Prognostic value of circulating tumor cells combined with neutrophil-lymphocyte ratio in patients with hepatocellular carcinoma

Table 1 Clinical characteristics of the hepatocellular carcinoma patients in the study

Variable	n	%
Patients	124	100.0
Sex
Male	112	90.3
Female	12	9.7
Age (years)
< 60	73	58.9
≥ 60	51	41.1
HBV
Negative	54	43.5
Positive	70	56.5
Smoke
Negative	62	50.0
Positive	62	50.0
Alcohol
Negative	71	57.3
Positive	53	42.7
Hypertension
Negative	94	75.8
Positive	30	24.2
Diabetes mellitus
Negative	107	86.3
Positive	17	13.7
Liver cirrhosis
Negative	78	62.9
Positive	46	37.1
Child-Pugh class
A	65	52.4
B	59	47.6
BCLC stage
B	9	7.3
C	115	92.7
ALBI grade
1	64	51.6
2	55	44.4
3	5	4.0
Serum AFP, ng/mL
< 400	75	60.5
≥ 400	49	39.5
Number of tumors
< 2	38	30.6
≥ 2	86	69.4
Tumor diameter, cm
< 5	49	39.5
≥ 5	75	60.5
PVTT
Negative	69	55.6
Positive	55	44.4
Lymph node metastasis
Negative	43	34.7
Positive	81	65.3
Extrahepatic metastases
Negative	85	68.5
Positive	39	31.5

HBV: Hepatitis B virus; BCLC: Barcelona clinic liver cancer; ALBI: Albumin-bilirubin; AFP: Alpha-fetoprotein; PVTT: Portal vein tumor thrombus; NLR: Neutrophil-lymphocyte ratio; CTC: Circulating tumor cell.

Table 2 The correlation between circulating tumor cell PD-L1 (+) and neutrophil-lymphocyte ratio with clinical variables

Variable	CTC PD-L1 (+) ≤ 1, n = 68	CTC PD-L1 (+) > 1, n = 56	P value	NLR ≤ 3.89, n = 67	NLR > 3.89, n = 57	P value
Sex			0.798			0.768
Male	61	51		61	51
Female	7	5		6	6
Age (yr)			0.705			0.569
< 60	39	34		41	32
≥ 60	29	22		26	25
HBV			0.218			0.765
Negative	33	21		30	24
Positive	35	35		37	33
Smoke			0.470			0.207
Negative	36	26		37	25
Positive	32	30		30	32
Alcohol			0.264			0.337
Negative	42	29		41	30
Positive	26	27		26	27
Hypertension			0.849			0.739
Negative	52	42		50	44
Positive	16	14		17	13
Diabetes mellitus			0.866			0.535
Negative	59	48		59	48
Positive	9	8		8	9
Liver cirrhosis			0.020			0.011
Negative	49	29		49	29
Positive	19	27		18	28
Child-Pugh class			0.898			0.751
A	36	29		36	29
B	32	27		31	28
BCLC stage			0.459			0.430
B	6	3		6	3
C	62	53		61	54
ALBI grade			0.214			0.612
1	38	26		37	27
2	29	26		28	27
3	1	4		2	3
Serum AFP, ng/mL			0.153			0.017
< 400	45	30		47	28
≥ 400	23	26		20	29
Number of tumors			0.472			0.549
< 2	19	19		19	19
≥ 2	49	37		48	38
Tumor diameter, cm			0.490			0.847
< 5	25	24		27	22
≥ 5	43	32		40	35
PVTT			0.432			0.038
Negative	40	29		43	26
Positive	28	27		24	31
Lymph node metastasis			0.359			0.929
Negative	26	17		23	20
Positive	42	39		44	37
Extrahepatic metastases			0.590			0.454
Negative	48	37		44	41
Positive	20	19		23	16

HBV: Hepatitis B virus; BCLC: Barcelona clinic liver cancer; ALBI: Albumin-bilirubin; AFP: Alpha-fetoprotein; PVTT: Portal vein tumor thrombus; NLR: Neutrophil-lymphocyte ratio; CTC: Circulating tumor cell.

