Multi-analyte analysis of cytokines that predict outcomes in patients with hepatocellular carcinoma treated with radiotherapy

Table 1 Patient, tumor and treatment characteristics

Characteristic		n	Percent
Age (yr), median	54 (range, 32-75)
Sex	Male	44	86.3
	Female	7	13.7
Viral type	B	44	86.3
	C	4	7.8
	NBNC	3	5.9
Child-Pugh class	A	49	96.1
	B	2	3.9
Modified UICC stage	II	4	7.8
	III	32	62.7
	IV	15	29.5
BCLC stage	A	14	27.5
	B	12	23.5
	C	25	49.0
PVT	No	31	60.8
	Yes	20	39.2
Multiplicity	No	25	49.0
	Yes	26	51.0
Tumor size in cm, median		8.5 (range, 1.0-19.6)
AFP (IU/mL), median	141.9 (range, 1.53-83000)
PIVKA-II (mAU/mL), median	600 (range, 12-2000)
Treatment-naïve	Yes	29	56.9
	No	22	43.1
Total dose of RT (Gy), median		50.4 (range, 45-64.8)

NBNC: Non-B non-C; UICC: International Union Against Cancer; BCLC: Barcelona Clinic Liver Cancer; PVT: Portal vein thrombosis; AFP: α-fetoprotein; PIVKA-II: Prothrombin induced by vitamin K absence-II; RT: Radiotherapy.

Table 2 Univariate and multivariate analyses of clinical factors or cytokines for failure-free survival - overall survival

	Univariate		Multivariate
	P value	RR (95%CI)	P value	RR (95%CI)
Sex	0.580	1.276 (0.538-3.027)
Age	0.954	1.001 (0.970-1.033)
Stage	0.861	1.052 (0.600-1.844)
BCLC	0.224	1.252 (0.872-1.799)
Size	0.308	1.037 (0.967-1.113)
PVT	0.132	1.581 (0.871-2.871)
LN metastasis	0.867	1.092 (0.389-3.061)
Multiplicity	0.957	1.016 (0.561-1.841)
AFP	0.020	1.001 (1.000-1.002)	0.020	1.001 (1.000-1.002)
PIVKA-II	0.211	1.022 (0.988-1.056)
Pre-RT treatment	0.693	0.887 (0.487-1.608)
Baseline IL-1	0.262	0.979 (0.942-1.016)
Baseline IL-6	0.692	1.001 (0.996-1.007)
Baseline IL-8	0.770	1.000 (0.998-1.002)
Baseline IL-10	0.909	0.994 (0.890-1.109)
Baseline IL-12	0.449	1.014 (0.978-1.052)
Baseline TNF-α	0.573	0.984 (0.930-1.041)

Age, size, AFP, PIVKA-II, and baseline cytokine levels were analyzed as continuous variables using the Cox hazard ratio model. BCLC: Barcelona Clinic Liver Cancer; PVT: Portal vein thrombosis; LN: Lymph node; AFP: α-fetoprotein; PIVKA-II: Prothrombin induced by vitamin K absence-II; RT: Radiotherapy; IL: Interleukin; TNF: Tumor necrosis factor.

Table 3 Univariate and multivariate analyses of clinical factors or cytokines for failure-free survival - infield failure-free survival

	Univariate		Multivariate
	P value	RR (95%CI)	P value	RR (95%CI)
Sex	0.001	10.262 (2.479-42.481)	< 0.001	47.505 (7.384-305.601)
Age	0.198	0.968 (0.921-1.017)
Stage	0.082	2.106 (0.911-4.871)
BCLC	0.441	1.257 (0.702-2.250)
Size	0.903	1.007 (0.904-1.121)
PVT	0.247	1.741 (0.681-4.452)
LN metastasis	0.408	1.867 (0.425-8.191)
Multiplicity	0.944	1.033 (0.412-2.591)
AFP	0.653	0.999 (0.996-1.002)
PIVKA-II	0.889	0.996 (0.947-1.048)
Pre-RT treatment	0.290	1.627 (0.660-4.013)
Baseline IL-1	0.895	0.997 (0.955-1.041)
Baseline IL-6	0.000	1.011 (1.005-1.018)	< 0.001	1.019 (1.011-1.028)
Baseline IL-8	0.985	1.000 (0.997-1.003)
Baseline IL-10	0.437	1.060 (0.915-1.227)
Baseline IL-12	0.903	0.996 (0.938-1.059)
Baseline TNF-α	0.864	0.993 (0.918-1.074)

Age, size, AFP, PIVKA-II, and baseline cytokine levels were analyzed as continuous variables using the Cox hazard ratio model. BCLC: Barcelona Clinic Liver Cancer; PVT: Portal vein thrombosis; LN: Lymph node; AFP: α-fetoprotein; PIVKA-II: Prothrombin induced by vitamin K absence-II; RT: Radiotherapy; IL: Interleukin; TNF: Tumor necrosis factor.

