable (MSS) colorectal cancer (CRC) remains a significant clinical challenge. Interestingly, chemotherapy-resistant cancer cells can be induced to undergo ferroptosis, prompting our investigation into RSL3, a potent ferroptosis inducer, in MSS CRC cells. Our findings revealed that while RSL3 suppressed the growth of MSS CRC cells, a subset displayed resistance. Single-cell sequencing uncovered an aberrant activation of hypoxia pathways in RSL3-resistant MSS CRC cells. Inhibiting HIF-1α, the key transcription factor driving hypoxia signaling, restored RSL3 sensitivity in these resistant cells; moreover, this sensitivity was attenuated upon HIF-1α overexpression. Chromatin immunoprecipitation assays further demonstrated that in RSL3-resistant cells, HIF-1α was enriched at the promoter of P4HA1, a gene implicated in ferroptosis resistance, thereby enhancing its expression. Additionally, in vivo experiments using syngeneic transplantation of CT26 cells in mice revealed that combining RSL3 with an HIF-1α inhibitor markedly enhanced tumor suppression and metastasis prevention, concomitant with increased intratumoral infiltration of CD8+ T cells and CD86+ macrophages. Notably, the combination enhanced the antitumor response of anti-PD1, a treatment otherwise ineffective on this tumor. These findings suggest that targeting HIF-1α represents a promising therapeutic strategy when used in conjunction with a ferroptosis inducer for the treatment of MSS CRC.

The immunosuppressive tumor microenvironment (TME) critically undermines the efficacy of T cell-based tumor immunotherapy by impeding CD8+ T cell infiltration and cytotoxic function, primarily through tumor-associated macrophages (TAMs) and immune checkpoint molecules such as programmed death ligand 1 (PD-L1). Here, we present a multifunctional nanoplatform, IN@OMV-PDL1nb, designed to simultaneously inhibit TAM-derived immunosuppressive metabolite itaconic acid (ITA) by targeting immune-responsive gene 1 (IRG1) and block PD-L1 within the TME. Engineered outer membrane vesicles (OMVs) serve as precision delivery vehicles for the IRG1 inhibitor IRG1-IN-1 (IN) and as carriers for PD-L1 nanobody release, activated by matrix metalloproteinase-2 (MMP-2). IN@OMV-PDL1nb effectively inhibits IRG1 expression in TAMs, thus reducing the accumulation of ITA, restoring chemokines (CXCL9 and CXCL10) secretion, and enhancing CD8+ T cells infiltration within tumors. The released PD-L1 nanobody protects CD8+ T cells, preserving their tumoricidal activity. In murine tumor models, IN@OMV-PDL1nb significantly inhibited tumor growth, increased survival, and enhanced antigen presentation and T cell recruitment. Additionally, IN@OMV-PDL1nb induced robust adaptive immunity, facilitating antigen-specific immune memory that prevented tumor recurrence and metastasis. This dual-targeting approach offers a promising strategy to overcome TME-driven immunosuppression in tumor immunotherapy.

Renal cell carcinoma (RCC), particularly the clear-cell subtype (ccRCC), accounts for 75-85% of kidney cancers and exhibits distinct genetic and biological heterogeneity. While surgical resection remains the mainstay of treatment for localized ccRCC, the persistence of recurrence rates underscores the significant unmet need for effective adjuvant therapies. Recent advancements in immunotherapy and targeted therapies have revolutionized the management of RCC. Immune checkpoint inhibitors have significantly enhanced antitumor immune responses, whereas tyrosine kinase inhibitors (TKIs) and mammalian target of rapamycin (mTOR) inhibitors effectively disrupt angiogenesis and proliferation signaling pathways, respectively. However, non-clear cell RCC subtypes remain understudied due to their rarity and exclusion from major clinical trials. Consequently, this review primarily focuses on ccRCC, aiming to provide a comprehensive and up-to-date overview of the latest advancements in immunotherapy and targeted therapies. By synthesizing current evidence, this review seeks to elucidate the mechanisms of action, clinical efficacy, and limitations of these treatments, while also identifying gaps in knowledge and future research directions. Ultimately, the goal is to offer valuable insights for clinicians and researchers, facilitating the development of optimized, personalized treatment approaches to improve outcomes for ccRCC patients.

