CD137L and colorectal cancer prognosis: Insights from clinical and TCGA data analysis.

BackgroundmiRNA-154 (miR-154) has been identified as a tumor suppressor in several types of human cancers. However, its clinical significance in colorectal cancer (CRC) is still unclear. The aim of this study was to analyze the association of miR-154 expression with clinicopathologic features and prognosis in CRC patients.MethodsQuantitative RT-PCR was performed to evaluate miR-154 levels in 169 pairs of CRC specimens and adjacent noncancerous tissues. Then, the associations of miR-154 expression with clinicopathological factors or survival of patients suffering CRC were determined.ResultsThe expression levels of miR-154 in CRC tissues were significantly lower than those in corresponding noncancerous tissues (P < 0.001). Decreased miR-154 expression was significantly associated with large tumor size, positive lymph node metastasis, and advanced clinical stage. Moreover, the univariate analysis demonstrated that CRC patients with low miR-154 expression had poorer overall survival (P = 0.006). The multivariate analysis identified low miR-154 expression as an independent predictor of poor survival.ConclusionsThese findings suggested that miR-154 downregulation may be associated with tumor progression of CRC, and that this miR may be an independent prognostic marker for CRC patients.

Objective To study the expressions of Eag1 and vascular endothelial growth factor (VEGF) in colorectal cancer, and to explore the relationships between Eag1 and VEGF expressions and clinicopathologic features of colorectal cancer. The correlation between Eag1 and VEGF and their clinical significance in colorectal cancer were explored. Methods A total of 73 cases of colorectal cancer tissues, 25 cases of colorectal adenoma and 10 cases of normal colorectal mucosa from January 2015 to December 2016 in Hunan Provincial People′s Hospital were collected. The expressions of Eag1, VEGF protein and mRNA in 73 cases of colorectal cancer tissues, 25 cases of colorectal adenoma and 10 cases of normal colorectal mucosa were detected by immunohistochemistry and real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). The relationships between the two factors and the clinical pathological features of patients with colorectal cancer were analyzed. Results Immunohistochemistry showed that the positive rate of Eag1 in colorectal cancer group was 86.30% (63/73), which was significantly higher than that in colorectal adenoma group (20.00%, 5/25) and normal colorectal mucosa group (10.00%, 1/10), with significant differences (χ2=38.539, P<0.001; χ2=24.874, P<0.001). The positive rate of VEGF in colorectal cancer group was 89.04% (65/73), which was significantly higher than that in colorectal adenoma group (56.00%, 14/25) and normal colorectal mucosa group (30.00%, 3/10), with significant differences (χ2=10.980, P<0.001; χ2=16.911, P<0.001). There was a significant positive correlation between Eag1 and VEGF in colorectal cancer and colorectal adenoma tissues (c=0.580, P=0.010; c=0.669, P<0.001). The results of RT-qPCR showed that the expressions of Eag1 and VEGF mRNA in colorectal cancer group, colorectal adenoma group and normal colorectal mucosa group were 4.50±0.34, 3.14±0.44; 1.42±0.22, 1.61±0.15 and 1.00±0.16, 1.00±0.53, respectively, and the differences were statistically significant (F=15.546, P=0.014; F=5.363, P=0.025). The relative expression levels of Eag1 and VEGF mRNA in colorectal cancer group was about 4.50 times (t=31.851, P=0.003) and 3.14 times (t=6.870, P=0.014) than those in normal colorectal mucosa group. The mRNA expression levels of Eag1 and VEGF in colorectal cancer were correlated with degree of differentiation (F=840.725, P<0.001; F=102.950, P<0.001), depth of invasion (t=4.754, P<0.001; t=8.557, P<0.001), Duke staging (F=179.902, P<0.001; F=55.911, P<0.001) and lymph node metastasis (t=20.337, P<0.001; t=25.218, P<0.001). But they were not related to age (t=0.196, P=0.845; t=0.534, P=0.595) and sex (t=0.409, P=0.684; t=1.078, P=0.285). Conclusion Eag1 and VEGF are highly expressed in colorectal cancer tissues with positive correlation, and are closely related to the degree of differentiation, depth of invasion, Duke staging and lymph node metastasis of colorectal cancer. Both of them play important roles in the occurrence and development of colorectal cancer, which is expected to provide favorable research directions to the early diagnosis, treatment and prognosis of colorectal cancer. Key words: Colorectal neoplasms; Vascular endothelial growth factors; Eag1

