The association between circulating tumor cells and Epstein-Barr virus activation in patients with nasopharyngeal carcinoma

Figures & data

Figure 1. CTCs in patients with NPC. A-D, samples of CTCs in 4 NPC patients.

Figure 2. CTM in patients with NPC. A-B, samples of CTM in 2 NPC patients.

Figure 3. CK5/6 and P63 staining by IHC. A, staining of CK5/6; B, staining of P63.

Figure 4. EBERs staining by ISH.

Table 1. Association between CTCs and TNM stage in NPC patients.

	CTC positivity
Parameters	Positive	Negative	P value^a	CTC numberMean	P value^b
T stage			0.629		0.702
T1	1(50.0%)	1(50.0%)		0.50
T2	1(50.0%)	1(50.0%)		0.50
T3	14(70.0%)	6(30.0%)		2.50
T4	6(66.7%)	3(33.3%)		1.78
N stage			0.260		0.805
N0	2(50.0%)	2(50.0%)		1.00
N1	6(60.0%)	4(40.0%)		2.10
N2	4(66.7%)	2(33.3%)		3.00
N3	10(76.9%)	3(23.1%)		1.92
M stage			0.643		0.963
M0	18(64.3%)	10(35.7%)		2.07
M1	4(80.0%)	1(20.0%)		2.00
TNM stage^c			0.518		0.978
I	1(100.0%)	0(0.0%)		1.00
II	0(0.0%)	1(100.0%)		1.00
III	7(63.6%)	4(36.4%)		2.45
IVa	2(40.0%)	3(60.0%)		1.60
Vb	8(80.0%)	2(20.0%)		2.20
VIc	4(80.0%)	1(20.0%)		2.00

^a the P values were calculated by Linear-by-Linear Association or Fisher's Exact Test;

^b the P values were calculated by Student's t-test or ANOVA analysis;

^c the TNM staging were based on AJCC 7th.

Table 2. Association between CTCs and EBV activation in NPC Patients.

	CTC
EBV parameters	Negative	Positive	P value
Total NPC
EBV DNA^a	11.29^b	11.01	0.971
VCA-IgA^c	3.88	4.86	0.016
EA-IgA^c	3.06	3.48	0.116
NPC without metastasis
EBV DNA^a	11.29	12.60	0.878
VCA-IgA ^c	3.76	4.95	0.009
EA-IgA^c	3.00	3.57	0.056
NPC with metastasis
EBV DNA^a	/	4.29	/
VCA-IgA^c	5.07	4.38	0.478
EA-IgA^c	3.68	3.00	/

All the above analyses were performed by the t-test.

^a the crude EBV DNA load was normalized by 10³ before analysis;

^b one case with crude EBV DNA load of 1.02 × 10⁷ was treated as extreme value and was deleted before analysis.

^c VCA-IgA and EA-IgA titers were log-transformed before analysis. The statistically significant results are highlighted in bold.

Table 3. Correlation between CTC numbers and EBV hematological features in NPC Patients.

	EBV DNA		VCA-IgA		EA-IgA
	Coefficient	P	Coefficient	P	Coefficient	P
Total NPC
No. of CTC	0.396	0.033	0.382	0.041	0.263	0.169
NPC without metastasis
No. of CTC	0.417	0.038	0.449	0.024	0.319	0.120

The above analyses were calculated by the partial correlation coefficients by con-trolling the age and sex. No analysis was performed for NPC with metastasis due to limited study sample. The statistically significant results are highlighted in bold.

Table 4. Association between CTCs and blood biochemical parameters.