V-domain immunoglobulin suppressor of T-cell activation (VISTA) is a promising next-generation immune checkpoint target. This study investigated the distribution and clinical significance of VISTA expression in cervical carcinoma. Using a cohort of 290 patients from Sun Yat-sen Memorial Hospital, we assessed VISTA expression in tumor cells, endothelial cells, and immune cells (ICs) through immunohistochemistry, and found that it was expressed in tumor cells (18.6%), endothelial cells (38.3%), and ICs (100%). Higher infiltration of VISTA+ ICs was observed in the stromal region compared with that in the intratumoral region of resectable cervical carcinoma. Among the ICs, macrophages showed the highest VISTA expression compared with that by T cells and neutrophils. High intratumoral VISTA expression was an independent marker of favorable prognosis. Robust infiltration of VISTA+ ICs and CD8+ T cells in the tumor microenvironment correlated with the best clinical outcomes. In addition, in an independent cervical carcinoma cohort (n = 48), VISTA+ ICs in both intratumoral and stromal regions were positively associated with CD8+ T cells, CD103+ T cells, and effector molecules, such as granzyme B. Elevated VISTA expression was also associated with an enriched immunosuppressive profile, including Foxp3+ regulatory T cells and molecules such as TGF-β1, PD-1, LAG-3, TIM-3, and TIGIT. Moreover, multiplex staining and correlation analysis revealed a positive association between VISTA+ TGF-β1+ ICs and FOXP3+ regulatory T cells in tumor tissues. These findings establish a strong connection between intratumoral VISTA+ ICs and a regulatory immune contexture involving both activation and suppression signatures, with a skew toward activation dominance. Our study results suggest that VISTA can be employed as a potential prognostic biomarker for cervical carcinoma.

BACKGROUND:Tumour-infiltrating lymphocytes (TILs) are strongly correlated with the prognosis and immunotherapy response in bladder cancer. The TIL status is typically assessed through microscopy as part of tissue pathology. Here, the authors developed Rad-TIL model, a novel radiomics model, to predict TIL status in patients with bladder cancer. MATERIAL AND METHODS:The authors enrolled 1089 patients with bladder cancer and developed the Rad-TIL model by using a machine-learning method based on computed tomography (CT) images. The authors applied a radiogenomics cohort to reveal the key pathways underlying the Rad-TIL model. Finally, the authors used an independent treatment cohort to evaluate the predictive efficacy of the Rad-TIL model for Bacillus Calmette-Guérin (BCG) immunotherapy. RESULTS:The authors developed the Rad-TIL model by integrating tumoral and peritumoral features on CT images and obtained areas under the receiver operating characteristic curves of 0.844 and 0.816 in the internal and external validation cohorts, respectively. Patients were stratified into two groups based on the predicted radiomics score of TILs (RSTIL). RSTIL exhibited prognostic significance for both overall and cancer-specific survival in each cohort (hazard ratios: 2.27-3.15, all P<0.05). Radiogenomics analysis revealed a significant association of RSTIL with immunoregulatory pathways and immune checkpoint molecules (all P<0.05). Notably, BCG immunotherapy response rates were significantly higher in high-RSTIL patients than in low-RSTIL patients (P=0.007). CONCLUSION:The Rad-TIL model, a noninvasive method for assessing TIL status, can predict clinical outcomes and BCG immunotherapy response in patients with bladder cancer.

Extracellular vesicles (EVs) enable the transmission of crucial molecular components between the parental and recipient cells. Macrophages can polarize into two distinct macrophage phenotypes, thereby exerting diverse effects on recipient cells. Here, we present a protocol for the direct isolation of EVs from bone marrow-derived macrophages (BMDMs) using ultracentrifugation. We describe steps for culturing BMDMs and macrophage polarization. We then detail procedures for further characterizing the isolated EVs using transmission electron microscopy (TEM), western blotting, and nanoparticle tracking analysis (NTA).

