Rectal cancer (RC) is a malignant tumor originating in the rectum and represents a significant global health burden. As the eighth most common malignancy worldwide, RC accounts for approximately 340,000 deaths annually (1). The etiology of RC remains unclear but is thought to involve environmental factors, dietary habits, and genetic predispositions (2). Despite advancements in chemoradiotherapy and immunotherapy, the prognosis for RC patients has not significantly improved (3). Extensive research has identified various biomarkers associated with RC survival and prognosis. Notably, the advent of high-throughput sequencing has deepened our understanding of genetic alterations in RC, enabling the development of multigene predictive models using clinical and genetic data from public databases (4–8). Consequently, constructing an effective prognostic model on the basis of gene signatures is crucial for advancing personalized therapy and improving patient outcomes.

Temperature-sensitive receptor (TRP) channels are nonselective cation channels that respond to temperature changes and play key roles in thermoregulation, inflammation, pain modulation, and osmoregulation. Dysregulation of TRP channels has been linked to various diseases (9, 10). For instance, Zhao et al. classified esophageal squamous cell carcinoma patients into high- and low-risk groups on the basis of TRP-related prognostic gene scores, revealing elevated immune checkpoint expression in high-risk patients (11). These findings suggest that such patients may benefit more from immunotherapy. However, the prognostic significance of TRP channels in RC remains unexplored. Investigating their role could improve patient prognosis and increase survival rates.

In this study, we analyzed transcriptome data and clinical information from public databases to investigate RC. Using bioinformatics approaches, we identified five TRP-related genes and developed a predictive model, which was validated with external data from the Gene Expression Omnibus (GEO) database. Additionally, we explored the mechanistic differences between risk groups and examined the relevance of these key genes within the tumor immune microenvironment. Our findings provide valuable insights into the molecular mechanisms of RC and lay a theoretical foundation for future research.

Rectal cancer (RC) is frequently diagnosed at advanced stages because of the lack of distinctive early symptoms, which contributes to delayed detection and unfavorable clinical outcomes (19). This diagnostic challenge underscores the critical need for the discovery of novel biomarkers to enable risk stratification and personalized therapeutic interventions. While temperature-sensitive receptor (TRP) channels have been extensively implicated in diverse pathological processes, including tumorigenesis and immune modulation (9, 10), their prognostic relevance in RC remains largely uncharted. To bridge this knowledge gap, we systematically integrated multiomics data and bioinformatics approaches, revealing five TRP-related biomarkers (BMP5, DHRS11, GLTP, NFE2L3, and TMCC3) with potential diagnostic and therapeutic implications for RC. Importantly, these biomarkers not only align with known oncogenic pathways but also exhibit novel associations with RC-specific immune microenvironment alterations, suggesting new perspectives for the diagnosis and treatment of RC.

The five identified biomarkers play multifaceted roles in tumor biology, with both conserved and context-dependent functions across malignancies. BMP5, a member of the TGF-β superfamily, has tumor-suppressive effects on colorectal carcinogenesis. Our findings corroborate prior reports that BMP5 deficiency occurs in 7.7% of sporadic CRC cases and selectively predicts prognosis in this subtype (20).Mechanistically, the BMP5-mediated suppression of EPSTI1 via Jak-Stat signaling may represent CRC-specific vulnerability, potentially explaining its limited prognostic relevance in other tumors. DHRS11 has emerged as a pleiotropic regulator, with our data reinforcing its dual role in hormone-dependent cancers: while it sustains androgen receptor signaling in prostate cancer (21), its prognostic association in breast cancer (22) suggests tissue-specific modulation of steroid hormone pathways—Given that research has shown that female steroid hormones can reduce the risk of colorectal cancer (CRC) (23), we speculate that DHRS11 may affect CRC by regulating the steroid hormone pathway. However, there is currently a lack of sufficient evidence regarding the direct effect of DHRS11 on CRC, and this study is also the first attempt to link DHRS11 with CRC. In this case, our hypothesis still needs further in-depth research to support it. GLTP downregulation in CRC aligns with its tumor-suppressive function as a MIR196B target (24), indicating that lipid metabolism perturbations mediated by GLTP loss may drive RC progression. There are also studies indicating that overexpression of LTP can inhibit the growth of human colon cancer cells (HT-29; HCT-116), mainly by interfering with cell cycle progression and inducing cell necrosis (25). Our research findings indicate that GLTP expression is downregulated in the CRC group, further indirectly supporting its potential protective effect on CRC patients. Notably, NFE2L3 has paradoxical roles depending on the cellular context: although murine models suggest its anti-inflammatory and tumor-restraining effects in colon cancer (26), human studies indicate its pro-oncogenic functions in other malignancies. This dichotomy underscores the importance of microenvironmental influences on NFE2L3 activity. Meanwhile, studies have shown that NFE2L3 can regulate inflammation and oxidative stress-related genes in the colon, thereby keeping the microenvironment in a pro-inflammatory state (27). The occurrence of inflammation and oxidative stress often promote the occurrence and development of CRC (28). Our study also confirmed the upregulation trend of NFE2L3 in CRC patients, suggesting that inhibiting oxidative stress and inflammatory response may help alleviate the impact of NFE2L3 on CRC.TMCC3, previously characterized as a breast cancer stemness maintainer through AKT activation (29), has emerged as a potential RC progression driver, with our survival analysis linking its expression to adverse outcomes. The conserved tumor-promoting role of TMCC3 across malignancies suggests broad therapeutic targeting potential.

