Changes in the metabolism of axillary lymph node cells as a predictor of regional metastasis in breast cancer

Table 1 Clinical and pathological characteristics of breast cancer patients

Breast cancer characteristics	n (%)
Т stage
Т1b	4 (5.8)
T1c	42 (60.9)
Т2	23 (33.3)
N stage
N0	45 (65.1)
N1	14 (20.3)
N2	8 (11.6)
N3	2 (2.9)
Grade (G)
G1	9 (13.0)
G2	34 (49.3)
G3	26 (37.7)
Lymphovascular invasion
Absent	51 (73.9)
Present	18 (26.1)
Perineural invasion
Absent	59 (85.5)
Present	10 (14.5)
Intraductal component
Absent	57 (82.6)
Present	12 (17.4)
ER status
Negative	11 (15.9)
Positive	58 (84.1)
PR status
Negative	26 (37.7)
Positive	43 (62.3)
HER2 status
Negative	61 (88.4)
Positive	8 (11.6)
Molecular biological subtypes
Luminal A	34 (49.3)
Luminal B HER2-	23 (33.3)
Luminal B HER2+	4 (5.8)
Nonluminal HER2+	4 (5.8)
TNBC	4 (5.8)

ER: Estrogen receptor; HER2: Human epidermal growth factor receptor 2; PR: Progesterone receptor; TNBC: Triple-negative breast cancer.

Table 2 Evaluation of vascular cell adhesion molecule-1 and C-X-C motif chemokine receptor 4 expression in axillary lymph nodes

	VCAM-1	CXCR4
Marker expression in the LN sinuses
The number of sinuses	0: None
	1: ≤ 15%
	2: > 15%
The intensity of marker expression	0: No expression	0: No expression
	1: Weak expression	1: Expression present
	2: Moderate expression
	3: Pronounced expression
Marker expression scores in LN sinuses	By multiplying the number of sinuses and intensity of marker expression (score range 0-6)	By multiplying the number of sinuses and intensity of marker expression (score range 0-2)
Marker expression in the LN follicles
Number of follicles	0: < 5% of the lymph node area
	1: ≥ 5% and < 30% of the lymph node area
	2: ≥ 30% of the lymph node area
The intensity of marker expression	0: No expression	0: No expression
	1: Weak expression	1: Weak expression
	2: Moderate expression	2: Moderate expression
	3: Pronounced expression	3: Pronounced expression
Marker expression scores in LN follicles	By multiplying the number of follicles and intensity of marker expression (score range 0-6)	By multiplying the number of follicles and intensity of marker expression (score range 0-6)
Marker expression in the LN cortex
The intensity of marker expression	0: No expression	0: No expression
	1: Weak expression	1: Weak expression
	2: Moderate expression	2: Moderate expression
Marker expression in the LN medulla
The intensity of marker expression	0: No expression	0: No expression
	1: Weak expression	1: Weak expression
	2: Moderate expression	2: Moderate expression
Marker expression in metastases cells
Number of cells with marker expression	1: < 50% of cells	1: < 50% of cells
	2: 50% or more of cells	2: 50% or more of cells
The intensity of marker expression	0: No expression	0: No expression
	1: Weak expression	1: Weak expression
	2: Moderate expression	2: Moderate expression
	3: Pronounced expression
Marker expression scores in metastases	By multiplying the number of cells with marker expression and intensity of marker expression (score range 0-4)	By multiplying the number of cells with marker expression and intensity of marker expression (score range 0-4)

CXCR4: C-X-C motif chemokine receptor 4; LN: Lymph node; VCAM: Vascular cell adhesion molecule-1.

Table 3 Values of the delayed luminescence of erythrosine depending on the nature of changes in the axillary lymph nodes

Normal LN	Reactive LN	Metastases	P value
			Normal LN and reactive LN	Normal LN and metastases	Reactive LN and metastases
Indicator 1
0.916 ± 0.13	0.928 ± 0.13	0.782 ± 0.13	0.2356	0.0065a	0.0035
Indicator 2
1.236 ± 1.28	1.160 ± 0.15	1.278 ± 0.32	0.0119a	0.0012a	0.0165a
Indicator 3
10.757 ± 4.67	11.522 ± 4.94	4.670 ± 1.52	0.4834	< 0.00001a	0.00003a
Indicator 4
0.867 ± 0.16	0.886 ± 0.16	0.689 ± 0.14	0.6973	0.00002a	0.00014a
Indicator 5
0.117 ± 0.21	0.105 ± 0.11	0.215 ± 0.15	0.2961	0.0005a	0.0165a
Indicator 6
18.361 ± 10.54	18.640 ± 6.06	14.491 ± 5.06	0.0715	0.0586	0.0092a

^aP value < 0.0250 is considered significant.

LN: Lymph node.

Table 4 Vascular cell adhesion molecule-1 expression scores according to the nature of the changes in axillary lymph nodes

Normal LN	Reactive LN	Metastases	P value
			Normal LN and reactive LN	Normal LN and metastases	Reactive LN and metastases
VCAM-1 expression in the LN sinuses
0.758 ± 1.05	1.600 ± 1.43	1.125 ± 1.05	0.0989	0.1467	0.310
VCAM-1 expression in the LN follicles
0.694 ± 0.84	3.000 ± 1.33	1.625 ± 1.928	< 0.0001a	0.2063	0.0604
VCAM-1 expression in the LN cortex (B zone)
0.758 ± 0.84	1.200 ± 1.03	0.500 ± 0.73	0.2226	0.2827	0.1214
VCAM-1 expression in the LN medulla (T zone)
1.000 ± 0.77	1.600 ± 0.52	1.375 ± 0.50	0.0307	0.0783	0.3630

^aP value < 0.0250 is considered significant.