To investigate the microRNA101(miR101) expression and its clinical significance in colorectal cancer. Tissue microarrays containing 56 specimens of normal mucosa, 51 adenoma and 735 colorectal cancer were examined by locked nucleic acid in-situ hybridization(LNA-ISH) for miR101 expression. Relationship between miR101 expression and clinicopathologic parameters and prognosis of colorectal cancer patients were analyzed. Fresh frozen tissues containing 5 specimens of normal mucosa, 5 adenoma and 47 colorectal cancer were examined by RT-PCR to verify the accuracy of LNA-ISH. Expression of miR101 increased gradually from normal mucosa, adenoma to stage I colorectal cancer (all P<0.01), and decreased gradually from stage II(, stage III( to stage IIII( colorectal cancer (all P<0.01). Overexpression of miR101 was related with lower incidence of lymph node metastasis, lower metastasis rate, higher differentiation and lower recurrence rate (all P<0.01). Multivariate survival analysis demonstrated that miR101 expression was an independent prognostic factor of overall survival (HR=0.550, 95% CI: 0.351-0.863) and disease free survival(HR=0.562, 95% CI: 0.397-0.794) in colorectal cancer patients. Overexpression of miR101 predicted a better prognosis in colorectal cancer patients. Expression of miR101 is associated with the genesis and development of colorectal cancer, and may serve as an independent prognostic factor for colorectal cancer patients.

Keratin-15 (KRT15) is a type I keratin lacking a defined type II partner and plays a key role in maintaining cytoplasmic stability. Recently, studies have reported that KRT15 was correlated with tumor formation and progression. However, the clinical significance of KRT15 in colorectal cancer is unclear. In this study, we aimed to investigate the expression of KRT15 and its clinical significance in colorectal cancer. KRT15 expression was examined in 98 cases of colorectal cancer and matched adjacent normal tissues by quantificational real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC), respectively. Then, the clinical significance of KRT15 expression was evaluated in colorectal cancer. QRT-PCR results revealed that the mRNA levels of KRT15 in colorectal cancer tissues were significantly higher compared with those in normal tissues (p<0.0001). The rates of KRT15 high-expression in colorectal cancer and normal tissues were 57.1% and 8.9%, respectively, and the difference was statistically significant (p<0.0001). KRT15 high-expression correlated with differentiation, T stage, lymph node metastasis and clinical stage in colorectal cancer (p<0.05). Meanwhile, KRT15 overexpression predicted poor prognosis and could be used as an independent prognostic factor. These data indicate KRT15 is highly expressed in colorectal cancer and may serve as a prognostic biomarker.

A Master microRNA miR-508-3p Modulates the Mesenchymal Subtype of Colorectal Cancer by Targeting ZEB1/BMI1/SALL4 Network

BackgroundThe clinical significance of LINC00996 in colorectal cancer (CRC) has not been verified. In the current study, the authors aimed to explore the expression of LINC00996 and its clinical significance in CRC based on the data mining of Gene Expression Omnibus (GEO) and the Cancer Genome Atlas (TCGA) datasets, as well as to elucidate the functions of its potential target genes.Materials and methodsGEO and TCGA microarray datasets were used to evaluate the LINC00996 expression and its clinical significance in CRC. LINC00996 related genes were identified by Multi Experiment Matrix, RNA-Binding Protein DataBase, and The Atlas of Noncoding RNAs in Cancer. Subsequently, they were sent to gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analysis.ResultsLINC00996 is significantly decreased in CRC tissues compared with non-tumor tissues. Low level of LINC00996 is associated with remote metastasis and poor overall survival. However, LINC00996 has a minimal effect on gender, lymphatic invasion, tumor size, lymph node metastasis, and pathological stage. One hundred and forty-two LINC00996 related genes were identified; the results of functional analysis indicated that LINC00996 might repress tumorigenesis and metastasis via modulating the JAK-STAT, NF-κB, HIF-1, TLR, and PI3K-AKT signaling pathways.ConclusionOur study demonstrates that decreased LINC00996 expression may be involved in colorectal carcinogenesis and metastasis, and the depletion of LINC00996 is associated with a poor outcome in CRC patients. Moreover, the JAK-STAT, NF-κB, HIF-1, TLR, and PI3K-AKT pathways may be the key pathways regulated by LINC00996 in CRC.

