Objective To explore the prognostic value of glycolysis-related genes in colorectal cancer (CRC) patients and formulate a novel glycolysis-related prognostic risk model. Methods Single-cell and bulk transcriptomic data of CRC patients, along with clinical information, were obtained from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Glycolysis scores for each sample were calculated using single-sample Gene Set Enrichment Analysis (ssGSEA). Kaplan-Meier survival curves were generated to analyze the relationship between glycolysis scores and overall survival. Novel glycolysis-related subgroups were defined among the cell type with the highest glycolysis scores. Gene enrichment analysis, metabolic activity assessment, and univariate Cox regression were performed to explore the biological functions and prognostic impact of these subgroups. A prognostic risk model was built and validated based on genes significantly affecting the prognosis. Gene Set Enrichment Analysis (GSEA) was conducted to explore differences in biological processes between high- and low-risk groups. Differences in immune microenvironment and drug sensitivity between these groups were assessed using R packages. Potential targeted agents for prognostic risk genes were predicted using the Enrichr database. Results Tumor tissues showed significantly higher glycolysis scores than normal tissues, which was associated with a poor prognosis in CRC patients. The highest glycolysis score was observed in epithelial cells, within which we defined eight novel glycolysis-related cell subpopulations. Specifically, the P4HA1⁺ epithelial cell subpopulation was associated with a poor prognosis. Based on signature genes of this subpopulation, a six-gene prognostic risk model was formulated. GSEA revealed significant biological differences between high- and low-risk groups. Immune microenvironment analysis demonstrated that the high-risk group had increased infiltration of macrophages and tumor-associated fibroblasts, along with evident immune exclusion and suppression, while the low-risk group exhibited higher levels of B cell and T cell infiltration. Drug sensitivity analysis indicated that high-risk patients were more sensitive to Abiraterone, while low-risk patients responded to Cisplatin. Additionally, Valproic acid was predicted as a potential targeted agent. Conclusion High glycolytic activity is associated with a poor prognosis in CRC patients. The novel glycolysis-related prognostic risk model formulated in this study offers significant potential for enhancing the diagnosis and treatment of CRC.
Humans ; Colorectal Neoplasms/pathology* ; Glycolysis/genetics* ; Prognosis ; Transcriptome ; Tumor Microenvironment/genetics* ; Gene Expression Profiling ; Single-Cell Analysis ; Gene Expression Regulation, Neoplastic ; Male ; Female ; Kaplan-Meier Estimate

Objective To explore the effects of estrogen on stress-induced senescence of vascular smooth muscle cells (VSMCs) and the underlying mechanisms. Methods The VSMCs of passage 2-3 cultured from female SD rats were induced into senescence by exposing to 150μmol/L H2O2 in the presence or absence of different concentrations(10-10mol/L-10-8mol/L) of 17β-estradiol (E2). The expressions or activities of senescence associated marker DcR2, senescence-associated beta-galactosidase (SA-β-Gal), oncogene Ras and p21WAF1 were detected by flow cytometry, cytochemical staining, pull-down assay or Western blotting analysis. Results Flow cytometry analysis showed that in the physiological concentrations, E2 significantly inhibited the H2O2-promoted high-level expression of DcR2 of VSMCs in a dose-dependent manner, with a highest inhibitive rate at 14.48%±0.6%(E2=10-8 mol/L;P<0.05, n =3);this inhibitive effect could be blocked by a E2 receptors inhibitor ICI 182,780. Cytochemistry staining showed that the rate of SA-β-Gal positive VSMCs induced by H2O2 decreased in presence of 10-8mol/L E2 (20.5%±1.4% vs 9.6%±0.9%;P<0.05, n =9). Pull-down assay and Western blotting analysis revealed that administration of 10-8mol/L E2 obviously reduced the H2O2-induced activity of Ras (0.60±0.06 vs 0.26±0.04;P<0.05, n =3) and expression of p21WAF1 (0.46±0.04 vs 0.33±0.02;P<0.05, n =3). Conclusion E2 exerts, an inhibitive effects on stress-induced senescence of VSMCs by suppressing the activity of Ras and expression of p21WAF1. This finding suggests a novel mechanism for the hormone's anti-atheroschlerotic effects.