LN: Lymph node; VCAM: Vascular cell adhesion molecule-1.

Table 5 C-X-C motif chemokine receptor 4 expression scores according to the nature of the changes in the axillary lymph nodes

Normal LN	Reactive LN	Metastases	P value
			Normal LN and reactive LN	Normal LN and metastases	Reactive LN and metastases
CXCR4 expression in the LN sinuses
0.696 ± 0.60	1.000 ± 0.89	0.500 ± 0.52	0.4347	0.3942	0.2908
CXCR4 expression in the LN follicles
1.107 ± 1.06	1.667 ± 1.37	2.000 ± 1.21	0.2654	0.0235a	0.8916
CXCR4 expression in the LN cortex (B zone)
0.929 ± 0.60	2.000 ± 0.89	0.833 ± 0.39	0.0079a	0.7089	0.0135a
CXCR4 expression in the LN medulla (T zone)
1.000 ± 0.60	1.000 ± 0.00	1.167 ± 0.39	1.000	0.4690	0.6164

^aP value < 0.0250 is considered significant.

CXCR4: C-X-C motif chemokine receptor 4; LN: Lymph node.

Table 6 Correlations between indicators of delayed luminescence of erythrosine and clinical and pathological characteristics of patients with breast cancer

	Spearman-R	t(N-2)	P value
Age
Indicator 1	0.128296	1.42888	0.155595
Indicator 2	-0.155614	-1.74002	0.084378
Indicator 3	0.035913	0.39693	0.692115
Indicator 4	0.074874	0.82934	0.408530
Indicator 5	-0.078022	-0.86442	0.389055
Indicator 6	-0.071288	-0.78941	0.431403
T stage
Indicator 1	0.003854	0.04257	0.966115
Indicator 2	-0.028411	-0.31394	0.754106
Indicator 3	0.082147	0.91042	0.364395
Indicator 4	-0.000439	-0.00485	0.996139
Indicator 5	0.100514	1.11586	0.266675
Indicator 6	-0.121304	-1.34981	0.179576
N stage
Indicator 1	-0.180969	-2.03243	0.044282
Indicator 2	0.248791	2.83720	0.005331a
Indicator 3	-0.302370	-3.50379	0.000642a
Indicator 4	-0.198841	-2.24102	0.026833
Indicator 5	0.292935	3.38402	0.000961a
Indicator 6	-0.027221	-0.30078	0.764095
Tumor grade (G)
Indicator 1	-0.209999	-2.37241	0.019236a
Indicator 2	0.045306	0.50094	0.617316
Indicator 3	-0.182596	-2.05132	0.042376
Indicator 4	-0.294287	-3.40111	0.000908a
Indicator 5	0.240807	2.74044	0.007057a
Indicator 6	0.077454	0.85809	0.392528
Lymphovascular invasion
Indicator 1	-0.177852	-1.99627	0.048132
Indicator 2	0.226357	2.56682	0.011472a
Indicator 3	-0.304012	-3.52475	0.000598a
Indicator 4	-0.221884	-2.51344	0.013259a
Indicator 5	0.329835	3.85910	0.000183a
Indicator 6	0.012935	0.14288	0.886619
Perineural invasion
Indicator 1	0.104154	1.15671	0.249650
Indicator 2	0.020831	0.23013	0.818373
Indicator 3	-0.135680	-1.51262	0.132962
Indicator 4	-0.057778	-0.63925	0.523857
Indicator 5	-0.057591	-0.63717	0.525206
Indicator 6	-0.007352	-0.08121	0.935408
Intraductal component
Indicator 1	0.139169	1.55228	0.123185
Indicator 2	-0.020533	-0.22684	0.820924
Indicator 3	-0.020257	-0.22380	0.823290
Indicator 4	0.041065	0.45396	0.650667
Indicator 5	-0.052474	-0.58039	0.562722
Indicator 6	-0.122060	-1.35835	0.176858
Breast cancer subtype
Indicator 1	-0.038380	-0.42423	0.672143
Indicator 2	0.023580	0.26052	0.794900
Indicator 3	0.179638	2.01698	0.045893
Indicator 4	-0.015458	-0.17076	0.864697
Indicator 5	0.122464	1.36291	0.175420
Indicator 6	0.043013	0.47554	0.635254
Estrogen receptor status
Indicator 1	0.017593	0.19436	0.846221
Indicator 2	-0.075502	-0.83634	0.404601
Indicator 3	-0.037960	-0.41958	0.675531
Indicator 4	0.088131	0.97725	0.330381
Indicator 5	-0.166486	-1.86493	0.064594
Indicator 6	-0.161730	-1.81020	0.072727
Progesterone receptor status
Indicator 1	-0.116814	-1.29915	0.196344
Indicator 2	0.089735	0.99517	0.321623
Indicator 3	-0.111222	-1.23616	0.218775
Indicator 4	-0.069049	-0.76449	0.446049
Indicator 5	0.119409	1.32842	0.186519
Indicator 6	-0.009368	-0.10348	0.917753
Human epidermal growth factor receptor 2 status
Indicator 1	0.010464	0.11558	0.908175
Indicator 2	-0.028903	-0.31938	0.749987
Indicator 3	0.054422	0.60201	0.548287
Indicator 4	0.010573	0.11679	0.907216
Indicator 5	0.146710	1.63819	0.103959
Indicator 6	-0.034464	-0.38090	0.703941
Ki67 index
Indicator 1	-0.004886	-0.05396	0.957051
Indicator 2	0.007611	0.08407	0.933141
Indicator 3	0.217706	2.46373	0.015144a
Indicator 4	0.079141	0.87689	0.382271
Indicator 5	0.066255	0.73342	0.464709
Indicator 6	0.189863	2.13596	0.034681

^aP value < 0.0250 is considered significant.

t(N-2): The value of the Student's t-test for the number of degrees of freedom n-2.