Through qRT-PCR experiments, we found that DHRS11, GLTP, and NFE2L3 have dysregulated expression in rectal cancer tissues. Survival analysis further established the status of BMP5, DHRS11, NFE2L3, and TMCC3 as independent prognostic indicators. It is worth noting that we found differences in the expression patterns of BMP5 and TMCC3 in RNA-seq and qRT-PCR validation, which may be attributed to the small sample size and heterogeneity between samples. However, it cannot be ignored that the results of qRT-PCR experiments and bioinformatics analysis jointly locate TRP-related biomarkers at the intersection of oncogenic signaling and metabolic reprogramming. In the future, we plan to expand the sample size and conduct larger-scale, multicenter studies, including clinical samples from different races and regions, to comprehensively study the changes of these biomarkers in vivo and provide more systematic insights for their application in clinical monitoring.

To increase the clinical utility of our prognostic model, we developed a nomogram that integrates risk scores with age, which significantly improved the accuracy of risk stratification. Comparative transcriptomic analysis between the high- and low-risk groups revealed differentially expressed genes (DEGs) predominantly enriched in cytokine–cytokine receptor interactions—pathway hubs governing tumor–stroma crosstalk. This finding aligns with emerging evidence that cytokine networks reprogram the tumor microenvironment to foster therapeutic resistance and metastatic dissemination.

Notably, CCL3 exemplifies the dual role of cytokines in CRC progression: while it recruits immune cells via chemotaxis, its overexpression activates the TRAF6/NF-κB axis to promote tumor cell survival and invasion (30). Similarly, IL-6/IL-11 signaling in cancer-associated fibroblasts (CAFs) induces STAT3 activation, creating a protumorigenic niche that drives CRC growth and correlates with dismal outcomes (31). Among chemokines, CXCL8 stands out as a master regulator of autocrine signaling in CRC. Its upregulation not only enhances tumor cell proliferation and anoikis resistance (32, 33) but also facilitates VEGF-independent angiogenesis and confers chemoresistance through mechanisms involving PI3K/Akt and MAPK pathway activation (34, 35). These observations collectively suggest that high-risk RC patients may exhibit hyperactivated cytokine signaling, rendering them susceptible to microenvironment-driven progression.

Importantly, our model’s incorporation of cytokine-related DEGs provides a mechanistic link between TRP-associated genetic signatures and immune-metabolic dysregulation, offering actionable targets for intercepting cytokine-mediated malignant transformation.

Our immune profiling uncovered distinct immune landscapes between risk groups, characterized by significant differences in five immune cell subsets. CD8⁺ T cells, pivotal effectors of antitumor immunity, mediate tumor cell lysis through granzyme/perforin release and interferon-γ secretion at immune synapses (36, 37). Conversely, type 2 T helper (Th2) cells exhibit protumoral properties: IL-4/STAT6/GATA3 signaling drives Th2 polarization and subsequent secretion of IL-5/IL-13, which collectively promote metastatic spread (38, 39). It is worth noting that in this study, TMCC3 was found to be strongly positively correlated with central memory CD8+T cells, while DHRS11 was strongly negatively correlated with type2 T helper cells. The identified biomarkers may significantly influence the immune response of RC, thereby implying their potential significance in the prevention and treatment of RC at the cytokine level. In addition, immune checkpoint molecules PD-1 (PDCD1), PD-L1 (CD274), and CTLA4 mediate tumor immune escape by suppressing the activity of immune cells and are known potential targets for RC immunotherapy (40). Studies have shown that activated CD8⁺ T cells highly express PD-1 under continuous antigen stimulation (41). Based on these research findings, we speculate that the high expression of TMCC3 may indirectly induce the upregulation of PD-1 by stimulating the high expression of CD8⁺ T cells, thereby promoting tumor progression. Therefore, patients with this characteristic may be more sensitive to PD-1 inhibitors due to the enrichment of PD-1⁺ CD8⁺ T cells. The CTLA-4 axis affects tumors by altering the Th1/Th2 balance (42). Based on this, we believe that the negative correlation between DHRS11 and Th2 cells may act on the CTLA4 axis by influencing the Th1/Th2 balance, and its low expression may relieve the inhibition on Th2 cells, promoting the increased secretion of Th2-type cytokines, forming an immune escape microenvironment, and thereby affecting tumors. These mechanisms still need further verification, but they initially reveal that TRP-related genes may act through a regulatory network formed by immune cell infiltration and checkpoint molecules. They also suggest that TMCC3 and DHRS11 may serve as potential markers for predicting the response to immunotherapy, providing a new basis for precise stratification in RC immunotherapy.

