Objective: To investigate whether aurora kinase A (AURKA) in hepatocellular carcinoma (HCC) cells drives M2 polarization of macrophages via exosomes, and to clarify its underlying molecular mechanism.
Methods: The correlations between AURKA expression level in HCC tissues and M2 macrophage infiltration as well as prognosis of HCC patients were analyzed using the GEPIA (Gene Expression Profiling Interactive Analysis) and TIMER (Tumor Immune Estimation Resource) public databases. Based on the human HCC cell line MHCC-97H, AURKA stably overexpressed (97H-AKAOE) and knockdown (97H-AKAKD) cells were constructed. Exosomes were isolated by ultracentrifugation and characterized by transmission electron microscopy, nanoparticle tracking analysis, and Western blotting. Exosomes derived from 97H-AKAOE and 97H-AKAKD cells were co-cultured with M0 macrophages, and the expression levels of M2 macrophage marker proteins CD163 and Arg-1 were assessed by real-time quantitative reverse transcription PCR (RT-qPCR) and Western blotting to evaluate the effect of AURKA on M2 polarization of macrophages. Liquid chromatography-mass spectrometry (LC-MS) was used to identify key differentially expressed proteins regulated by AURKA in exosomes, and functional rescue experiments were performed to validate their roles on M2 polarization of macrophages.
Results: AURKA was highly expressed in HCC tissues, and its expression level positively correlated with poor prognosis of HCC patients and M2 macrophage infiltration abundance (all P < 0.05). AURKA overexpression significantly enhanced the exosome-induced M2 polarization of macrophages, as evidenced by the upregulated CD163 and Arg-1, whereas AURKA knockdown suppressed this effect. Proteomic analysis revealed that lectin galactoside-binding soluble 3-binding protein (LGALS3BP) was significantly enriched in exosomes from AURKA-overexpressing HCC cells. Knockdown of LGALS3BP partially reversed the AURKA-mediated M2 polarization of macrophages.
Conclusion: AURKA in HCC cells drives M2 polarization of macrophages by upregulating exosomal LGALS3BP expression, thereby promoting the formation of an immunosuppressive tumor microenvironment in HCC. Elucidation of this mechanism provides a potential novel target for HCC immunotherapy.
Objective: To investigate the correlations between changes in cervical microcirculation parameters and ovarian reserve function after transumbilical single-port laparoscopic myomectomy.
Methods: A total of 165 patients diagnosed with uterine fibroids at the 73rd Group Army Hospital of the People’s Liberation Army from February 2022 to January 2025 were enrolled. Based on surgical approaches, patients were divided into three groups: the open abdominal group (55 cases, open abdominal myomectomy), the multiport group (55 cases, traditional multiport laparoscopic myomectomy), and the single-port group (55 cases, transumbilical single-port laparoscopic myomectomy). General patient information and perioperative indicators were collected. Pain intensity was assessed using the Visual Analog Scale (VAS) immediately after surgery and at 12, 24, and 48 h postoperatively. At discharge, patient satisfaction with the incision was evaluated using the Cosmetic Scale (CS) and Body Image Scale (BIS). Postoperative recovery status and complications were recorded. Cervical microcirculation parameters, including microvessel diameter, blood flow perfusion, and capillary diameter, were measured preoperatively and at 12, 24, and 72 h postoperatively. One-way analysis of variance (ANOVA) was used to compare differences in cervical microcirculation parameters before and after surgery, while multivariate ANOVA was used to compare differences among the three surgical approaches at 72 h postoperatively. Ovarian reserve function indices, including serum follicle-stimulating hormone (FSH), anti-Müllerian hormone (AMH), luteinizing hormone (LH), peak systolic velocity (PSV) of ovarian stromal artery, estradiol (E2), antral follicle count (AFC), and ovarian volume, were assessed preoperatively and 3 months postoperatively. A difference-in-differences (DID) univariate model was constructed to analyze the differences in ovarian reserve function indices before and after surgery, with independent t-tests comparing differences among the three surgical approaches at 3 months postoperatively. Repeated-measures ANOVA and DID full model analysis were employed to evaluate the effects of surgical approach, postoperative time, and their interaction (group×time) on cervical microcirculation parameters and ovarian reserve function-related indices. Pearson correlation analysis was performed to evaluate the correlations between changes in cervical microcirculation parameters and changes in ovarian reserve function indices after surgery.