Table 7 Values of the delayed luminescence of erythrosine in accordance with the clinical and pathological characteristics of breast cancer

	Indicator 1		Indicator 2		Indicator 3		Indicator 4		Indicator 5		Indicator 6
	mean ± SD	P value	mean ± SD	P value	mean ± SD	P value	mean ± SD	P value	mean ± SD	P value	mean ± SD	P value
Age
≥ 60 years	0.86 ± 0.12	0.5941	1.21 ± 0.12	0.2750	9.89 ± 4.56	0.3414	0.81 ± 0.18	0.4134	0.16 ± 0.10	0.0877	16.68 ± 4.71	0.8697
> 60 years	0.92 ± 0.14		1.23 ± 0.34		10.29 ± 4.88		0.87 ± 0.15		0.11 ± 0.12		18.13 ± 11.22
T stage
T1	0.90 ± 0.15	0.3033	1.24 ± 0.33	0.8437	9.75 ± 4.95	0.6492	0.85 ± 0.17	0.5262	0.12 ± 0.12	0.0917	17.32 ± 5.37	0.1069
T2	0.88 ± 0.15		1.20 ± 0.10		10.24 ± 4.66		0.83 ± 0.17		0.15 ± 0.12		18.71 ± 14.02
N stage
N0-1	0.91 ± 0.14	0.1723	1.22 ± 0.29	0.0050a	10.50 ± 4.72	0.0024a	0.86 ± 0.16	0.0609	0.11 ± 0.10	0.0011a	18.44 ± 9.73	0.0262
N2-3	0.84 ± 0.17		1.27 ± 0.13		6.88 ± 4.39		0.76 ± 0.18		0.21 ± 0.15		14.38 ± 4.75
Tumor grade (G)
G1-G2	0.89 ± 0.14	0.0644	1.28 ± 0.36	0.5353	9.68 ± 5.03	0.0854	0.85 ± 0.16	0.0112a	0.12 ± 0.13	0.0233a	17.11 ± 4.62	0.6598
G3	0.87 ± 0.17		1.19 ± 0.10		9.20 ± 4.55		0.79 ± 0.17		0.16 ± 0.10		19.71 ± 13.87
Lymphovascular invasion
No	0.92 ± 0.13	0.0485	1.22 ± 0.30	0.0116a	10.64 ± 4.67	0.0011a	0.86 ± 0.16	0.0148a	0.11 ± 0.11	0.0002a	16.85 ± 4.71	0.8889
Present	0.83 ± 0.17		1.25 ± 0.12		7.41 ± 4.64		0.77 ± 0.18		0.20 ± 0.13		20.99 ± 17.17
Perineural invasion
No	0.89 ± 0.15	0.2516	1.24 ± 0.30	0.8213	10.23 ± 4.82	0.1518	0.84 ± 0.18	0.5325	0.13 ± 0.11	0.5280	17.92 ± 9.57	0.9382
Present	0.94 ± 0.13		1.20 ± 0.10		8.28 ± 4.72		0.85 ± 0.11		0.11 ± 0.17		17.12 ± 7.30
Intraductal component
No	0.94 ± 0.13	0.1241	1.19 ± 0.12	0.8235	9.69 ± 4.91	0.8333	0.84 ± 0.16	0.6577	0.11 ± 0.16	0.5651	15.95 ± 6.25	0.1780
Present	0.89 ± 0.30		1.22 ± 0.30		9.97 ± 4.84		0.84 ± 0.17		0.13 ± 0.11		18.23 ± 9.77
Breast cancer subtype
Luminal A	0.89 ± 0.16	0.5386	1.20 ± 0.12	0.2972	9.29 ± 5.09	0.9772	0.84 ± 0.17	0.5558	0.13 ± 0.12	0.13 ± 0.12	18.19 ± 11.95
Luminal B HER2-	0.89 ± 0.11		1.30 ± 0.47		10.50 ± 4.72		0.87 ± 0.16		0.11 ± 0.12		17.18 ± 3.11	0.2159
Luminal B HER2+	0.86 ± 0.11		1.25 ± 0.03		10.00 ± 5.77		0.85 ± 0.17		0.17 ± 0.01		24.75 ± 8.62
HER2+	0.95 ± 0.14		1.12 ± 0.11		10.63 ± 4.71		0.89 ± 0.11		0.14 ± 0.13		15.16 ± 5.26
TNBC	0.90 ± 0.20		1.23 ± 0.08		11.35 ± 3.28		0.74 ± 0.21		0.20 ± 0.07		16.64 ± 3.50
Estrogen receptor status
Negative	0.93 ± 0.18	0.9020	1.18 ± 0.10	0.6652	11.03 ± 3.87	0.2022	0.81 ± 0.18	0.2178	0.17 ± 0.10	0.0554	15.98 ± 4.29	0.5662
Positive	0.89 ± 0.14		1.24 ± 0.29		9.73 ± 4.97		0.85 ± 0.16		0.12 ± 0.12		18.09 ± 9.79
Progesterone receptor status
Negative	0.92 ± 0.17	0.6149	1.20 ± 0.10	0.8460	11.04 ± 4.71	0.0848	0.85 ± 0.18	0.4543	0.11 ± 0.13	0.4510	18.06 ± 5.33	0.2230
Positive	0.89 ± 0.13		1.24 ± 0.32		9.38 ± 4.83		0.84 ± 0.16		0.14 ± 0.11		17.66 ± 10.60
Human epidermal growth factor receptor 2 status
Negative	0.89 ± 0.15	0.8055	1.24 ± 0.29	0.4746	9.86 ± 4.86	0.5958	0.84 ± 0.17	0.7991	0.13 ± 0.12	0.4746	17.73 ± 9.39	0.6171
Positive	0.93 ± 0.14		1.17 ± 0.11		10.42 ± 4.83		0.88 ± 0.12		0.15 ± 0.10		18.36 ± 7.76
Ki67 index
< 40%	0.90 ± 0.15	0.7704	1.19 ± 1.12	0.0561	9.31 ± 4.99	0.0315	0.84 ± 0.16	0.2328	0.12 ± 0.12	0.02349a	17.88 ± 10.88	0.6379
≥ 40%	0.90 ± 0.14		1.31 ± 0.43		11.11 ± 4.32		0.85 ± 0.18		0.15 ± 0.13		17.61 ± 4.56

^aP value < 0.0250 is considered significant.

