High Inflammatory Factor Grading Predicts Poor Disease-Free Survival in AJCC Stage I-II Hepatocellular Carcinoma Patients After R0 Resection

Figures & data

Table 1 The Basic Characteristics for the Study Participants

Characteristics	All (n=87)
Gender, male/female	71/16
Age (y)	55.2±9.7
AFP (ng/L)	275.6±450.6
ALB (g/L)	38.6±4.0
ALT (U/L)	50.6±37.3
AST (U/L)	40.6±32.6
GGT (U/L)	70.1±96.3
PLT (10^9/L)	139.9±59.1
TBIL (μmol/L)	16.3±6.3
Tumor number, single/multiple	83/4
Tumor size (cm)	3.8±1.4
Tumor margin (mm)	9.4±9.6
Differentiation, high and middle/low	66/21
Microvascular invasion, yes/no	15/72
Capsule invasion, yes/no	56/31
HBV infection, yes/no	78/9
Liver cirrhosis, yes/no	10/77
Child-Pugh grade, A/B	84/3

Abbreviations: AFP, alpha-fetoprotein; ALB, albumin; ALT, alanine aminotransferase; AST, aspartate aminotransferase; GGT, gamma-glutamyl transpeptidase; PLT, platelet; TBIL, total bilirubin; HBV, hepatitis B virus.

Figure 1 Immunohistochemical analysis of IL-6, IL-6R, IL-1β, IKKβ, and P38MAPK protein levels in HCC tissue specimens (A) IL-6 expression (×400), (B) IL-6R expression (×400), (C) IL-1β expression (×400), (D) IKKβ expression (×400), (E) P38MAPK expression (×400).

Abbreviations: IL-6, interleukin-6; IL-6R, interleukin-6 receptor; IL-1β, interleukin-1 beta; IKKβ, inhibitor kappa B kinase beta; P38MAPK, p38 mitogen-activated protein kinase; HCC, hepatocellular carcinoma.

Figure 2 Kaplan-Meier survival plots comparing DFS for HCC patients stratified by low and high expression levels. (A) The DFS of patients with high IL-6 expression levels was worse than that of patients with low IL-6 expression levels (P=0.0065). (B) The DFS of patients with high IL-6R expression levels was worse than that of patients with low IL-6R expression levels (P=0.029). (C) The DFS of patients with high IL-1β expression levels was better than that of patients with low IL-1β expression levels (P=0.014). (D) The DFS of patients with high IKKβ expression levels was better than that of patients with low IKKβ expression levels (P=0.038). (E) The DFS of patients with high P38MAPK expression levels was better than that of patients with low P38MAPK expression levels (P=0.0033).

Abbreviations: DFS, disease-free survival; HCC, hepatocellular carcinoma; IL-6, interleukin-6; IL-6R, interleukin-6 receptor; IL-1β, interleukin-1 beta; IKKβ, inhibitor kappa B kinase beta; P38MAPK, p38 mitogen-activated protein kinase.

Table 2 The Median DFS Rates of the Low and High Expression Groups of Different Inflammatory Factors

Characteristics	Median DFS (95% CI, Months)		Log Rank test	P
Low	High
IL-6	35.0 (0.0–90.4)	10.2 (7.4–13.0)	7.396	0.007^a
IL-6R	23.3 (9.0–37.6)	10.2 (6.0–14.5)	4.790	0.029^a
JAK2	11.6 (5.7–17.5)	29.5 (0.0–64.8)	0.994	0.319
STAT3	12.9 (5.2–20.7)	10.8 (0.0–32.4)	1.055	0.304
SOCS3	11.7 (1.8–21.6)	12.9 (0.0–30.3)	0.284	0.594
IL-1α	12.9 (0.0–29.0)	11.9 (3.3–20.5)	0.045	0.832
IL-1β	9.3 (7.0–11.6)	23.3 (9.7–36.9)	6.099	0.014^a
IL-8	21.5 (7.8–35.2)	10.5 (8.1–12.9)	0.461	0.497
TNF-α	9.3 (6.8–11.8)	23.3 (6.9–39.7)	2.113	0.146
NFκB	9.3 (5.1–13.5)	18.1 (0.0–37.5)	1.877	0.171
IKKβ	10.2 (6.2–14.2)	23.3 (0.0–49.7)	4.289	0.038^a
P38MAPK	7.1 (6.5–7.7)	23.3 (16.6–30.0)	8.639	0.003^a

Note: ^aStatistically significant.