The aim of this study was to identify genes with clinical significance in colorectal cancer (CRC). Gene expression profiles of 585 CRC tissues and 61 normal colorectal tissues from Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases were used to identify differentially expressed genes (DEGs) between CRC and normal colorectal tissues. DAVID and KOBAS tools were used to explore Gene Ontology (GO) and KEGG pathways enriched by DEGs, respectively. In addition, TCGA data sets were also used to identify prognostic factors and develop a prognostic prediction model for CRC. A total of 353 DEGs including 117 upregulated and 236 downregulated genes in CRC were identified based on GSE32323 data set. These DEGs were significantly enriched in the biological process related to the regulation of cell proliferation and 50 signaling pathways, such as "TGF-beta signaling pathway," "Wnt signaling pathway," and "Jak-STAT signaling pathway." GCG, ADH1B, SLC4A4, ZG16, and CLCA4 were the top five downregulated in CRC. FOXQ1, LGR5, CLDN1, KRT23, and DPEP1 were the top five upregulated in CRC. KRT23 expression could affect tumor stage and regional lymph node metastasis in CRC patients. FOXQ1 expression could affect tumor distant metastasis in CRC patients. Survival analysis indicated that SLC4A4 expression was associated with the prognosis of CRC patients. Prognostic prediction model developed based on age, tumor stage, and SLC4A4 expression exhibited an efficient performance in predicting 1-, 3-, and 5-year overall survival of CRC patients. In conclusion, the current study identified several genes and pathways related to CRC, which provided new insight in understanding molecular mechanism of tumorigenesis and development of CRC.

This case-control study aims to investigate the correlations of insulin-like growth factor receptor 1 (IGF-1R) and cyclooxygenase 2 (COX-2) expressions with Ras and BRAF genetic mutations, clinicopathological features and prognosis of colorectal cancer (CRC) patients. A total of 213 CRC patients (case group) and 200 healthy individuals (control group) were selected from our hospital. Ras (K-Ras/N-Ras) and BRAF genetic mutations were detected by direct sequencing. The positive expression rates of IGF-IR and COX-2 in CRC and normal tissues were detected using immunohistochemistry. RT-qPCR and Western blotting were applied to detect the mRNA and protein expressions of IGF-IR and COX-2 in CRC tissues and normal tissues. Total mutation rate of N-Ras, BRAF and K-Ras in case group were 5.2%, 12.2% and 47.4%, respectively. The expressions of IGF-IR and COX-2 were higher in CRC tissues with Ras and BRAF mutations than in those without. CRC tissues with Ras mutation showed higher COX-2 expression than those with BRAF mutation. IGF-IR and COX-2 expressions were correlated to infiltration degree, lymphatic metastasis (in CRC tissues with and without Ras and BRAF mutations), and Dukes stages (only in CRC tissues with Ras and BRAF mutations). CRC patients with negative expressions of IGF-IR and COX-2 had significantly higher accumulative survival rate and longer mean survival duration than those with positive expressions of IGF-IR and COX-2. These findings indicate that IGF-1R and COX-2 expressions may be associated with Ras and BRAF genetic mutations, clinicopathological feature and prognosis of CRC patients.