HER2: Human epidermal growth factor receptor 2; TNBC: Triple-negative breast cancer.

Table 8 Correlations of vascular cell adhesion molecule-1 expression with the clinical and pathological characteristics of patients with breast cancer

VCAM-1 expression	Spearman-R	t(N-2)	P value
Age
VCAM-1 expression in the LN sinuses	-0.098419	-0.91715	0.361627
VCAM-1 expression in the LN follicles	0.384898	3.86734	0.000213a
VCAM-1 expression in the LN cortex	-0.124969	-1.16807	0.246005
VCAM-1 expression in the LN medulla	0.116966	1.09220	0.277798
T stage
VCAM-1 expression in the LN sinuses	-0.128938	-1.20579	0.231207
VCAM-1 expression in the LN follicles	-0.051366	-0.47698	0.634587
VCAM-1 expression in the LN cortex	-0.042704	-0.39638	0.692807
VCAM-1 expression in the LN medulla	-0.096888	-0.90275	0.369180
N stage
VCAM-1 expression in the LN sinuses	-0.003196	-0.02964	0.976420
VCAM-1 expression in the LN follicles	0.281316	2.71861	0.007929a
VCAM-1 expression in the LN cortex	-0.109254	-1.01928	0.310928
VCAM-1 expression in the LN medulla	0.099671	0.92894	0.355523
Tumor grade (G)
VCAM-1 expression in the LN sinuses	0.280088	2.70573	0.008217a
VCAM-1 expression in the LN follicles	0.417462	4.26037	0.000052a
VCAM-1 expression in the LN cortex	0.254710	2.44265	0.016626a
VCAM-1 expression in the LN medulla	0.220123	2.09267	0.039326
Lymphovascular invasion
VCAM-1 expression in the LN sinuses	-0.226550	-2.15703	0.033794
VCAM-1 expression in the LN follicles	0.121831	1.13829	0.258161
VCAM-1 expression in the LN cortex	-0.305464	-2.97495	0.003803a
VCAM-1 expression in the LN medulla	-0.097202	-0.90571	0.367621
Perineural invasion
VCAM-1 expression in the LN sinuses	0.104121	0.97086	0.334342
VCAM-1 expression in the LN follicles	0.034247	0.31778	0.751422
VCAM-1 expression in the LN cortex	-0.028020	-0.25995	0.795526
VCAM-1 expression in the LN medulla	-0.110634	-1.03232	0.304818
Intraductal component
VCAM-1 expression in the LN sinuses	0.069913	0.64993	0.517468
VCAM-1 expression in the LN follicles	-0.079382	-0.73849	0.462225
VCAM-1 expression in the LN cortex	-0.192815	-1.82228	0.071888
VCAM-1 expression in the LN medulla	-0.265459	-2.55337	0.012432a
Breast cancer subtype
VCAM-1 expression in the LN sinuses	0.046432	0.43106	0.667503
VCAM-1 expression in the LN follicles	0.287880	2.78770	0.006533a
VCAM-1 expression in the LN cortex	0.191185	1.80629	0.074370
VCAM-1 expression in the LN medulla	0.211896	2.01071	0.047489
Estrogen receptor status
VCAM-1 expression in the LN sinuses	-0.231564	-2.20743	0.029946
VCAM-1 expression in the LN follicles	-0.492942	-5.25405	0.000001a
VCAM-1 expression in the LN cortex	-0.358117	-3.55694	0.000613a
VCAM-1 expression in the LN medulla	-0.231909	-2.21091	0.029696
Progesterone receptor status
VCAM-1 expression in the LN sinuses	0.011830	0.10972	0.912888
VCAM-1 expression in the LN follicles	-0.081581	-0.75908	0.449882
VCAM-1 expression in the LN cortex	0.052052	0.48337	0.630065
VCAM-1 expression in the LN medulla	0.180134	1.69827	0.093072
Human epidermal growth factor receptor 2 status
VCAM-1 expression in the LN sinuses	0.377368	3.77897	0.000290a
VCAM-1 expression in the LN follicles	0.548298	6.08013	0.000000a
VCAM-1 expression in the LN cortex	0.505797	5.43738	0.000000a
VCAM-1 expression in the LN medulla	0.360459	3.58367	0.000561a
Ki67 index
VCAM-1 expression in the LN sinuses	0.183432	1.73044	0.087138
VCAM-1 expression in the LN follicles	0.427231	4.38203	0.000033a
VCAM-1 expression in the LN cortex	0.321769	3.15156	0.002235a
VCAM-1 expression in the LN medulla	0.355558	3.52784	0.000675a

^aP value < 0.0250 is considered significant.

t(N-2): The value of the Student’s t-test for the number of degrees of freedom n-2. LN: Lymph node; VCAM-1: Vascular cell adhesion molecule-1.