To elucidate the intrinsic relationship between the five biomarkers and the classical RAS/RAF axis in colorectal cancer (RC), a correlation analysis was conducted. This analysis identified the strongest positive correlation between TMCC3 and KRAS, while GLTP and HRAS exhibited a negative correlation. KRAS, recognized as one of the most frequently mutated genes in colorectal cancer, significantly influences patient prognosis and survival, and serves as a potential therapeutic target (43). Concurrently, TMCC3 is known to sustain cancer stem cell properties by activating the AKT pathway (29). Considering the frequent crosstalk between the PI3K/AKT and RAS/RAF/ERK pathways, and given that Akt/Ras/Raf/MEK/ERK are established therapeutic targets in cancer (44), it is plausible that the coordination between KRAS and TMCC3, potentially through AKT activation, may enhance the RAS/RAF axis of cancer signaling, thereby promoting tumor progression. Furthermore, as HRAS is a common mutation site in RC (45), the observed negative correlation between HRAS and GLTP may be associated with the tumor suppressor mechanism of GLTP. Overexpression of GLTP can induce cell cycle arrest at the G1/S checkpoint by upregulating p27 and p21 (26), thereby inhibiting Ras and blocking the cell cycle.

Targeted drugs predictions were performed using the Genomics of GDSC database, which identified 138 targeted drugs associated with RC. Among these, 36 drugs have significant differences between high- and low- risk patient groups. In addition, the top 5 drugs, including MK.2206, Pazopanib, JNK.lnhibitor.VIII, PLX4720 and NU.7441 were shown to be more sensitive in high-risk group. The utilization of these drugs helps improve the prognosis of patients in this group (46–52). In conclusion, these findings provide valuable insights into potential therapeutic targets for RC treatment.

Our study establishes TRP-related biomarkers as multidimensional regulators of RC progression, orchestrating oncogenic signaling, immune evasion, and therapeutic vulnerability. While further preclinical validation is needed, these findings illuminate a precision oncology framework where TRP-based stratification guides the selection of targeted agents (e.g., AKT inhibitors) and immunomodulators, ultimately bridging genomic insights with clinical actionability.

Although we have adopted a series of advanced bioinformatics analysis methods, mainly focusing on the mining of transcriptome data, single data mining may lead to incomplete understanding of the results. In the future, we plan to combine proteomics, metabolomics and other omics data to further explore potential biomarkers and therapeutic targets of TRP related genes, providing a more comprehensive basis for accurate diagnosis and treatment of rectal cancer. In addition, although we have experimentally validated the expression of some biomarkers through qRT PCR, these experiments have only been conducted in limited samples and have not yet undergone further functional validation. Subsequent research will further explore the specific molecular mechanisms of these five TRP related genes in rectal cancer, such as studying their effects on the proliferation, apoptosis, invasion, and metastasis of rectal cancer cells through in vitro cell experiments and in vivo animal models, as well as their interactions with immune cells, cytokines, and other factors in the tumor microenvironment, in order to gain a more comprehensive understanding of their roles in the occurrence and development of rectal cancer. Finally, we have not yet delved into the clinical application potential of these biomarkers, which limits their widespread use in clinical settings. In the future, we will conduct relevant clinical trials to explore the clinical application value of these biomarkers and provide personalized treatment plans for clinical practice. In summary, in the future, we will further reveal the role of TRP related genes in rectal cancer through multi-level research and promote their practical application in clinical practice.

This study identified a TRP channel-related gene signature (BMP5, DHRS11, GLTP, NFE2L3, and TMCC3) that predicts rectal cancer prognosis and modulates tumor–immune crosstalk. The integrative risk model, validated through multiomics analyses, enables patient stratification for targeted therapies (e.g., AKT/BRAF inhibitors) and immunomodulation strategies. These findings bridge TRP biology with precision oncology, offering a roadmap for biomarker-driven RC therapeutics.