Distribution and prognostic value of macrophages in colorectal cancer and adjacent mucosa in patient stages I-III vs IV

Table 1 Clinical backgrounds of enrolled patients and histopathological features of primary colorectal cancer and liver metastases, n (%)/median (minimum-maximum)

Parameter			Stage I-III (n = 44)	Stage IV (n = 55)	P value
Age at the diagnosis (years)			64 (46-73)	62 (29-78)	0.31
Gender		Male	29 (65.9)	34 (61.8)	0.67
		Female	15 (34.1)	21 (38.2)
Primary tumor
Location		Right colon	8 (18.2)	12 (21.8)	0.65
		Left colon	36 (81.8)	43 (78.2)
Size (cm)			3.5 (0.8-8.3)	4.3 (1.0-7.5)
Pathologic T stage		T1	1 (2.3)	0 (0.0)	0.25
		T2	6 (13.6)	1 (1.8)
		T3	35 (79.5)	48 (87.3)
		T4	2 (4.5)	6 (10.9)
Histological type		Not otherwise specified	39 (88.6)	51 (92.7)
		Mucinous	4 (9.1)	2 (3.6)
		Other	1 (2.3)	2 (3.6)
Pathologic N stage		N0	13 (29.5)	13 (23.6)	0.51
		N1	16 (36.4)	22 (40.0)
		N2	15 (34.1)	20 (36.4)
American Joint Committee on Cancer 8th staging		Stage I	1 (2.3)
		Stage II	12 (27.3)
		Stage III	31 (70.4)
		Stage IV		55 (100)
Grade		1	14 (31.8)	12 (21.8)	0.77
		2	26 (59.1)	37 (67.3)
		3	4 (9.1)	6 (10.9)
KRAS status		Mutated	11 (25.0)	20 (36.4)	0.26
		Wild-type	25 (56.8)	27 (49.1)
		Not tested	8 (18.2)	8 (14.5)
BRAF status		Mutated	1 (2.3)	0 (0.0)	0.27
		Wild-type	35 (79.5)	44 (80.0)
		Not tested	8 (18.2)	11 (20.0)
Microsatellite instability status		High	1 (2.3)	1 (1.8)	0.90
		Low	35 (79.5)	42 (76.4)
		Not tested	8 (2.3)	12 (21.8)
CEA ng/mL			8.9 (0.8-1649.0)	6.8 (1.0-945.0)	0.90
CEA		< 5 ng/mL	13 (46.4)	18 (46.2)
		> 5 ng/mL	15 (53.6)	21 (53.8)	0.98
Number of examined lymph nodes			11 (0-43)	13 (1-34)	0.38
Lymph node ratio			0.13 (0-0.32)	0.22 (0-0.57)	0.14
Liver metastases
Number			1 (1-7)	2 (1-24)	0.08
Size (cm)			2.7 (0.6-7.1)	2.0 (0.4-26.0)	0.02
Grade	1		18 (42.9)	21 (39.6)	0.75
	2		24 (57.1)	32 (60.4)
Metastasis resection	R0		29 (69.0)	40 (76.9)	0.39
R status	R1		13 (31.0)	12 (23.1)
Chemotherapy ± targeted therapy	CHT alone		30 (68.2)	28 (50.9)	0.07
	CHT + anti-vascular endothelial growth factor		4 (9.1)	11 (20.0)
	CHT + anti-epidermal growth factor receptor		5 (11.4)	9 (16.4)
	No treatment		2 (4.5)	4 (7.3)
	Not known		3 (6.8)	3 (5.4)
Chemotherapy regimens	FOLFOX		16 (36.4)	31 (56.4)	0.05
	Dle Gramont		9 (20.5)	7 (12.7)
	FOLFIRI		2 (4.5)	4 (7.3)
	Others		12 (27.3)	6 (10.9)

CEA: Carcinoembryonic antigen; CHT: Chemotherapy.

Table 2 Hazard ratios for overall survival between high vs low cell density of macrophages in colorectal cancer patients’ stage I-III vs IV, n (%)/hazard ratio (95%CI)

	Stages I-III		Stage IV
CD163 NM	33 (75.0)	1.09 (0.43-2.76), P = 0.89	40 (74.1)	0.45 (0.22-0.95), P = 0.04
CD163 TC	31 (73.8)	0.94 (0.39-2.30), P = 0.90	40 (75.5)	0.96 (0.47-1.93), P = 0.90
CD163 TC/NM	21 (50.0)	0.82 (0.36-1.89), P = 0.65	26 (50.0)	1.26 (0.66-2.39), P = 0.48
CD206 NM	32 (74.4)	0.90 (0.37-2.20), P = 0.81	39 (75.0)	1.30 (0.62-2.72), P = 0.48
CD206 TC	32 (76.2)	1.49 (0.50-4.41), P = 0.47	40 (74.1)	0.73 (0.36-1.46), P = 0.37
CD206 TC/NM	21 (50.0)	1.41 (0.60-3.31), P = 0.43	25 (49.0)	0.54 (0.28-1.04), P = 0.06
CD68 NM	33 (75.0)	0.79 (0.34-1.87), P = 0.60	40 (74.1)	0.65 (0.33-1.30), P = 0.22
CD68 TC	31 (75.6)	0.62 (0.26-1.51), P = 0.30	40 (75.5)	0.78 (0.39-1.57), P = 0.48
CD68 TC/NM	20 (48.8)	0.31 (0.12-0.82), P = 0.02	26 (50.0)	0.84 (0.45-1.59), P = 0.59
CD80 NM	31 (73.8)	0.24 (0.10-0.57), P = 0.001	39 (75.0)	0.61 (0.29-1.30), P = 0.20
CD80 TC	30 (73.2)	1.00 (0.40-2.48), P = 0.10	39 (75.0)	0.84 (0.41-1.74), P = 0.64
CD80 TC/NM	20 (48.8)	1.04 (0.45-2.41), P = 0.93	24 (49.0)	1.04 (0.54-2.00), P = 0.92

Densities of macrophages (per mm²) were converted into percentiles and then categorized into low (0%-24%) and high (25%-100%). Ratio of tumor center (TC)/adjacent nontumor mucosa (NM) was calculated by dividing the densities of macrophages in TC by their densities in NM. Median of TC/NM ratios were used as cutoffs to define high and low ratio groups. Hazard ratios represent the relative risk compared with low density/ratio group (hazard ratio = 1.00). The n (%) refers to patients classified as the high-density/ratio group for that specific marker in each individual region. NM: Adjacent nontumor mucosa; TC: Tumor center.