Table 9 Frequency of weak and pronounced vascular cell adhesion molecule-1 expression in lymph nodes according to the clinical and pathological characteristics of patients with breast cancer, n (%)

	VCAM-1 expression in the LN sinuses			VCAM-1 expression in the LN follicles			VCAM-1 expression in the LN cortex			VCAM-1 expression in the LN medulla
	0-1	2-3	P value	0-1	2-3	P value	0	1-2	P value	0-1	2	P value
Age
≥ 60 years	18 (56.3)	14 (43.7)	0.3383	28 (87.5)	4 (12.5)	0.0020a	12 (37.5)	20 (62.5)	0.0425	20 (62.5)	12 (37.5)	0.2100
> 60 years	40 (71.4)	16 (28.6)		31 (55.4)	25 (44.6)		33 (58.9)	23 (41.1)		38 (67.9)	18 (32.1)
T stage
T1	32 (59.3)	22 (40.7)	0.0972	37 (68.5)	17 (31.5)	0.7111	25 (46.3)	29 (53.7)	0.2523	36 (66.7)	18 (33.3)	0.8501
T2	26 (76.5)	8 (23.5)		22 (64.7)	12 (35.3)		20 (58.8)	14 (41.2)		22 (64.7)	12 (35.3)
N stage
N0-1	40 (64.5)	22 (35.5)	0.6703	41 (66.1)	21 (33.9)	0.7776	29 (46.8)	33 (53.2)	0.2062	40 (64.5)	22 (35.5)	0.6703
N2-3	18 (69.2)	8 (30.8)		18 (69.2)	8 (30.8)		16 (61.5)	10 (38.5)		18 (69.2)	8 (30.8)
Tumor grade (G)
G1-G2	46 (76.7)	14 (23.3)	0.0005a	47 (78.3)	13 (21.7)	0.0021a	37 (61.7)	23 (38.3)	0.0052a	46 (76.7)	14 (23.3)	0.0005a
G3	12 (42.9)	16 (57.1)		12 (42.9)	16 (57.1)		8 (28.6)	20 (71.4)		12 (42.9)	16 (54.1)
Lymphovascular invasion
No	32 (59.3)	22 (40.7)	0.0972	35 (64.8)	19 (35.2)	0.5748	23 (42.59)	31 (57.4)	0.0433	32 (59.3)	22 (40.7)	0.0972
Present	26 (76.5)	8 (23.5)		24 (70.6)	10 (29.4)		22 (64.7)	12 (35.3)		26 (76.5)	8 (23.5)
Perineural invasion
No	44 (64.7)	24 (35.3)	0.6606	47 (69.1)	21 (30.9)	0.5557	35 (51.5)	33 (48.5)	0.9079	42 (61.8)	26 (38.2)	0.1305
Present	14 (70)	6 (30)		12 (60.0)	8 (40)		10 (50)	10 (50)		16 (80)	4 (20)
Intraductal component
No	48 (66.7)	24 (33.3)	0.7505	47 (65.3)	25 (34.7)	0.4543	35 (48.6)	37 (51.4)	0.3148	42 (58.3)	30 (41.7)	0.0015a
Present	10 (62.5)	6 (37.5)		12 (75)	4 (25)		10 (62.5)	6 (37.5)		16 (100)	0 (0)
Breast cancer subtype
Luminal A	26 (65)	14 (35)	0.0003a	31 (77.5)	9 (22.5)	< 0.00001a	23 (57.5)	17 (42.5)	0.0008a	28 (70)	12 (30)	0.00001a
Luminal B HER2 –	22 (73.3)	8 (26.7)		22 (73.3)	8 (26.7)		16 (53.3)	14 (46.7)		24 (80)	6 (20)
Luminal B HER2 +	0 (0)	8 (100)		0 (0)	8 (100)		0 (0)	8 (100)		0 (0)	8 (100)
HER2 +	4 (100)	0 (0)		0 (0)	4 (100)		0 (0)	4 (100)		0 (0)	4 (100)
TNBC	6 (100)	0 (0)		6 (100)	0 (0)		6 (100)	0 (0)		6 (100)	0 (0)
Estrogen receptor status
Negative	10 (100)	0 (0)	0.0157a	6 (60)	4 (40)	0.6147	6 (60)	4 (40)	0.5515	6 (60)	4 (40)	0.6754
Positive	48 (61.5)	30 (38.5)		53 (67.9)	25 (32.1)		39 (50)	39 (50)		52 (66.7)	26 (33.3)
Progesterone receptor status
Negative	24 (70.6)	10 (29.4)	0.4625	18 (52.9)	16 (47.1)	0.0255	16 (47.1)	18 (52.9)	0.5437	22 (64.7)	12 (35.3)	0.8501
Positive	34 (62.9)	20 (37.1)		41 (75.9)	13 (24.1)		29 (53.7)	25 (46.3)		36 (66.7)	18 (33.3)
Human epidermal growth factor receptor 2 status
Negative	54 (71.1)	22 (28.9)	0.0104a	59 (77.6)	17 (22.4)	< 0.00001a	45 (59.2)	31 (40.8)	0.0001a	58 (76.3)	18 (23.7)	< 0.00001a
Positive	4 (33.3)	8 (66.7)		0 (0)	12 (100)		0 (0)	12 (100)		0 (0)	12 (100)
Ki67 index
< 40	38 (67.9)	18 (34.1)	0.6101	43 (76.8)	13 (23.2)	0.0101a	31 (55.4)	25 (44.6)	0.2947	44 (78.6)	12 (21.4)	0.0009a
≥ 40	20 (62.5)	12 (37.5)		16 (50)	16 (50)		14 (43.7)	18 (56.3)		14 (43.7)	18 (56.3)

^aP value < 0.0250 is considered significant.

CXCR4: C-X-C motif chemokine receptor 4; LN: Lymph node; HER2: Human epidermal growth factor receptor 2; TNBC: Triple-negative breast cancer; VCAM-1: Vascular cell adhesion molecule-1.