Table 3 The correlation between circulating tumor cell-neutrophil-lymphocyte ratio score and clinical baseline characteristics in hepatocellular carcinoma patients

Variable	CTC-NLR			P value
	Score 0, n = 54	Score 1, n=27	Score 2, n = 43
Sex				0.005
Male	51	20	41
Female	3	7	2
Age (yr)				0.993
< 60	32	16	25
≥ 60	22	11	18
HBV				0.577
Negative	25	13	16
Positive	29	14	27
Smoke				0.152
Negative	28	17	17
Positive	26	10	26
Alcohol				0.022
Negative	31	21	19
Positive	23	6	24
Hypertension				0.451
Negative	42	18	34
Positive	12	9	9
Diabetes mellitus				0.811
Negative	47	24	36
Positive	7	3	7
Liver cirrhosis				0.007
Negative	39	20	19
Positive	15	7	24
Child–Pugh class				0.693
A	28	16	21
B	26	11	22
BCLC stage				0.685
B	5	2	2
C	49	25	41
ALBI grade				0.630
1	30	15	19
2	23	11	21
3	1	1	3
Serum AFP, ng/mL				0.097
< 400	38	16	21
≥ 400	16	11	22
Number of tumors				0.407
< 2	16	6	16
≥ 2	38	21	27
Tumor diameter, cm				0.915
< 5	21	10	18
≥ 5	33	17	25
PVTT				0.269
Negative	34	15	20
Positive	20	12	23
Lymph node metastasis				0.215
Negative	18	13	12
Positive	36	14	31
Extrahepatic metastases				0.493
Negative	38	16	31
Positive	16	11	12

HBV: Hepatitis B virus; BCLC: Barcelona clinic liver cancer; ALBI: Albumin-bilirubin; AFP: Alpha-fetoprotein; PVTT: Portal vein tumor thrombus; NLR: Neutrophil-lymphocyte ratio; CTC: Circulating tumor cell.

Table 4 Univariate Cox regression analysis based on overall survival

Variable	HR	Univariate Cox regression, 95%CI	P value
Male sex	0.784	0.282-2.182	0.641
Age ≥ 60 yr	0.967	0.553-1.692	0.907
HBV (positive)	1.091	0.621-1.917	0.763
Smoke (positive)	1.341	0.767-2.342	0.303
Alcohol (positive)	1.373	0.785-2.402	0.267
Hypertension (positive)	0.711	0.364-1.388	0.317
Diabetes mellitus (positive)	0.526	0.189-1.461	0.218
Liver cirrhosis (positive)	2.283	1.305-3.996	0.004
Child-Pugh class (A-B)	1.619	0.919-2.853	0.095
BCLC stage (B-C)	2.063	0.501-8.499	0.316
ALBI grade			0.026
1
2	1.589	0.889-2.842	0.118
3	4.111	1.394-12.126	0.010
AFP ≥ 400 ng/mL	2.697	1.535-4.740	0.001
Number of tumors ≥ 2	1.243	0.670-2.307	0.490
Tumor diameter ≥ 5 cm	1.016	0.576-1.793	0.957
PVTT (positive)	1.737	0.992-3.042	0.053
Lymph node metastasis (positive)	1.695	0.911-3.154	0.095
Extrahepatic metastases (positive)	1.202	0.669-2.159	0.539
CTC PD-L1 (+) > 1	6.657	3.590-12.345	< 0.001
NLR > 3.89	3.841	2.136-6.908	< 0.001
CTC-NLR			< 0.001
0
1	2.748	1.229-6.145	0.014
2	9.720	4.679-20.192	0.000

HBV: Hepatitis B virus; BCLC: Barcelona clinic liver cancer; ALBI: Albumin-bilirubin; AFP: Alpha-fetoprotein; PVTT: Portal vein tumor thrombus; NLR: Neutrophil-lymphocyte ratio; CTC: Circulating tumor cell; HR: Hazard ratio; 95%CI: 95% confidence interval.

Table 5 Multivariate Cox regression analysis based on overall survival

Variable	HR	Overall survival, 95%CI	P value
Model 1
NLR > 3.89	1.603	0.789-3.257	0.192
CTC PD-L1 (+) > 1	5.381	2.581-11.215	< 0.001
Liver cirrhosis (positive)	1.251	0.693-2.258	0.457
ALBI grade			0.347
1
2	1.149	0.631-2.095	0.649
3	2.352	0.743-7.442	0.146
AFP ≥ 400ng/mL	2.410	1.323-4.390	0.004
Model 2
CTC-NLR			< 0.001
0
1	2.684	1.187-6.070	0.018
2	8.470	3.957-18.129	0.000
Liver cirrhosis (positive)	1.261	0.697-2.282	0.442
ALBI grade			0.218
1
2	1.191	0.653-2.171	0.569
3	2.739	0.884-8.489	0.081
AFP ≥ 400ng/mL	2.096	1.180-3.723	0.012

NLR: Neutrophil-lymphocyte ratio; CTC: Circulating tumor cell; ALBI: Albumin–bilirubin; AFP: Alpha-fetoprotein; HR: Hazard ratio; 95%CI: 95% confidence interval.