Tumor-associated macrophages remodeling EMT and predicting survival in colorectal carcinoma

Figures & data

Figure 1. Immunohistochemistry staining results. (A) The construction of tissue array (5x), scale bar: 100 μm; (B-F) The staining of immune cells markers: CD68, CD4, CD8, CD3 and CD20 (20x); (G) The staining of cancer stemness markers: membranous CD44v6 (20x), scale bar: 100 μm; (H-I) The staining of epithelial-mesenchymal transition markers: membranous E-cadherin and nuclear Snail (20x), scale bar: 100 μm.

Table 1. The correlations between density of CD68⁺ macrophages and other immune cells in TF (Spearman rank test).

		CD68^+TF
Variables		low	high	p (rho value)
CD3^+TF	low	100	68	0.002 (0.172)
	high	63	86
CD4^+TF	low	94	86	0.71
	high	92	91
CD8^+TF	low	114	75	<0.001 (0.187)
	high	69	97
CD20^+TF	low	112	82	0.004 (0.152)
	high	67	91

Table 2. Correlation between CD68⁺ macrophages and expression of CSC and EMT markers in TF (Spearman rank test).

		CD68^+TF
variables		low	high	p (rho value)
CD44v6TF	low	106	81	<0.001 (0.211)
	high	33	63
E-cadherinTF	low	33	33	0.808
	high	98	105
snailTF	low	141	144	0.08 (−0.096)
	high	28	16
Tumor bud	low	67	50	<0.001 (−0.048)
	mediate	85	122
	high	58	28

Figure 2. The survival analyses by Kaplan-Meier survival analysis (Log-rank test). (A-C) High CD20⁺_TC density, high CD68⁺_TF density and low tumor buds predicted favorable overall survival respectively (death/ total); (D) CD68⁺_TF^low TB^high had the worst survival (CD68⁺_TF^low TB^high vs CD68⁺_TF^high TB^high, p < 0.001; CD68⁺_TF^low TB^high vs CD68⁺_TF^low TB^low, p < 0.001; CD68⁺_TF^low TB^high vs CD68⁺_TF^high TB^low, p = 0.008). In high TB group, when CD68⁺_TF were high, overall survival time was prolonged, which was similar to low TB group (death/ total). ### p < 0.001.

Table 3. The results of multivariate COX proportional hazard model.

variables	β	RR (CI 95%)	p
TNM stage			<0.001
TNM stage(I)
TNM stage(II)	0.205	1.228(0.527-2.286)	0.634
TNM stage(III)	0.741	2.097(0.946-4.652)	0.068
TNM stage(IV)	1.749	5.747(2.467-13.384)	<0.001
CD68 and TB (MATB)			0.002
CD68^low TB^high
CD68^high TB^low	−0.631	0.531(0.243-1.166)	0.115
CD68^lowTB^low	−0.955	0.385(0.197-0.752)	0.005
CD68^high TB^high	−0.803	0.448(0.273-0.737)	0.002

Figure 3. The model shows a hypothesis that tumor cells with stem-like phenotypes recruitment TAMs (brown, single sword) to the invasive front of tumor lesion and surround around tumor buds (double swords). Subsequently, TAMs induce immune response, especially CD8⁺T and CD20⁺ B lymphocytes, to eliminate these highly invasive tumor buds.