e15557 Background: R-spondin 3 (RSPO3) is a potentiator of the Wnt/β-catenin pathway and its dysregulation has been implicated in cancer development. However, the clinicopathologic features of RSPO3 expression and targeting RSPO3 therapy in colorectal cancer (CRC) remain largely unknown. Methods: Serum RSPO3 level was measures by ELISA in CRC patients, individuals with colon polyps, and healthy controls, respectively. RSPO3 expression in CRC tissues were assessed through immunohistochemistry (IHC) on tissue arrays from 100 pairs of CRC and adjacent normal tissues. Transcription levels of RSPO3 were analyzed using Real-Time PCR in 30 pairs of CRC and non-tumor tissues. The prognostic significance of RSPO3 was evaluated in a cohort of 279 CRC patients. The correlation between RSPO3 expression and immune cell infiltration were analyzed in human CRC tissues. The RSPO3-overexpressing CRC cells lines were established and the effects of RSPO3 on CRC tumor growth was explored using subcutaneous tumor mouse models. Results: The RSPO3 levels in serum was significantly lower in CRC patients than in colon polyp patients (p &lt; 0.01) and healthy individuals (p &lt; 0.001). The mRNA expression of RSPO3 was significantly decreased in CRC tissues compared with the paired adjacent tissues in 73% (22/30) of CRC cases. Accordingly, the protein expression of RSPO3 was significantly lower in CRC tissues than in adjacent normal tissues in 70% (70/100) of CRC patients. Interestedly, the low expression of RSPO3 indicated less infiltration of CD11C+ dendritic cells (DCs), NK cells and CD8+ T cells in CRC tissues (R = 0.38, p &lt; 0.05). Kaplan-Meier survival analysis revealed that low expression of RSPO3 was correlated with poor prognosis in CRC (p &lt; 0.01). The restoration expression of RSPO3 in human (RKO and HCT116) and mouse (MC38) CRC cells did not affect the cell proliferation, cell cycle and apoptosis, but significantly inhibited tumor growth in both immunocompetent and immunocompromised mice, implicating that RSPO3 inhibits tumor growth through activation of immune response in tumor microenvironment (TME). Importantly, recombinant RSPO3 (rRSPO3) treatment significantly inhibited CRC tumor growth in xenograft and allograft mouse models. rRSPO3 and anti-PD-1 combination exhibited greater inhibitory effects than either treatment alone (p &lt; 0.05). FACS analysis confirmed that rRSPO3 treatment notably increased the infiltration of various immune cells, particularly DCs, in the TME (p &lt; 0.05). Conclusions: RSPO3 is lowly expressed in CRC and its low expression was associated with less infiltration of immune cells and poor prognosis in CRC patients. The restoration of RSPO3 expression inhibits CRC tumor growth in vivo . Our results for the first time demonstrate that RSPO3 function as a tumor suppressor that activates in vivo immune response and has a potential implication for CRC immunotherapy.

BackgroundThe B3GNT6 protein is a member of the O-GlcNAc transferase (OGT) family and is responsible for the production of the core 3 structure of O-glycans. It is generally expressed in the gastrointestinal (GI) tract; however, its clinical significance in colorectal cancer remains largely unexplored.MethodsWe obtained mRNA transcriptomic sequencing data from 3 gene expression omnibus (GEO) datasets (GSE37182, GSE39582, GSE103512) and The Cancer Genome Atlas (TCGA) to compare the B3GNT6 mRNA levels between colorectal cancer and normal tissues and further evaluate its value as a prognostic marker in colorectal cancer. We further validated this at the protein level in our cohort using immunohistochemical staining of B3GNT6 as well as the Human Protein Atlas online database.ResultsB3GNT6 expression was downregulated in colorectal cancer tissues as compared to that in the normal tissues at both mRNA and protein levels. Downregulation of B3GNT6 expression was found to be associated with poor overall survival in patients with colorectal cancer as per the data in GSE39582 and TCGA databases. Low B3GNT6 mRNA levels were significantly associated with chromosome instability (CIN) and KRAS mutations in patients with colorectal cancer. Gene set enrichment analysis (GSEA) revealed that low B3GNT6 expression levels in colorectal cancer were associated with increased proteasome activity.ConclusionsThe results of this study demonstrate that low expression of B3GNT6 is a potential biomarker for poor outcomes in patients with CRC. Moreover, the low expression of B3GNT6 may indicate more frequent activation of the KRAS/ERK signaling pathway, high CIN, and increased proteasomal activity. These novel findings may prove helpful for molecular diagnosis and provide a new therapeutic target for colorectal cancer.