Abbreviations: DFS, disease-free survival; CI, confidence interval; IL-6, interleukin-6; IL-6R, interleukin-6 receptor; JAK2, janus kinase 2; STAT3, signal transducer and activator of transcription 3; SOCS3, Suppressor of cytokine signaling 3; IL-1α, interleukin-1 alpha; IL-1β, interleukin-1 beta; IL-8, interleukin-8; TNF-α, tumor necrosis factor-alpha; NFκB, nuclear factor kappa-light-chain-enhancer of activated B cells; IKKβ, inhibitor kappa B kinase beta; P38MAPK, p38 mitogen-activated protein kinase.

Table 3 The Assignment of IHC Scores and Establishment of IFG

Characteristics	Assignment		Score	IFG
Low	High
IL-6	1	2	5,6,7	A
IL-6R	1	2	8,9,10	B
IL-1β	2	1
IKKβ	2	1
P38MAPK	2	1

Abbreviations: IHC, immunohistochemistry; IFG, inflammatory factor grade; IL-6, interleukin-6; IL-6R, interleukin-6 receptor; IL-1β, interleukin-1 beta; IKKβ, inhibitor kappa B kinase beta; P38MAPK, p38 mitogen-activated protein kinase.

Table 4 Basic Characteristics of the Study Participants in the IFG-A and IFG-B Groups

Characteristics	IFG-A (n=59)	IFG-B (n=28)	T/χ^2 Test	P
Gender, male/female	45/14	26/2	3.480	0.062
Age(y)	55.5±9.4	54.8±10.4	0.318	0.752
AFP (ng/L)	297.7±463.1	228.9±427.5	0.663	0.509
ALB (g/L)	38.8±4.2	38.1±3.6	0.767	0.445
ALT (U/L)	46.8±35.9	58.5±39.7	−1.323	0.192
AST (U/L)	37.5±30.8	47.3±35.9	−1.316	0.192
GGT (U/L)	69.8±105.4	70.9±75.1	−0.052	0.958
PLT (10^9/L)	137.4±62.3	145.3±52.3	−0.585	0.560
TBIL (μmol/L)	17.0±6.5	14.8±5.6	1.525	0.131
Tumor number, single/multiple	58/1	25/3	1.766	0.184
Tumor size(cm)	3.5±1.1	4.3±1.9	−2.168	0.037^a
Tumor margin(mm)	10.4±8.9	7.4±10.7	1.382	0.171
Differentiation, high and middle/low	48/11	18/10	3.022	0.082
Microvascular invasion, yes/no	10/49	5/23	0.000	1.000
Capsule invasion, yes/no	36/23	20/8	0.897	0.343
HBV infection, yes/no	53/6	25/3	0.000	1.000
Liver cirrhosis, yes/no	9/50	1/27	1.529	0.216
Child-Pugh grade, A/B	57/2	27/1	-	1.000

Note: ^aStatistically significant.

Abbreviations: IFG, inflammatory factor grade; AFP, alpha-fetoprotein; ALB, albumin; ALT, alanine aminotransferase; AST, aspartate aminotransferase; GGT, gamma-glutamyl transpeptidase; PLT, platelet; TBIL, total bilirubin; HBV, hepatitis B virus.

Figure 3 Kaplan-Meier survival plots comparing DFS and OS for HCC patients stratified into IFG-A and IFG-B groups. (A) The DFS of patients in the IFG-B group was worse than that of patients in the IFG-A group (P<0.001). (B) The OS of patients in the IFG-B group was not significantly different from that in the IFG-A group (P=0.2).

Abbreviations: DFS, disease-free survival; OS, overall survival; IFG, inflammatory factor grade.