Table 10 Correlations of C-X-C motif chemokine receptor 4 expression with the clinical and pathological characteristics of patients with breast cancer

	Spearman-R	t(N-2)	P value
Age
CXCR4 expression in the LN sinuses	-0.033856	-0.28744	0.77460
CXCR4 expression in the LN follicles	0.186206	1.60813	0.11218
CXCR4 expression in the LN cortex	0.264398	2.32627	0.02282a
CXCR4 expression in the LN medulla	0.152145	1.30620	0.19564
T stage
CXCR4 expression in the LN sinuses	-0.363676	-3.31274	0.00145a
CXCR4 expression in the LN follicles	-0.201267	-1.74349	0.08552
CXCR4 expression in the LN cortex	-0.242079	-2.11708	0.03771
CXCR4 expression in the LN medulla	-0.026276	-0.22303	0.82414
N stage
CXCR4 expression in the LN sinuses	0.109123	0.93150	0.35471
CXCR4 expression in the LN follicles	0.587228	6.15600	<0.00001a
CXCR4 expression in the LN cortex	-0.045038	-0.38255	0.70318
CXCR4 expression in the LN medulla	0.412394	3.84111	0.00026a
Tumor grade (G)
CXCR4 expression in the LN sinuses	0.210901	1.83073	0.07128
CXCR4 expression in the LN follicles	-0.037919	-0.32199	0.74840
CXCR4 expression in the LN cortex	0.356227	3.23489	0.00184a
CXCR4 expression in the LN medulla	-0.063014	-0.53575	0.59378
Lymphovascular invasion
CXCR4 expression in the LN sinuses	0.007235	0.06139	0.95122
CXCR4 expression in the LN follicles	0.295087	2.62059	0.01070a
CXCR4 expression in the LN cortex	-0.060831	-0.51712	0.606655
CXCR4 expression in the LN medulla	0.467999	4.49357	0.00003a
Perineural invasion
CXCR4 expression in the LN sinuses	0.082836	0.70531	0.48289
CXCR4 expression in the LN follicles	-0.187479	-1.61953	0.10971
CXCR4 expression in the LN cortex	-0.195884	-1.69497	0.09440
CXCR4 expression in the LN medulla	0.095575	0.81471	0.41792
Intraductal component
CXCR4 expression in the LN sinuses	0.003855	0.03271	0.97400
CXCR4 expression in the LN follicles	-0.242443	-2.12046	0.03741
CXCR4 expression in the LN cortex	-0.226872	-1.97661	0.05192
CXCR4 expression in the LN medulla	0.115281	0.98475	0.32804
Breast cancer subtype
CXCR4 expression in the LN sinuses	0.189883	1.64107	0.10515
CXCR4 expression in the LN follicles	-0.010422	-0.08844	0.92978
CXCR4 expression in the LN cortex	0.360217	3.27649	0.00162a
CXCR4 expression in the LN medulla	0.028502	0.24195	0.80951
Estrogen receptor status
CXCR4 expression in the LN sinuses	-0.027475	-0.23322	0.81625
CXCR4 expression in the LN follicles	-0.087522	-0.74551	0.45839
CXCR4 expression in the LN cortex	0.050522	0.42924	0.66903
CXCR4 expression in the LN medulla	0.170558	1.46875	0.14626
Progesterone receptor status
CXCR4 expression in the LN sinuses	0.144467	1.23884	0.21943
CXCR4 expression in the LN follicles	0.303606	2.70380	0.00855a
CXCR4 expression in the LN cortex	0.171745	1.47929	0.14343
CXCR4 expression in the LN medulla	0.399572	3.69856	0.00042a
Human epidermal growth factor receptor 2 status
CXCR4 expression in the LN sinuses	0.158267	1.36008	0.17805
CXCR4 expression in the LN follicles	0.260292	2.28750	0.02511a
CXCR4 expression in the LN cortex	0.122659	1.04871	0.29782
CXCR4 expression in the LN medulla	0.249569	2.18686	0.03200a
Ki67 index
CXCR4 expression in the LN sinuses	0.186882	1.61418	0.11086
CXCR4 expression in the LN follicles	0.084581	0.72027	0.47369
CXCR4 expression in the LN cortex	0.397658	3.67751	0.00045a
CXCR4 expression in the LN medulla	0.159837	1.37392	0.17373

^aP value < 0.0250 is considered significant.

t(N-2): The value of the Student’s t-test for the number of degrees of freedom n-2. CXCR4: C-X-C motif chemokine receptor 4; LN: Lymph node.

Table 11 Frequency of weak and strong C-X-C motif chemokine receptor 4 expression in axillary lymph nodes according to the clinical and pathological characteristics of patients with breast cancer, n (%)

