Identification of macrophage-related molecular subgroups and risk signature in colorectal cancer based on a bioinformatics analysis

Figures & data

Figure 1. Association of B cell (A), dendritic cells (B), macrophage (C), neutrophil (D), T cell CD4 (E), T cell CD8 (F) with overall survival of CRC. CRC patients were divided into two subtypes based on the median level of immune cells.

Figure 2. Characterisation of immune landscape between the low- and high-macrophage subgroups. Boxplot shows the level of ESTIMATE score (A), immune score (B), stromal score (C), and tumour purity (D) between the low- and high-macrophage subgroups. Boxplot shows significantly different immunocyte infiltration between the low- and high-macrophage subgroups (E).

Figure 3. GSVA in macrophage-associated subgroups. (A) The volcano plot depicts the differentially expressed pathways between low- and high-macrophage subgroups. (B) The heatmap plot exhibits the representative pathways between low- and high-macrophage subgroups.

Figure 4. GSEA in the low- and high-macrophage subgroups.

Figure 5. Identification of DEMRGs in CRC. (A) The volcano plot depicts the MRGs between low- and high-macrophage subgroups. (B) The volcano plot depicts the DEGs between tumour and normal groups. (C) The intersection of genes between the DEGs and MRGs.

Figure 6. Establishment of the prognostic model based on macrophage-associated subgroups. (A) Identification of prognosis-associated DEMRGs using the Univariate Cox analysis. (B-C) LASSO analysis identifies seven genes used to construct the prognostic model. (D) Survival analysis of the risk score in the TCGA dataset. (E) Distributions of the risk score, survival status, and the expression level of seven genes in the low- and high-risk groups. (F) ROC curves of the prognostic model.

Figure 7. Validation of the prognostic model. (A) Survival analysis of the risk score in the GSE39582 dataset. (B) Distributions of the risk score, survival status, and the expression level of seven genes in the low- and high-risk groups. (C) ROC curves of the prognostic model.

Figure 8. Development of the nomogram for CRC patients. (A) The overall survival nomogram plot. (B) The calibration plot of the nomogram.

Table 1. Univariate and multivariate COX analyses for risk score and clinicopathological features.

Characteristics	Total(N)	Univariate analysis		Multivariate analysis
		Hazard ratio (95% CI)	P value	Hazard ratio (95% CI)	P value
Age	454	1.024 (1.007-1.043)	0.007	1.037 (1.018-1.055)	<0.001
Sex	454
FEMALE	214	Reference
MALE	240	1.138 (0.763-1.697)	0.528
T	454
T1	11	Reference
T2	77	0.455 (0.088-2.355)	0.348	0.318 (0.060-1.675)	0.176
T3	309	1.260 (0.309-5.148)	0.747	0.538 (0.068-4.235)	0.556
T4	56	3.665 (0.856-15.690)	0.080	1.086 (0.132-8.902)	0.939
N	454
N0	267	Reference
N1	105	1.690 (1.017-2.808)	0.043	0.601 (0.201-1.803)	0.364
N2	82	4.017 (2.534-6.369)	<0.001	0.973 (0.340-2.789)	0.960
M	454
M0	333	Reference
MX	50	2.073 (1.121-3.836)	0.020	1.710 (0.879-3.326)	0.114
M1	64	4.457 (2.831-7.017)	<0.001	6.576 (2.173-19.897)	<0.001
Stage	454
Stage II	176	Reference
Stage III	128	1.815 (1.077-3.061)	0.025	2.621 (0.790-8.699)	0.115
Stage IV	64	4.788 (2.825-8.116)	<0.001
Stage I	75	0.569 (0.217-1.488)	0.250	0.933 (0.147-5.919)	0.941
Risk score	454	4.363 (2.334-8.156)	<0.001	3.444 (1.679-7.063)	<0.001

Bolded text indicates that p-values are statistically different.

Figure 9. The association between the risk score and clinicopathological features, including sex (A), age (B), N stage (C), M stage (D), T stage (E), and TNM stage (F). An asterisk (*) indicates p < 0.05, two asterisks (**) indicates p < 0.01, three asterisks (***) indicates p < 0.001, ns indicates no significant difference.

Figure 10. Characterisation of immune landscape between the low- and high-risk groups. Boxplot shows the level of ESTIMATE score (A), immune score (B), stromal score (C), and tumour purity (D) between the low- and high-risk groups. Boxplot shows significantly different immunocyte infiltration between the low- and high-risk group (E).

Figure 11. The association of risk score with immunocyte infiltration. An asterisk (*) indicates p < 0.05, two asterisks (**) indicates p < 0.01, three asterisks (***) indicates p < 0.001.

Figure 12. Validation of signature genes by qRT-PCR analysis. An asterisk (*) indicates p < 0.05, two asterisks (**) indicates p < 0.01, three asterisks (***) indicates p < 0.001.

Data availability statement

All dataset in the present study is available in TCGA (https://portal.gdc.cancer.gov/) database.