Table 4 Univariate and multivariate analyses of clinical factors or cytokines for failure-free survival - outfield-intrahepatic failure-free survival

	Univariate		Multivariate
	P value	RR (95%CI)	P value	RR (95%CI)
Sex	0.387	1.539 (0.579-4.086)
Age	0.834	0.996 (0.959-1.035)
Stage	0.601	0.834 (0.423-1.646)
BCLC	0.383	1.210 (0.788-1.859)
Size	0.534	0.974 (0.898-1.058)
PVT	0.326	1.433 (0.699-2.940)
LN metastasis	0.387	0.530 (0.126-2.230)
Multiplicity	0.303	1.464 (0.709-3.021)
AFP	0.649	1.000 (0.999-1.002)
PIVKA-II	0.920	1.002 (0.963-1.043)
Pre-RT treatment	0.745	1.126 (0.550-2.307)
Baseline IL-1	0.573	1.015 (0.964-1.069)
Baseline IL-6	0.271	0.950 (0.985-1.004)
Baseline IL-8	0.942	1.000 (0.998-1.002)
Baseline IL-10	0.026	0.830 (0.705-0.978)	0.026	0.830 (0.705-0.978)
Baseline IL-12	0.197	0.959 (0.901-1.022)
Baseline TNF-α	0.240	0.951 (0.876-1.034)

Age, size, AFP, PIVKA-II, and baseline cytokine levels were analyzed as continuous variables using the Cox hazard ratio model. BCLC: Barcelona Clinic Liver Cancer; PVT: Portal vein thrombosis; LN: Lymph node; AFP: α-fetoprotein; PIVKA-II: Prothrombin induced by vitamin K absence-II; RT: Radiotherapy; IL: Interleukin; TNF: Tumor necrosis factor.

Table 5 Univariate and multivariate analyses of clinical factors or cytokines for failure-free survival - extrahepatic failure-free survival

	Univariate		Multivariate
	P value	RR (95%CI)	P value	RR (95%CI)
Sex	0.153	2.043 (0.768-5.438)
Age	0.502	0.987 (0.951-1.025)
Stage	0.196	1.562 (0.795-3.070)
BCLC	0.782	0.942 (0.615-1.441)
Size	0.830	1.010 (0.926-1.101)
PVT	0.988	0.994 (0.465-2.125)
LN metastasis	0.936	0.952 (0.289-3.141)
Multiplicity	0.622	1.197 (0.586-2.448)
AFP	0.034	1.001 (1.000-1.003)	0.034	1.001 (1.000-1.003)
PIVKA-II	0.449	1.016 (0.976-1.057)
Pre-RT treatment	0.878	1.057 (0.520-2.147)
Baseline IL-1	0.725	1.005 (0.977-1.034)
Baseline IL-6	0.529	1.002 (0.996-1.007)
Baseline IL-8	0.546	1.001 (0.999-1.003)
Baseline IL-10	0.309	1.062 (0.946-1.193)
Baseline IL-12	0.726	1.009 (0.962-1.058)
Baseline TNF-α	0.956	1.002 (0.942-1.066)

Age, size, AFP, PIVKA-II, and baseline cytokine levels were analyzed as continuous variables using the Cox hazard ratio model. BCLC: Barcelona Clinic Liver Cancer; PVT: Portal vein thrombosis; LN: Lymph node; AFP: α-fetoprotein; PIVKA-II: Prothrombin induced by vitamin K absence-II; RT: Radiotherapy; IL: Interleukin; TNF: Tumor necrosis factor.

Table 6 Variation of serum cytokine levels before and after radiotherapy (mean ± SEM)

	Before RT (pg/mL)	After RT (pg/mL)	P value
IL-1	6.77 ± 1.18	9.13 ± 1.51	0.229
IL-6	31.63 ± 7.34	29.54 ± 6.60	0.794
IL-8	115.75 ± 19.84	159.73 ± 38.96	0.211
IL-10	4.19 ± 0.41	5.83 ± 0.46	0.002
IL-12	6.10 ± 1.01	4.10 ± 0.59	0.018
TNF-α	4.27 ± 0.79	3.33 ± 0.40	0.130

IL: Interleukin; TNF: Tumor necrosis factor; RT: Radiotherapy.