	CXCR4 expression in the LN sinuses			CXCR4 expression in the LN follicles			CXCR4 expression in the LN cortex			CXCR4 expression in the LN medulla
	0	1-2	P value	0-1	2-4	P value	0	1-3	P value	0	1-2	P value
Age
≥ 60 years	13 (46.5)	15 (53.5)	0.3208	14 (50)	14 (50)	0.8548	12 (42.8)	16 (57.2)	0.0001a	8 (28.6)	20 (71.4)	0.0188a
> 60 years	16 (34.8)	30 (65.2)		22 (47.8)	24 (52.2)		2 (4.3)	44 (95.6)		2 (4.4)	44 (95.6)
T stage
T1	12 (27.3)	32 (72.7)	0.0110a	16 (36.4)	28 (63.6)	0.0104a	8 (18.2)	36 (81.8)	0.8446	2 (4.5)	42 (95.5)	0.0063a
T2	17 (56.7)	13 (43.3)		20 (66.7)	13 (33.3)		6 (20)	24 (80)		8 (26.7)	22 (73.3)
N stage
N0-1	21 (38.9)	33 (61.1)	0.9307	34 (62.9)	20 (37.1)	0.00005a	10 (18.5)	44 (81.5)	0.8851	10 (18.5)	44 (81.8)	0.0385
N2-3	8 (40.0)	12 (60.0)		2 (10)	18 (90)		4 (20)	16 (80)		0 (0)	20 (100)
Tumor grade (G)
G1-G2	21 (42)	29 (58)	0.4536	22 (45.8)	26 (54.2)	0.7269	12 (24)	38 (76)	0.1108	6 (12)	44 (88)	0.5817
G3	8 (33.3)	16 (66.7)		12 (50)	12 (50)		2 (8.3)	22 (91.7)		4 (16.7)	20 (83.3)
Lymphovascular invasion
No	19 (43.2)	25 (56.8)	0.3942	28 (63.6)	16 (36.4)	0.0018a	8 (18.2)	36 (81.8)	0.8446	8 (18.2)	36 (81.8)	0.1549
Present	10 (33.3)	20 (66.7)		8 (26.7)	22 (73.3)		6 (20)	24 (80.0)		2 (6.7)	28 (93.3)
Perineural invasion
No	25 (43.1)	33 (59.1)	0.1891	26 (44.8)	32 (55.2)	0.2105	10 (17.2)	48 (82.8)	0.4830	8 (13.8)	50 (86.2)	0.8934
Present	4 (25.0)	12 (75.0)		10 (62.5)	6 (37.6)		4 (25)	12 (75)		2 (12.5)	14 (87.5)
Intraductal component
No	25 (40.3)	37 (59.7)	0.6499	28 (45.2)	34 (54.8)	0.1725	10 (16.1)	52 (83.9)	0.1637	8 (12.9)	54 (87.1)	0.7271
Present	4 (33.3)	8 (66.7)		8 (66.7)	4 (33.3)		4 (33.3)	8 (66.7)		2 (16.7)	10 (83.3)
Breast cancer subtype
Luminal A	12 (40)	18 (60)	0.0621	12 (40.0)	18 (60)	0.1349	10 (33.3)	20 (66.7)	0.0462	6 (20)	24 (80)	0.2354
Luminal B HER2–	16 (53.3)	14 (46.7)		18 (60.0)	12 (40)		3 (6.7)	28 (93.3)		2 (6.67)	28 (93.3)
Luminal B HER2+	0 (0)	4 (100)		0 (0)	4 (100)		0 (0)	4 (100)		0 (0)	4 (100)
HER2+	0 (0)	4 (100)		2 (50)	2 (50)		0 (0)	4 (100)		0 (0)	4 (100)
TNBC	1 (16.7)	5 (83.3)		4 (66.7)	2 (33.3)		2 (33.3)	4 (66.7)		2 (33.3)	4 (66.7)
Estrogen receptor status
Negative	1 (10)	9 (90)	0.0420	6 (60)	4 (40)	0.4399	2 (20)	8 (80)	0.9252	2 (20)	8 (80)	0.5188
Positive	28 (43.8)	36 (56.2)		30 (46.9)	34 (51.1)		12 (18.7)	52 (81.3)		8 (12.5)	56 (87.5)
Progesterone receptor status
Negative	13 (43.3)	17 (56.7)	0.5465	20 (66.7)	10 (33.3)	0.0105a	8 (26.7)	22 (73.3)	0.1599	6 (20)	24 (80)	0.1778
Positive	16 (36.4)	28 (63.6)		16 (36.4)	28 (63.6)		6 (13.6)	38 (86.4)		4 (9.1)	40 (90.9)
Human epidermal growth factor receptor 2 status
Negative	23 (35.9)	41 (64.1)	0.0162a	34 (51.5)	32 (48.5)	0.1565	14 (21.2)	52 (78.8)	0.1480	10 (15.2)	56 (84.8)	0.2365
Positive	6 (60)	4 (40)		2 (25)	6 (75)		0 (0)	8 (100)		0 (0)	8 (100)
Ki67 index
< 40	16 (34.8)	30 (65.2)	0.3196	20 (43.5)	26 (56.5)	0.2541	12 (26.1)	34 (73.9)	0.0436	8 (17.4)	38 (82.6)	0.2111
≥ 40	13 (46.4)	15 (53.6)		16 (57.1)	12 (42.8)		2 (7.1)	26 (92.9)		2 (7.1)	26 (92.9)

^aP value < 0.0250 is considered significant.

CXCR4: C-X-C motif chemokine receptor 4; LN: Lymph node; HER2: Human epidermal growth factor receptor 2; TNBC: Triple-negative breast cancer.

Table 12 Correlations of indicators of delayed luminescence of erythrosine with vascular cell adhesion molecule-1 expression