Introduction:The development of immune-related adverse events (irAEs) has been associated with improved survival from various solid tumors. Given that immunotherapy has not been widely used in ovarian cancer and has only been applied to patients with high tumor mutational burden or microsatellite instability, studies exploring the effects of irAEs on ovarian cancer have been limited. Case Presentation:A 47-year-old woman was diagnosed with International Federation of Gynecology and Obstetrics stage III ovarian cancer in 2013. Between 2013 and 2021, she underwent palliative chemotherapy comprising paclitaxel liposomes, cisplatin/nedaplatin, S-1/raltitrexed, irinotecan, doxorubicin, vinorebine, toripalimab, apatinib, gemcitabine, oxaliplatin, and capecitabine, as well as two debulking surgeries. From November 2021, she received six cycles of tislelizumab (a PD-1 inhibitor), paclitaxel (albumin-bound), and carboplatin, to which a partial response was observed according to the Response Evaluation Criteria in Solid Tumors. From May 2022, the patient was switched to maintenance therapy with tislelizumab plus olaparib. However, all antitumor treatments were discontinued from April 2023 due to multiple irAEs, including hypothyroidism, adrenal insufficiency, and pneumonitis, with the tumor remaining stable until November 2023. Progression-free survival (PFS) was approximately 24 months with tislelizumab-containing therapy but was 18 months with tislelizumab/olaparib maintenance therapy. Conclusions:We report a case involving a patient with highly pretreated recurrent ovarian cancer who exhibited prolonged PFS after developing three irAEs. The distinctly prolonged PFS observed, along with the reviewed literature, suggests that irAEs may be correlated with improved survival in ovarian cancer.

Cancer stem cells (CSCs) represent a small subset of tumor cells populations characterized by their ability to self-renew and differentiate. These cells are often considered resistant to chemotherapy, radiotherapy, and immunotherapy, playing a crucial role in driving tumor progression and metastasis. To evade immune attacks, CSCs utilize various genetic and epigenetic strategies that diminish immune recognition, enhance tolerance to immune-induced cytotoxicity, and foster the development of a protective immunosuppressive microenvironment. This microenvironment is shaped by a group of key immunosuppressive cells, particularly myeloid-derived suppressor cells (MDSCs), which not only directly inhibit effector T cells and natural killer (NK) cells, facilitating the immune escape of CSCs, but also significantly contribute to the maintenance of tumor cell stemness and promote their metastasis. Conversely, the developmental signals of MDSCs are also regulated by CSCs. This complex interplay between MDSCs and CSCs adds layers of complexity to the cancer-immune cycle and the associated tumor treatment strategies. Therefore, understanding the detrimental interdependence between MDSCs and CSCs to effectively impede tumor progression has become heated topic in tumor immunology. In this review, we provide a timely summary of the latest studies on the reported characteristics of CSCs and MDSCs, discuss their interconnection during tumor progression, and evaluate various immunotherapeutic strategies targeting these cell populations.

Cancer immune evasion is one of the important mechanisms for cancer development, which is essential to developing novel immunotherapeutic strategies. The SOX (SRY-related HMG-box) family of transcription factors plays a crucial role in normal physiology as well as in a variety of human diseases especially cancer. It has been shown that SOX is involved in cancer immune evasion processes. This mini-review aimed to summarize how SOX family members induce cancer immune evasion by regulating antigen presentation, shaping the tumor immunosuppressive milieu, and controlling regulatory immune checkpoint inhibitors like programmed death ligand 1. Thorough exploration of SOX family will help uncover the mechanism of cancer immune evasion, and provide new ideas and targets for the development of immunotherapy strategies.