Results: Regarding postoperative pain, VAS scores in all three groups showed a gradual declining trend over time, with the single-port group demonstrating the lowest scores at all time points, followed by the multiport group. Concerning postoperative incision satisfaction, compared with the open abdominal group, patients in the multiport and single-port groups showed significantly lower BIS scores and significantly higher CS scores (all P < 0.05). Compared with the multiport group, patients in the single-port group demonstrated even lower BIS scores and higher CS scores (all P < 0.05). Regarding postoperative recovery and complications, the rates of fibroid recurrence and menstrual cycle disturbance within 1 year postoperatively in the multiport and single-port groups were significantly lower than those in the open abdominal group (all P < 0.05); additionally, the total complication rate in the single-port group was significantly lower than that in the open abdominal group (P < 0.05). There were no statistically significant differences between the multiport and single-port groups regarding the aforementioned indices and postoperative pregnancy rate (all P < 0.05). Moreover, the total complication rates between the open abdominal and multiport groups showed no significant difference (P > 0.05). Repeated-measures ANOVA results showed that both surgical approaches and postoperative time had significant effects on cervical microcirculation parameters in all three groups (all P < 0.001), with the single-port group demonstrating significantly higher cervical microcirculation parameters at all postoperative time points compared to the multiport and open abdominal groups (all P < 0.05). Postoperatively, In all three groups, LH and FSH levels increased significantly, while E2, AMH, ovarian volume, AFC, and PSV decreased significantly. Notably, the magnitude of change in these indices in the open abdominal group was greater than those in the multiport and single-port groups. DID full model analysis indicated that both surgical approach and postoperative time had significant effects on ovarian reserve function in all three groups (all P < 0.001); however, this effect showed no significant difference between the multiport and single-port groups. Correlation analysis showed that the single-port group exhibited the strongest correlation between changes in cervical microcirculation parameters and changes in ovarian reserve function indices compared with the open abdominal and multiport groups (P < 0.008 after Bonferroni correction). Specifically, changes in blood flow perfusion (Δblood flow perfusion) showed relatively high correlation coefficients with changes in E2 (ΔE2), AMH (ΔAMH), and AFC (ΔAFC), with values of 0.56, 0.53, and 0.50, respectively.
Conclusion: Transumbilical single-port laparoscopic myomectomy is an effective treatment for uterine fibroids. Compared with traditional surgical approaches, single-port laparoscopy demonstrates more pronounced minimally invasive disadvantages and can effectively reduce the impact on ovarian reserve function by maintaining cervical microcirculation homeostasis, thus demonstrating good clinical application value.
Objective: To investigate the correlation between the dynamic changes in serum carbohydrate antigen 125 (CA125) levels and immune profiles as well as clinical prognosis in patients with recurrent ovarian cancer receiving bevacizumab combined with paclitaxel and carboplatin chemotherapy, with the aim of providing a basis for individualized treatment and prognostic evaluation.
Methods: A total of 90 patients with recurrent ovarian cancer admitted to the Cancer Hospital of Shantou University Medical College between June 2019 to June 2023 were enrolled in this study. Serum levels CA125, immunoglobulin A (IgA), IgG, IgM, complement component 3 (C3), and C4 were measured at the following time points: before chemotherapy (T0), at week 3 (T1), week 6 (T2), and week 9 (T3) of chemotherapy. A latent class growth model (LCGM) combined with a growth mixture model (GMM) was applied to characterize the trajectory of CA125 level changes throughout the chemotherapy course. Differences in demographic characteristics and immunological indicators across trajectory groups were compared, and generalized estimating equations (GEE) were used to examine the longitudinal associations between CA125 trajectory patterns and immunological indicators. Multivariate logistic regression was then employed to identify factors influencing CA125 level changes during chemotherapy, and COX regression analysis was conducted to assess the association between CA125 trajectory patterns and unfavorable prognosis.