	Spearman-R	t(N-2)	P value
Indicator 1
VCAM-1 expression in the LN sinuses	-0.004898	-0.03731	0.970370
VCAM-1 expression in the LN follicles	-0.292381	-2.32846	0.023395a
VCAM-1 expression in the LN cortex	0.000602	0.00459	0.996355
VCAM-1 expression in the LN medulla	-0.035308	-0.26906	0.788836
Indicator 2
VCAM-1 expression in the LN sinuses	0.111159	0.85184	0.397804
VCAM-1 expression in the LN follicles	0.286562	2.27792	0.026433
VCAM-1 expression in the LN cortex	0.273070	2.16180	0.034773
VCAM-1 expression in the LN medulla	0.302935	2.42084	0.018636a
Indicator 3
VCAM-1 expression in the LN sinuses	-0.072530	-0.55383	0.581826
VCAM-1 expression in the LN follicles	-0.281078	-2.23055	0.029594
VCAM-1 expression in the LN cortex	0.081563	0.62324	0.535569
VCAM-1 expression in the LN medulla	-0.061304	-0.46776	0.641712
Indicator 4
VCAM-1 expression in the LN sinuses	-0.075682	-0.57803	0.565481
VCAM-1 expression in the LN follicles	-0.119127	-0.91375	0.364629
VCAM-1 expression in the LN cortex	0.105857	0.81074	0.420830
VCAM-1 expression in the LN medulla	0.096592	0.73908	0.462840
Indicator 5
VCAM-1 expression in the LN sinuses	0.111400	0.85371	0.396778
VCAM-1 expression in the LN follicles	0.461719	3.96420	0.000205a
VCAM-1 expression in the LN cortex	0.148613	1.14451	0.257113
VCAM-1 expression in the LN medulla	0.263704	2.08201	0.041764
Indicator 6
VCAM-1 expression in the LN sinuses	0.011209	0.08537	0.932260
VCAM-1 expression in the LN follicles	0.270786	2.14228	0.036381
VCAM-1 expression in the LN cortex	0.176745	1.36758	0.176719
VCAM-1 expression in the LN medulla	0.037024	0.28216	0.778825

^aP value < 0.0250 is considered significant.

t(N-2): The value of the Student’s t-test for the number of degrees of freedom n-2. LN: Lymph node; VCAM-1: Vascular cell adhesion molecule-1.

Table 13 Correlations of indicators of delayed luminescence of erythrosine with C-X-C motif chemokine receptor 4 expression

	Spearman-R	t(N-2)	P value
Indicator 1
CXCR4 expression in the LN sinuses	-0.053527	-0.37903	0.706268
CXCR4 expression in the LN follicles	-0.221404	-1.60540	0.114703
CXCR4 expression in the LN cortex	-0.103418	-0.73522	0.465643
CXCR4 expression in the LN medulla	-0.019459	-0.13762	0.891092
Indicator 2
CXCR4 expression in the LN sinuses	-0.212379	-1.53681	0.130645
CXCR4 expression in the LN follicles	0.255859	1.87149	0.067134
CXCR4 expression in the LN cortex	-0.234264	-1.70391	0.094605
CXCR4 expression in the LN medulla	0.207313	1.49848	0.140297
Indicator 3
CXCR4 expression in the LN sinuses	-0.072580	-0.51458	0.609117
CXCR4 expression in the LN follicles	-0.245256	-1.78886	0.079696
CXCR4 expression in the LN cortex	-0.082620	-0.58621	0.560369
CXCR4 expression in the LN medulla	-0.139046	-0.99285	0.325566
Indicator 4
CXCR4 expression in the LN sinuses	-0.099432	-0.70659	0.483101
CXCR4 expression in the LN follicles	-0.055230	-0.39113	0.697363
CXCR4 expression in the LN cortex	-0.035307	-0.24981	0.803757
CXCR4 expression in the LN medulla	0.103639	0.73681	0.464684
Indicator 5
CXCR4 expression in the LN sinuses	0.003261	0.02306	0.981692
CXCR4 expression in the LN follicles	0.535508	4.48369	0.000043a
CXCR4 expression in the LN cortex	-0.016864	-0.11926	0.905546
CXCR4 expression in the LN medulla	0.137211	0.97949	0.332054
Indicator 6
CXCR4 expression in the LN sinuses	0.129308	0.92209	0.360912
CXCR4 expression in the LN follicles	0.120409	0.85766	0.395171
CXCR4 expression in the LN cortex	0.057093	0.40437	0.687667
CXCR4 expression in the LN medulla	0.053138	0.37627	0.708306

^aP value < 0.0250 is considered significant.

t(N-2): The value of the Student’s t-test for the number of degrees of freedom n-2. CXCR4: C-X-C motif chemokine receptor 4; LN: Lymph node.

Table 14 Correlations between vascular cell adhesion molecule-1 and C-X-C motif chemokine receptor 4 expression

	Spearman-R	t(N-2)	P value
VCAM-1 expression in the LN sinuses
CXCR4 expression in the LN sinuses	0.277417	2.41586	0.018311a
CXCR4 expression in the LN follicles	-0.001542	-0.01290	0.989746
CXCR4 expression in the LN cortex	0.106388	0.89519	0.373755
CXCR4 expression in the LN medulla	-0.084854	-0.71251	0.478517
VCAM-1 expression in the LN follicles
CXCR4 expression in the LN sinuses	-0.018810	-0.15740	0.875381
CXCR4 expression in the LN follicles	0.409201	3.75215	0.000358a
CXCR4 expression in the LN cortex	0.138621	1.17109	0.245532
CXCR4 expression in the LN medulla	0.106283	0.89429	0.374231
VCAM-1 expression in the LN cortex
CXCR4 expression in the LN sinuses	0.128204	1.08155	0.283163
CXCR4 expression in the LN follicles	0.094375	0.79314	0.430376
CXCR4 expression in the LN cortex	0.071568	0.60032	0.550229
CXCR4 expression in the LN medulla	-0.048490	-0.40617	0.685854
VCAM-1 expression in the LN medulla
CXCR4 expression in the LN sinuses	0.072876	0.61135	0.542946
CXCR4 expression in the LN follicles	0.283586	2.47423	0.015779a
CXCR4 expression in the LN cortex	0.248515	2.14657	0.035295
CXCR4 expression in the LN medulla	0.253763	2.19498	0.031484

^aP value < 0.0250 is considered significant.

t(N-2): The value of the Student’s t-test for the number of degrees of freedom n-2. CXCR4: C-X-C motif chemokine receptor 4; LN: Lymph node; VCAM-1: Vascular cell adhesion molecule-1.