Tumor-associated macrophages (TAMs) have multiple potent functions in cancer representing important therapeutic targets. MS4A4A is a functional TAM marker with controversial implications for prognosis. This study aimed to evaluate the association between MS4A4A+ TAM infiltration and clinical outcomes in urothelial carcinoma of the bladder (UCB), as well as their impact on the immune landscape. A total of 400 UCB patients from cohorts at Sun Yat-sen Memorial Hospital were analyzed. Immunohistochemistry was used to quantify MS4A4A+ TAMs and assess their spatial distribution alongside various immune components, evaluate the benefit of Bacillus Calmette-Guérin (BCG) immunotherapy, and analyze survival outcomes. Additionally, matched UCB tissues were examined before and after recurrence or progression. We observed that MS4A4A+ TAMs were present at higher levels in the stromal region compared to the intratumoral region, and correlated with advanced tumor stage and poor prognosis in both regions. No significant difference was observed in the number of MS4A4A+ TAMs before and after recurrence/progression in the same patient. Stromal MS4A4A+ TAMs were negatively correlated with BCG efficacy and recurrence-free survival. These TAMs were positively associated with CD8+ T cells, Foxp3+ regulatory T cells, immune checkpoints (PD-1, LAG-3, HAVcr-2, TIGIT), and anti-inflammatory molecules (TGF-β1, IL-4) in the same respective regions. Additionally, MS4A4A+ TAMs expressed high levels of TGF-β1 and HAVcr-2 in UCB tissues. In vitro, IL-4 induced MS4A4A expression in mouse bone marrow-derived macrophages, while Ms4a4a knockdown reduced the anti-inflammatory molecule Arg1 and increased pro-inflammatory molecule Nos2 expression. These findings demonstrate that MS4A4A is a reliable prognostic marker and predictor of BCG response in UCB, highlighting its role in shaping the immune landscape and immunotherapy outcomes. © 2025 The Pathological Society of Great Britain and Ireland.

As a crucial medical imaging modality, ultrasonography has emerged as a pivotal tool for tumor diagnosis and treatment owing to its non-invasive nature, real-time imaging capability, and superior resolution. Recent technological advancements have demonstrated unique advantages in early tumor screening, staging, and localization. Contrast-enhanced ultrasound (CEUS), utilizing microbubbles (MBs) and nanobubbles (NBs) to target vascular biomarkers, significantly enhances tumor visualization and demonstrates high sensitivity in molecular imaging. Multimodal ultrasound (MU), incorporating techniques such as elastography and automated breast volume scanning (ABVS), achieves improved diagnostic accuracy when combined with MRI/CT. The applications of ultrasound in localized and systemic tumor therapy have expanded considerably. High-intensity focused ultrasound (HIFU) enables thermal ablation of solid tumors, while low-intensity focused ultrasound (LIFU) facilitates sonodynamic therapy (SDT) through reactive oxygen species (ROS) generation mediated by sonosensitizers. Ultrasound-assisted drug delivery systems (US-DDS) leverage MB/NB cavitation effects to enhance chemotherapeutic agent delivery efficiency, overcome biological barriers, including the blood-brain barrier, and modulate immune responses. These technological breakthroughs have provided novel therapeutic options for cancer patients, garnering significant clinical interest. This review systematically examines current applications of ultrasound imaging and therapy in oncology, evaluates its potential clinical value, analyzes existing technical limitations, and discusses future development prospects. The article aims to provide innovative perspectives for tumor diagnosis and treatment while offering references for clinical practice.

Targeting cancer cell plasticity through chromatin organization is an emerging research area, yet the molecular mechanisms that govern chromatin loop formation remain unclear. Here, we develop a CRISPR screen based on our engineered live-cell CTCF-cohesin contact reporters to identify regulators of chromatin loops. Our findings reveal that tousled-like kinase 2 (TLK2) functions as a key regulator of chromatin loop formation during the cancer stemness transition. Mechanistically, TLK2 phosphorylates DYNLL1, enhancing its interaction with CTCF to promote CTCF-cohesin hub formation at the KLF4 locus. Suppressing TLK2 impairs cancer stemness plasticity, sensitizes cancer cells to cytotoxic stress in vitro, and reduces lung metastases and enhances immunotherapy response in breast cancer mouse models. Clinically, elevated TLK2 expression correlates with poor prognosis in breast cancer patients. Collectively, these findings identify TLK2 as a potential therapeutic target for mitigating cancer stemness plasticity, highlighting chromatin loop-targeting therapy as a promising strategy to eradicate cancer stem cells.