Table 5 Univariate and Multivariate Analyses of Disease-Free Survival

Variable	Univariate		Multivariate
HR (95% CI)	P value	HR (95% CI)	P value
Gender, male/female	0.647 (0.329–1.272)	0.206
Age (y, <50 vs ≥50)	1.138 (0.659–1.963)	0.643
AFP (ng/L, ≤400 vs.>400)	1.276 (0.724 −2.249)	0.399
ALB (g/L, ≤35 vs.>35)	0.748 (0.341–1.642)	0.470
ALT (U/L, ≤60 vs.>60)	1.763 (0.971–3.201)	0.062
AST (U/L, ≤42 vs.>42)	1.968 (1.152–3.362)	0.013^a
GGT (U/L, ≤60 vs.>60)	1.199 (0.710–2.023)	0.497
PLT (10^9/L, ≤100 vs.>100)	0.552 (0.319–0.955)	0.034^a	0.522 (0.299–0.909)	0.022^a
TBIL (μmol/L, ≤22 vs.>22)	1.742 (0.885–3.428)	0.108
Tumor number, single/multiple	2.786 (1.003–7.738)	0.049^a
Tumor size (cm, <5 vs ≥5)	2.278 (1.297–4.002)	0.004^a	2.362 (1.317–4.236)	0.004^a
Tumor margin (cm, <2 vs ≥2)	0.464 (0.185–1.159)	0.100
Differentiation, high and middle/low	1.951 (1.117–3.410)	0.019^a	1.966 (1.105–3.498)	0.022^a
Microvascular invasion, yes/no	1.732 (0.941–3.190)	0.078
Capsule invasion, yes/no	1.137 (0.678–1.907)	0.625
HBV infection	1.101 (0.474–2.557)	0.823
Liver cirrhosis, yes/no	1.134 (0.540–2.381)	0.739
Child-Pugh grade, A/B	1.363 (0.333–5.586)	0.667
IFG, A/B	2.677 (1.586–4.521)	<0.001^a	2.535 (1.468–4.378)	0.001^a

Note: ^aStatistically significant.

Abbreviations: HR, hazard ratio; CI, confidence interval; AFP, alpha-fetoprotein; ALB, albumin; ALT, alanine aminotransferase; AST, aspartate aminotransferase; GGT, gamma-glutamyl transpeptidase; PLT, platelet; TBIL, total bilirubin; HBV, hepatitis B virus; IFG, inflammatory factor grade.

Table 6 Univariate and Multivariate Analyses of Overall Survival

Variable	Univariate		Multivariate
HR (95% CI)	P value	HR (95% CI)	P value
Gender, male/female	0.718 (0.303–1.701)	0.451
Age (y, <50 vs ≥50)	1.106 (0.575–2.130)	0.762
AFP (ng/L, ≤400 vs.>400)	1.030 (0.505–2.098)	0.936
ALB (g/L, ≤35 vs.>35)	0.627 (0.223–1.763)	0.376
ALT (U/L, ≤60 vs.>60)	1.318 (0.649–2.678)	0.445
AST (U/L, ≤42 vs.>42)	2.032 (1.092–3.782)	0.025^a	1.933 (1.031–3.623)	0.040^a
GGT (U/L, ≤60 vs.>60)	1.596 (0.864–2.946)	0.135
PLT (10^9/L, ≤100 vs.>100)	0.484 (0.254–0.920)	0.027^a
TBIL (μmol/L, ≤22 vs.>22)	1.562 (0.690–3.538)	0.285
Tumor number, single/multiple	2.070 (0.636–6.741)	0.227
Tumor size (cm, ≤5 vs.>5)	1.640 (0.824–3.264)	0.159
Tumor margin (cm, <2 vs ≥2)	0.293 (0.071–1.213)	0.090
Differentiation, high and middle/low	2.448 (1.298–4.616)	0.006^a	2.402 (1.268–4.500)	0.007^a
Microvascular invasion, yes/no	1.770 (0.846–3.703)	0.129
Capsule invasion, yes/no	0.830 (0.450–1.532)	0.551
HBV infection	2.302 (0.964–5.494)	0.060
Liver cirrhosis, yes/no	1.773 (0.744–4.225)	0.196
Child-Pugh grade, A/B	1.394 (0.191–10.148)	0.743
IFG, A/B	1.489 (0.810–2.736)	0.200

Note: ^aStatistically significant.

Abbreviations: HR, hazard ratio; CI, confidence interval; AFP, alpha-fetoprotein; ALB, albumin; ALT, alanine aminotransferase; AST, aspartate aminotransferase; GGT, gamma-glutamyl transpeptidase; PLT, platelet; TBIL, total bilirubin; HBV, hepatitis B virus; IFG, inflammatory factor grade.