Results: In 90 patients with recurrent ovarian cancer receiving bevacizumab combined with taxane-based and carboplatin chemotherapy, serum CA125 levels showed a progressive downward trend as chemotherapy advanced. The mean serum CA125 levels at T0, T1, T2, and T3 were 689.35±121.34 U/mL, 516.67±168.37 U/mL, 140.13±39.42 U/mL, and 30.12±9.27 U/mL, respectively (P < 0.05). Based on the trajectory of CA125 changes, patients were classified into three groups: the stable group (n=32, 35.56%), characterized by a gradual change in CA125 levels from T0 to T3 (β=−0.15, P<0.001); the rapid decline group (n=37, 41.11%), characterized by a sharp decrease in CA125 levels from T0 to T3 (β=−1.54, P<0.001); and the slow decline group (n=21, 23.33%), characterized by a steady decrease in CA125 levels from T0 to T3 (β=−0.82, P<0.001). Age≥50 years, serous histological type, and unilateral ovarian involvement were identified as independent factors associated with a rapid decline in CA125 levels during chemotherapy (all P<0.05), while serous histological type and unilateral ovarian involvement were also independently associated with a slow decline in CA125 levels (both P<0.05). At each time point from T1 to T3, the levels of IgA, IgG, IgM, C3, and C4 in the stable group were significantly higher than those in both the rapid decline group and the slow decline groups (all P<0.05). GEE analysis revealed that, compared with the stable group, the CA125 levels in both the rapid decline group and the slow decline group were negatively correlated with all the aforementioned immunological indicators (all P<0.05). COX regression analysis showed that patients in the rapid decline group had a higher risk of unfavorable prognosis than those in the stable group (HR=1.624), whereas patients in the slow decline group had a lower risk of unfavorable prognosis (HR=0.447); these associations remained consistent across all subgroups (P<0.05).
Conclusion: During bevacizumab combined with paclitaxel and carboplatin chemotherapy, CA125 level changes in patients with recurrent ovarian cancer exhibit significant heterogeneity, with notable differences in immune profiles and prognostic outcomes across different CA125 trajectory groups. Compared with patients in the stable group, those in the rapid decline group demonstrate relatively weaker immune function and a higher risk of unfavorable prognosis, whereas patients in the slow decline group exhibited a relatively better immune status and a lower risk of unfavorable prognosis. Dynamic monitoring of CA125 trajectory patterns throughout chemotherapy may provide clinically valuable reference for assessing immune status and predicting prognosis in patients with recurrent ovarian cancer.
Objective: This study aimed to explore the synergistic inhibitory effect of apatinib combined with PD-1 monoclonal antibody (mAb) on subcutaneous tumor growth in microsatellite-stable (MSS) colon cancer mice.
Methods: Using TCGA database, we analyzed clinical data and RNA-seq data from colon cancer patients, calculated angiogenesis scores via ssGSEA algorithm, and evaluated their correlation with clinicopathological features and prognosis. ssGSEA and CIBERSORT algorithms were employed to assess the relationship between immune cell infiltration and angiogenesis score. Subcutaneous tumors were established in mice using the CT26 cell line, with comparisons made between combination therapy (apatinib＋PD-1mAb) and monotherapy groups. Flow cytometry, immunohistochemistry, and multiplex immunofluorescence staining were used to analyze immune cell infiltration and angiogenesis differences.
Results: High angiogenesis score correlated with poor prognosis (P = 0.019), advanced N stage, and increased infiltration of M2 macrophages (R = 0.28, P < 0.01). In animal experiments, the combination group exhibited significantly inhibited tumor growth compared to controls and monotherapy groups (P < 0.05 for all comparisons). The tumor volume (Mean±SD) in the combined therapy group was 181.14±118.59 mm3, compared to 1000.71±254.69 mm3 in the control group, 676.14±392.52 mm3 in the apatinib monotherapy group, and 579.79±259.41 mm3 in the PD-1 monoclonal antibody group. Immunological analyses revealed significantly increased CD8+ T cell infiltration and reduced M2 macrophage infiltration in the combination group (P < 0.05).
Conclusion: These findings suggest that apatinib combined with PD-1 mAb may be a promising therapeutic strategy for “immune-desert” MSS colon cancer by significantly reducing tumor burden.Objective: Low protein diet can inhibit the occurrence and development of tumor, but its mechanism remains to be further explored. Based on the genes regulated by low protein diet, this project constructed and verified a risk model of low protein diet affecting the prognosis of colorectal cancer.