	Total NPC			NPC without metastasis			NPC with metastasis
Parameters	Negative	Positive	P value	Negative	Positive	P value	Negative	Positive	P value
CRP	4.0 ± 4.0	3.8 ± 6.6	0.927	3.6 ± 4.0	1.6 ± 1.3	0.051	7.8	14.0 ± 11.1	0.653
WBC	7.3 ± 2.0	6.6 ± 1.3	0.237	7.4 ± 2.1	6.7 ± 1.4	0.326	6.5	6.1 ± 0.8	0.672
NE#	4.6 ± 1.4	3.9 ± 1.3	0.153	4.7 ± 1.5	4.0 ± 1.2	0.205	4.1	3.4 ± 1.9	0.753
LY#	1.9 ± 1.0	1.9 ± 0.6	0.830	2.0 ± 1.1	2.1 ± 0.6	0.871	1.2	1.0 ± 0.2	0.427
MO#	0.5 ± 0.3	0.5 ± 0.2	0.861	0.5 ± 0.3	0.5 ± 0.2	0.836	0.6	0.6 ± 0.2	0.920
EO#	0.2 ± 0.2	0.1 ± 0.1	0.584	0.1 ± 0.2	0.1 ± 0.1	0.848	0.5	0.1 ± 0.1	0.007
BA#	0.1 ± 0.1	0.0 ± 0.0	0.223	0.1 ± 0.1	0.0 ± 0.1	0.592	0.1	0.0 ± 0.0	/
HB	143.6 ± 10.7	141.1 ± 16.0	0.633	143.2 ± 11.2	142.6 ± 15.3	0.916	148	134.0 ± 19.8	0.571
RBC	4.9 ± 0.5	4.9 ± 0.6	0.717	4.8 ± 0.4	5.0 ± 0.6	0.318	5.6	4.6 ± 0.4	0.115
PLT	251.1 ± 39.5	262.8 ± 47.6	0.487	248.9 ± 41.0	259.8 ± 46.5	0.540	273	276.3 ± 57.5	0.963
TP	73.5 ± 7.0	74.1 ± 5.2	0.781	73.3 ± 7.4	73.8 ± 5.4	0.848	75.4	75.6 ± 4.5	0.973
ALB	43.4 ± 4.4	44.5 ± 2.5	0.453	43.5 ± 4.6	44.7 ± 2.6	0.390	42.3	43.7 ± 2.2	0.623
GLOB	30.1 ± 4.0	29.6 ± 4.9	0.763	29.8 ± 4.1	29.1 ± 4.6	0.672	33.1	31.9 ± 6.6	0.885
CHO	5.6 ± 1.6	5.0 ± 0.9	0.248	5.4 ± 1.7	5.0 ± 1.0	0.376	6.7	5.2 ± 0.5	0.087
TG	1.5 ± 1.4	1.2 ± 0.5	0.252	1.5 ± 1.5	1.1 ± 0.5	0.359	2.0	1.2 ± 0.3	0.092
HDL	1.3 ± 0.3	1.4 ± 0.3	0.370	1.4 ± 0.3	1.4 ± 0.3	0.663	1.0	1.5 ± 0.4	0.288
LDL	3.7 ± 1.4	3.2 ± 0.8	0.272	3.6 ± 1.4	3.2 ± 0.9	0.344	5.1	3.3 ± 0.2	0.006
PT	11.3 ± 0.6	11.3 ± 0.7	0.928	11.3 ± 0.6	11.3 ± 0.8	0.867	11.0	11.2 ± 0.5	0.711
INR	1.0 ± 0.1	1.0 ± 0.1	0.914	1.0 ± 0.1	1.0 ± 0.1	0.836	1.0	1.0 ± 0.0	0.635
APTT	24.3 ± 2.8	26.4 ± 3.1	0.070	24.1 ± 2.8	26.2 ± 3.2	0.092	26.6	27.5 ± 2.5	0.785
Fbg	3.2 ± 0.9	2.8 ± 0.7	0.237	3.1 ± 0.8	2.6 ± 0.4	0.077	4.6	4.0 ± 1.2	0.683
TT	19.3 ± 0.9	18.9 ± 1.1	0.310	19.3 ± 1.0	19.2 ± 0.9	0.712	19.3	17.4 ± 0.9	0.205

All the above analyses were performed by the t-test. The statistically significant results are highlighted in bold.