AbstractObjectivesLymphatic metastasis, an early stage of the metastasis process, is associated with adverse clinical outcomes in urothelial carcinoma of the bladder (UCB). However, the role of inflammation in triggering lymphatic metastasis remains unclear.MethodsWe employed an RNA‐sequencing cohort (n = 50) from Sun Yat‐Sen Memorial Hospital (SYMH) to identify the most highly upregulated inflammatory gene associated with lymphatic metastasis. Using immunohistochemistry and immunofluorescence analyses, we validated the association of the identified molecule with clinical features and prognosis in an independent UCB cohort (n = 244) from SYMH. We also analysed TCGA‐BLCA cohort (n = 408) to identify its potential biological pathways and immune landscape.ResultsIn our study, chitinase 3‐like 1 (CHI3L1) emerged as a significantly overexpressed proinflammatory mediator in UCB tissues with lymphatic metastasis compared to those without lymphatic metastasis (81.1% vs. 47.8%, P < 0.001). Within UCB tissues, CHI3L1 was expressed in both stromal cells (52.8%) and tumor cells (7.3%). Moreover, CHI3L1+ stromal cells, but not tumor cells, exhibited independent prognostic significance for both overall survival (P < 0.001) and recurrence‐free survival (P = 0.006). CHI3L1+ stromal cells were positively associated with D2‐40+ lymphatic vessel density (P < 0.001) and the immunosuppressive PD‐L1/PD‐1/CD8 axis in UCB tissues (all P < 0.05). A bioinformatics analysis also identified a positive association between CHI3L1 expression and lymphangiogenesis or immunosuppression pathways.ConclusionOur study established a clear association between stromal CHI3L1 expression and lymphatic metastasis, suggesting that stromal CHI3L1 expression is a potential prognostic marker for bladder cancer patients.

Long noncoding RNA-H19 ( H19 ), an imprinted oncofetal gene, has a central role in carcinogenesis. Hitherto, the mechanism by which H19 regulates cancer stem cells, remains elusive. Here we show that breast cancer stem cells (BCSCs) express high levels of H19 , and ectopic overexpression of H19 significantly promotes breast cancer cell clonogenicity, migration and mammosphere-forming ability. Conversely, silencing of H19 represses these BCSC properties. In concordance, knockdown of H19 markedly inhibits tumor growth and suppresses tumorigenesis in nude mice. Mechanistically, we found that H19 functions as a competing endogenous RNA to sponge miRNA let-7, leading to an increase in expression of a let-7 target, the core pluripotency factor LIN28, which is enriched in BCSC populations and breast patient samples. Intriguingly, this gain of LIN28 expression can also feedback to reverse the H19 loss-mediated suppression of BCSC properties. Our data also reveal that LIN28 blocks mature let-7 production and, thereby, de-represses H19 expression in breast cancer cells. Appropriately, H19 and LIN28 expression exhibits strong correlations in primary breast carcinomas. Collectively, these findings reveal that lncRNA H19, miRNA let-7 and transcriptional factor LIN28 form a double-negative feedback loop, which has a critical role in the maintenance of BCSCs. Consequently, disrupting this pathway provides a novel therapeutic strategy for breast cancer.