Colorectal cancer (CRC) is a common tumor of the digestive system. In recent years, CRC has had the fourth highest incidence and second highest mortality rate among tumors worldwide. Therefore, it is important to identify new molecular markers, and examine their relationship with the clinicopathological features and prognosis of CRC patients. This study examined the correlation between the expression levels of the tropomyosin receptor kinase C (TRKC) and neurotrophic factor-3 (NT-3) proteins, and the clinicopathological features and prognosis of CRC patients. In total, 141 paraffin-embedded specimens from patients who had undergone radical surgical resection for CRC were analyzed. All CRC diagnoses were confirmed by pathological examination. Corresponding adjacent normal tissues served as controls. Immunohistochemical staining was employed to assess the expression of the TRKC and NT-3 proteins in both the CRC and adjacent normal tissues. A subsequent statistical analysis was conducted to explore the associations between the expression levels of these proteins, and the clinicopathological features and prognostic outcomes of CRC patients. The expression level of TRKC was significantly higher in the CRC tissues than the adjacent normal tissues, while the expression level of NT-3 was significantly lower in the CRC tissues than the adjacent normal tissues, and the observed differences were statistically significant (P<0.001). Notably, TRKC expression was significantly correlated with the tumor site (P<0.05), lymph node metastasis (P<0.05), tumor node metastasis (TNM) stage (P<0.01), serum carcinoembryonic antigen (CEA) level (P<0.01), and serum carbohydrate antigen 19-9 (CA199) level (P<0.05). The patients with positive TRKC had significantly shorter overall survival (OS) and progression-free survival (PFS) times than those with negative TRKC expression (P<0.001). Further, NT-3 expression was significantly associated with lymph node metastasis (P<0.05), distant metastasis (P<0.05), TNM stage (P<0.05), and serum CEA level (P<0.05). The patients with positive NT-3 expression had prolonged OS and PFS compared to those with negative NT-3 expression (P<0.01). The Cox proportional hazards regression model revealed that TRKC expression had a hazard ratio (HR) of 2.679 (P<0.01), while NT-3 expression had a HR of 0.433 (P<0.05). We found that the TRKC and NT-3 proteins were closely associated with the occurrence, metastasis, invasion, and tumor marker levels of CRC, and can be used as independent predictors of prognosis in patients with CRC. TRKC mainly indicates a poor prognosis in CRC, while NT-3 indicates a good prognosis.

TMEM16A is a recently identified calcium-activated chloride channel (CaCC) and its overexpression contributes to tumorigenesis and progression in several human malignancies. However, little is known about expression of TMEM16A and its clinical significance in colorectal cancer (CRC). TMEM16A mRNA expression was determined by quantitative real time-PCR (qRT-PCR) in 67 CRC tissues and 24 para-carcinoma tissues. TMEM16A protein expression was performed by immunohistochemistry in 80 CRC tissues. The correlation between TMEM16A expression and clinicopathological parameters, and known genes and proteins involved in CRC was analyzed. The results showed that TMEM16A mRNA expression was frequently detected in 51 CRC tissues (76%), whereas TMEM16A protein expression was determined at a relatively lower frequency (26%). TMEM16A mRNA expression in tumor tissues was higher than its expression in normal para-carcinoma tissues (P < 0.05). TMEM16A mRNA expression was significantly correlated with TNM stage (p = 0.039) and status of lymph node metastasis (p = 0.047). In addition, there was a strong positive correlation between TMEM16A mRNA expression and MSH2 protein. More importantly, TMEM16A protein expression was positively associated with KRAS mutation, and negatively correlated with mutant p53 protein. Logistic regression analysis demonstrated that TMEM16A mRNA expression was an important independent predictive factor of lymph node metastasis (OR = 16.38, CI: 1.91–140.27, p = 0.01). TMEM16A mRNA and protein expression was not significantly related with patient survival. Our findings provide original evidence demonstrating TMEM16A mRNA expression can be a novel predictive marker of lymph node metastasis and TMEM16A protein expression may be an important regulator of tumor proliferation and metastasis in CRC.

Tumor-infiltrating immune cells (TIICs) play essential roles in cancer development and progression. However, the association of TIICs with prognosis in colorectal cancer (CRC) patients remains elusive. Infiltration of TIICs was assessed using ssGSEA and CIBERSORT tools. The association of TIICs with prognosis was analyzed in 1,802 CRC data downloaded from the GEO (https://www.ncbi.nlm.nih.gov/geo/) and TCGA (https://portal.gdc.cancer.gov/) databases. Three populations of TIICs, including CD66b+ tumor-associated neutrophils (TANs), FoxP3+ Tregs, and CD163+ tumor-associated macrophages (TAMs) were selected for immunohistochemistry (IHC) validation analysis in 1,008 CRC biopsies, and their influence on clinical features and prognosis of CRC patients was analyzed. Prognostic models were constructed based on the training cohort (359 patients). The models were further tested and verified in testing (249 patients) and validation cohorts (400 patients). Based on ssGSEA and CIBERSORT analysis, the correlation between TIICs and CRC prognosis was inconsistent in different datasets. Moreover, the results with disease-free survival (DFS) and overall survival (OS) data in the same dataset also differed. The high abundance of TIICs found by ssGSEA or CIBERSORT tools can be used for prognostic evaluation effectively. IHC results showed that TANs, Tregs, TAMs were significantly correlated with prognosis in CRC patients and were independent prognostic factors (PDFS ≤ 0.001; POS ≤ 0.023). The prognostic predictive models were constructed based on the numbers of TANs, Tregs, TAMs (C-indexDFS&OS = 0.86; AICDFS = 448.43; AICOS = 184.30) and they were more reliable than traditional indicators for evaluating prognosis in CRC patients. Besides, TIICs may affect the response to chemotherapy. In conclusion, TIICs were correlated with clinical features and prognosis in patients with CRC and thus can be used as markers.