Methods: The differential genes between low protein diet and normal protein diet mice were screened in GEO dataset, and the risk model was constructed by univariate COX regression, LASSO regression analysis and multivariate COX regression analysis. The risk model of low protein diet was verified by TCGA and GEO databases. Using single sample gene set enrichment analysis to evaluate the enrichment degree of immune function related genes in high and low-risk groups. The “CIBERSORT” algorithm was used to analyze the enrichment of 22 immune cells in the tumor microenvironment. Using genomics prediction models to calculate IC50 values of common drugs in high and low-risk groups and evaluate targeted drug sensitivity.
Results: Six genes were screened to predict the prognosis of colorectal cancer. The risk score＝Exp（ASAH1）×（−2.089）＋Exp（CAMK2N1）×（−1.576）＋Exp（SPINK4）×（−0.232）＋Exp（REG3G）×0.607＋Exp（CTSD）×1.898＋Exp（STK25）×1.980. According to this model, mice fed a low protein diet had lower risk scores. The proportion of memory CD4+ T cells and dendritic cells was higher in low-risk group, and the proportion of macrophages was higher in high-risk group. Sensitive drugs for low-risk group: WZ4003, vorinostat, PRIMA-1MET, PLX-4720, pictilisib; sensitive drugs for high-risk group: BI-2536, KU-55933, NU744, RO-3306.
Conclusion: The risk model based on low protein diet can effectively predict the risk and prognosis of colorectal cancer, which can provide theoretical reference for the clinical diagnosis and treatment of colorectal cancer.Objective: Esophageal carcinoma (ESCA) ranks among the world's most aggressive and lethal malignancies. Histologically, it is classified into esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC), with ESCC accounting for 60%-70% of cases. Neoadjuvant radiotherapy is a crucial treatment modality for locally advanced ESCC. However, ESCC exhibits significant heterogeneity in radiosensitivity, and effective biomarkers for radiosensitivity remain elusive. This study aims to identify biomarkers for ESCC radiosensitivity and elucidate their functional roles through bioinformatics analysis.
Method: Given that ESCC is the predominant subtype of ESCA, we prioritized ESCC for biomarker analysis and functional investigation. Using the TCGA-ESCC dataset, patients were stratified into radiotherapy-sensitive and radiotherapy-resistant groups based on complete response (CR) status after radiotherapy. We investigated differentially expressed genes influencing radiotherapy sensitivity (CR) and resistance (non-CR) in esophageal cancer patients. Performed pathway enrichment analysis on differentially expressed genes using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases; Risk factors were screened using univariate and multivariate COX regression analyses to construct a prognostic risk prediction model for ESCC. Concurrently, pathway enrichment levels between high- and low-risk groups were assessed via Gene Set Enrichment Analysis (GSEA). Transcriptome expression data from ESCC were analyzed for immune infiltration using ESTIMATE, CIBERSORT, and ssGSEA algorithms. Protein interaction networks of IDO1 were analyzed using the STRING database (https://string-db.org/).
Results: Differentially expressed genes between radiotherapy-sensitive and resistant groups showed significant enrichment in immune-related pathways. Compared to the sensitive group, the resistant group exhibited a more active immune microenvironment, primarily manifested by a higher proportion of CD4+ T cells. IDO1 was highly expressed in the radiotherapy-resistant group, significantly correlated with poor prognosis, and possessed independent prognostic predictive ability. Additionally, the IDO1-high group exhibited heightened immune activation, suggesting IDO1 may regulate the immune microenvironment through tryptophan metabolism to contribute to esophageal cancer radiotherapy resistance.
Conclusion: This study confirms IDO1 as a potential key regulator involved in radiotherapy resistance and immune regulation in ESCC, holding significant biomarker [4] value and therapeutic target potential.Colorectal cancer (CRC) is a malignant tumor with high incidence and mortality rates worldwide, influenced by multiple factors including genetics, epigenetics, and tumor microenvironment. Traditional omics research, based on mixed cell samples, can only obtain average signals from cell populations, making it difficult to resolve the high heterogeneity within tumors. The emergence of single cell sequencing technology provides a powerful tool for revealing the cellular lineages, molecular characteristics, and tumor microenvironment of CRC at single cell resolution. This article systematically reviews the development history and core principles of single cell sequencing technology, with a focus on summarizing recent advances in CRC tumor heterogeneity, tumor microenvironment, mechanisms of drug resistance and metastasis, and clinical translational applications. Furthermore, it discusses the current challenges in technology and data analysis at the single cell level, and provides future development directions for single cell sequencing in multi-omics integration and precision oncology.