Rationale and objectives Non-muscle-invasive bladder cancer (NMIBC) is highly recurrent, with each recurrence potentially progressing to muscle-invasive cancer, affecting patient prognosis. Intratumoral heterogeneity plays a crucial role in NMIBC recurrence. This study investigated a novel habitat-based radiomic analysis for stratifying NMIBC recurrence risk. Materials and methods A retrospective collection of 382 NMIBC patients between 2015 and 2021 from two medical institutions was carried out. Patients’ CT images were collected across three phases, with tumor sites delineated within the bladder. Intratumoral habitats were identified using K-means clustering on 19 texture features of the tumor sites, followed by the extraction of 107 radiomic features per habitat with PyRadiomics. These features were integrated into machine learning algorithms to develop a habitat-based model (HBM) for predicting two-year recurrence of NMIBC patients. The clinical and multiphase radiomic models were also constructed for comparison, with the Delong test comparing their diagnostic efficiency. The impact of HMB on patients’ recurrence-free survival and the correlation between HBM and tumor-stroma ratio were further analyzed. Results Three distinct habitats were identified within NMIBC. The HBM showed an AUC of 0.932 (95% CI: 0.906 - 0.958) in the training cohort and 0.782 (95% CI: 0.674 - 0.890) in the validation cohort for predicting two-year recurrence. With comparison between different models, The HBM is demonstrated to possess superior diagnostic efficacy to the clinical model (p < 0.001) in the training cohort. However, no significant difference was noted between the multiphase and clinical models (p = 0.130) in the training cohort. The HBM score effectively distinguished the recurrence-free survival of NIMBC patients and demonstrated a significant correlation with the tumor-stroma ratio. Conclusions Habitat-based radiomics, coupled with machine learning, efficiently predicts NMIBC recurrence. Further research on habitat-based radiomics offers potential improvement in clinical management of NMIBC.

Objective: Treating pediatric osteosarcoma in long bones is challenging due to skeletal immaturity, which restricts the generalizability of insights derived from adult patients. Are there disparities in outcomes? How should surgical protocols be tailored for children of varying ages? What are the specific postoperative complications? A large single-center retrospective cohort study of 345 patients under 14 years old with lower-limb osteosarcoma treated in our department since 2000 was conducted to address these inquiries. Methods: A retrospective analysis of 345 pediatric patients with lower-limb osteosarcoma admitted to our department between 2000 and 2019 was conducted. Clinical and functional outcomes were compared based on age groups, surgical methods, type of prosthesis, and primary tumor location. Patients were divided into the low-age group (<= 10 y old) and the high-age group (>10 y old). Overall survival rate (OS), progressionfree survival rate (PFS), and prosthesis survival rate were assessed using Kaplan-Meier curves, nonparametric survival analysis (log-rank test), and Univariate cox regression were used for comparison. The incidence of complications, local relapse rate (LRR), metastasis rate, final limb-salvage, and amputation rate, and Musculoskeletal Tumor Society (MSTS) score of different independent groups were further evaluated using chi 2 test or Fisher's exact test, and t-test was employed to evaluate the measurement data. Results: The average age of the patients was 11.10 +/- 2.32 years ranging from 4 to 14 y, with an average follow-up duration of 48.17 months. The 5, 10, and 15-year OS rates were 50.3%, 43.8%, and 37.9%, respectively. The progression-free survival rate was 44.8% at 5 years and 41.1% at 10 years. The final limb salvage rate was 61.45%, while the final amputation rate was 38.55%. The low-age group had a higher amputation rate compared with the high-age group (48.00% vs. 33.18%, P=0.009). The overall LRR was 9.28%, and the incidence of metastasis was 28.99%. The LRR of the limb-salvage group was higher than the amputation group (P=0.004). The low-age group experienced more prosthesis-related complications than the high-age group (P=0.001). The most common prosthesis-related complication in the low-age group was soft-tissue failure, while the periprosthetic infection was most frequent in the high-age group. The high-age group had a higher cumulative prosthesis survival compared with the low-age group (P=0.0097). Modular prosthesis showed better MSTS scores and higher cumulative prosthetic survival than expandable prosthesis in pediatric patients (P<0.05). Conclusion: Limb preservation in pediatric patients becomes increasingly efficacious with advancing age, while consideration of amputation is warranted for younger patients. The prevailing postoperative complications associated with prosthesis encompass soft tissue failure and periprosthetic infection. Younger patients diagnosed with lower limb osteosarcoma exhibit a heightened amputation rate and a greater incidence of prosthesis-related complications.