Colorectal cancer (CRC) is reported to be the third most common cancer and the fourth leading cause of cancer-related deaths around the world. MicroRNA-485 (miR-485) has been reported to be aberrantly expressed and play important roles in several types of human malignancy. However, the expression level, biological functions and underlying molecular mechanisms of miR-485 in CRC remain unclear. Therefore, the aim of the present study was to determine miR-485 expression levels and their clinical significance in CRC and to explore the functions and underlying mechanisms of miR-485 in this disease. In the present study, miR-485 was lowly expressed in CRC tissues and cell lines. Decreased miR-485 expression was associated with tumour size, lymph node metastasis, distant metastasis and TNM stage. Functional assays indicated that upregulation of miR-485 impaired CRC cell proliferation, invasion and induced cell apoptosis. Grb2-associated binding2 (GAB2) was identified as a direct target of miR-485 in CRC. GAB2 was upregulated in CRC tissues and was negatively correlated with the miR-485 expression level. Furthermore, GAB2 knockdown simulated the tumour-suppressing roles of miR-485 overexpression in CRC cells. Moreover, restored GAB2 expression reversed the effects of miR-485 overexpression in CRC cells. In addition, miR-485 suppressed the AKT and ERK signalling pathways in CRC by directly targeting GAB2. Collectively, these findings demonstrate that miR-485 may play tumour suppressive roles in CRC by directly targeting GAB2 and indirectly regulating AKT and ERK signalling pathways, suggesting that miR-485 may be a potential therapeutic target for patients with thisdisease.

3586 Background: Immune-related genes (IRGs) were found to be associated with the prognosis of colorectal cancer (CRC) patients. The aim of this study was to evaluate the impact of IRGs in predicting prognosis of early-stage CRC patients. Methods: According to the CIT microarray data set, 309 early-stage CRC patients were selected for generation of immune-related gene signature (IRGS). 5 independent data sets included 1587 CRC patients with complete prognostic information were divided into a training cohort (566 patients) and two validation cohorts (624 patients in validation-1 and 397 patients in meta-validation). Prognostic analysis were performed to test the predictive value of IRGS. Results: Of 309 early-stage CRC patients, a prognostic immune signature included 23 immune-related genes was constructed. In the training cohort, when considering patients with early tumor stage (I or II), IRGS significantly stratified patients into immune low- vs high-risk groups in terms of disease-free survival (HR = 5.03, 95%CI = 2.94-8.62, P &lt; 0.001). Similarly, higher IRGS was correlated with significantly worse prognosis of early-stage CRC patients in validation-1 (HR = 2.71, 95%CI = 1.44-5.08, P = 0.001) and meta-validation cohort (HR = 3.10, 95%CI = 1.60-6.00, P &lt; 0.001). When compared with Oncotype DX, we found IRGS achieved an improved survival correlation in the training cohort (mean C-index, 0.85 vs 0.65) and the validation-1 cohort (mean C-index, 0.72 vs 0.61). After integrated with clinical characteristics, IRGS remained as an independent prognostic factor after adjusting for T stage and TNM stage of tumor in multivariate analysis (HR = 2.02, 95%CI = 1.61-2.53, P &lt; 0.001). Furthermore, IRGS stratified immune low-risk group patients with adjuvant chemotherapy showed even worse disease-free survival when compared with those without adjuvant chemotherapy (HR = 5.66, 95%CI = 3.153-10.16, P &lt; 0.001 in the training cohort and HR = 3.21, 95%CI = 1.74-5.92, P &lt; 0.001 in the validation-1 cohort). IRGS identified immune high-risk group obtained a significantly higher immune and stromal infiltration (P &lt; 0.001). Particularly, the percentages of Macrophages M2 and CD8+ T cells infiltration were significantly different between these two groups. Conclusions: The proposed prognostic IRGS is a promising system for estimating DFS of colorectal cancer patients, especially those in early-stage. Further studies are needed to evaluate the clinical utility of this system in predicting prognosis of colorectal cancer patients.