Diagnostic and prognostic roles of IRAK1 in hepatocellular carcinoma tissues: an analysis of immunohistochemistry and RNA-sequencing data from the cancer genome atlas

Figures & data

Figure 1 The IHC of normal liver tissues.

Notes: IRAK1 negative staining in normal liver tissues (A: ×100 and B: ×400). IRAK1 positive staining in normal liver tissues (C: ×100 and D: ×400).
Abbreviation: IHC, immunohistochemistry.

Figure 2 The IHC of liver tissues with cirrhosis.

Notes: IRAK1 negative staining in cirrhosis tissues (A: ×100 and B: ×400). IRAK1 positive staining in cirrhosis tissues (C: ×100 and D: ×400).
Abbreviation: IHC, immunohistochemistry.

Figure 3 The IHC of para-HCC tissues with cirrhosis.

Notes: IRAK1 negative staining in cirrhosis tissues (para-HCC; A: ×100 and B: ×400). IRAK1 positive staining in cirrhosis tissues (para-HCC; C: ×100 and D: ×400).
Abbreviations: IHC, immunohistochemistry; HCC, hepatocellular carcinoma.

Figure 4 The IHC of normal liver tissues (para-HCC).

Notes: IRAK1 negative staining in normal liver tissues (para-HCC; A: ×100 and B: ×400). IRAK1 positive staining in normal liver tissues (para-HCC; C: ×100 and D: ×400).
Abbreviations: IHC, immunohistochemistry; HCC, hepatocellular carcinoma.

Figure 5 The IHC of HCC tissues.

Notes: IRAK1 negative staining in HCC tissues (A: ×100 and B: ×400). IRAK1 positive staining in HCC tissues (C: ×100 and D: ×400).
Abbreviations: IHC, immunohistochemistry; HCC, hepatocellular carcinoma.

Figure 6 The expression of IRAK1 in IHC.

Notes: (A) IRAK1 expression in different types of liver tissues. The highest IRAK1 expression was observed in HCC (48.5%) compared with para-HCC tissues (34.5%, P=0.008), liver cirrhosis tissues (24.3%, P=0.007) and normal liver tissues (15.2%, P<0.001). Moreover, the IRAK1 expression increased with the deterioration from noncancerous liver disease to HCC (P<0.001). (B) Tumor size and (C) tumor metastasis. IRAK1 expression was found to be higher in the group of patients with a large tumor size (54.0%) and metastasis (56.8%) than in the group of patients with a small tumor size (37.9%, P=0.047) and no metastasis (41.1%, P=0.041).
Abbreviations: IHC, immunohistochemistry; HCC, hepatocellular carcinoma.

Table 1 Expression of IRAK1 in different types of liver tissues

Tissue	Patients (n)	IRAK1 negative, n (%)	IRAK1 positive, n (%)	χ^2	P-value
HCC vs	171	88 (51.5)	83 (48.5)
Para-carcinoma tissue	171	112 (65.5)	59 (34.5)	6.936	0.008
Liver cirrhosis	37	28 (75.7)	9 (24.3)	7.230	0.007
Normal	33	28 (84.8)	5 (15.2)	12.570	,0.001
Para-carcinoma tissue vs	171	112 (65.5)	59 (34.5)
Liver cirrhosis	37	28 (75.7)	9 (24.3)	1.432	0.231
Normal	33	28 (84.8)	5 (15.2)	4.811	0.028
Liver cirrhosis vs	37	28 (75.7)	9 (24.3)
Normal	33	28 (84.8)	5 (15.2)	0.917	0.338

Note: Linear by linear association test (χ²=18.871, P<0.001) among normal, liver cirrhosis and para-carcinoma tissues, and HCC revealed an increasing trend of IRAK1 expression in these four tissues.

Abbreviation: HCC, hepatocellular carcinoma.

Figure 7 The ROC curves of IRAK1 expression by IHC in HCC.

Notes: The ROC curves indicated that IRAK1 (blue line) showed a weak diagnostic ability for HCC (AUC =0.591, P=0.002). Green line denotes reference line.
Abbreviations: ROC, receiver operating characteristic; IHC, immunohistochemistry; HCC, hepatocellular carcinoma; AUC, area under the curve.

Table 2 Relationship between IRAK1 expression and clinicopathological features in HCC

Clinicopathological parameters	n	IRAK1 negative, n (%)	IRAK1 positive, n (%)	χ^2	P-value	Correlation
						r	P-value
Age (years)					0.431		0.432
<50	67	37	30	−0.788		0.060
$50	104	51	53
Gender					0.713		0.715
Male	153	78 (51.0)	75 (49.0)	0.135		−0.028
Female	18	10 (55.6)	8 (44.4)
Edmondson grade					0.199		0.173
High	20	14 (70.0)	6 (30.0)	3.226		0.105
Moderate	98	49 (50.0)	49 (50.0)
Low	53	25 (47.2)	28 (52.8)
Nodes					0.679		0.700
Single	68	30 (44.1)	38 (55.9)	0.152		−0.034
Multiple	61	29 (47.5)	32 (52.5)
Tumor size (cm)					0.047		0.047
<5	58	36 (62.1)	22 (37.9)	3.953		0.152
≥5	113	52 (46.0)	61 (54.0)
TNM stage					0.071		0.072
I–II	48	30 (62.5)	18 (37.5)	3.255		0.138
III–IV	123	58 (47.2)	65 (52.8)
Portal vein tumor thrombus					0.829		0.831
Negative	84	39 (46.4)	45 (53.6)	0.046		0.019
Positive	45	20 (44.4)	25 (55.6)
Vaso-invasion					0.938		0.938
No	77	35 (45.5)	42 (54.5)	0.006		−0.007
Yes	52	24 (46.2)	28 (53.8)
Capsular					0.972		0.972
Complete	61	28 (45.9)	33 (54.1)	0.001		0.003
No or infiltrate	68	31 (45.6)	37 (54.4)
Metastasis					0.041		0.041
No	90	53 (58.9)	37 (41.1)	4.196		0.157
Yes	81	35 (43.2)	46 (56.8)
AFP					0.453		0.457
Positive	54	27 (50.0)	27 (50.0)	0.564		0.072
Negative	56	24 (42.9)	32 (57.1)

Abbreviations: AFP, alpha fetoprotein; HCC, hepatocellular carcinoma; TNM, tumor–node–metastasis.

Figure 8 The expression of IRAK1in HCC in the TCGA data.

Notes: (A) The expression of IRAK1 was upregulated in 354 cases of HCC tissues (11.5503±0.7946) compared with 50 adjacent normal liver tissues (10.3058±0.5212, P<0.001). (B) The paired t-test on 49 pairs of HCC tissues and the corresponding adjacent normal liver tissues confirmed this finding (11.6174±0.8979 vs 10.2921±0.5174, P<0.001). (C) Moreover, this significant difference in IRAK1 expression was observed only in the advanced tumor stage (T2–T4: 11.6363±0.8153 vs T1: 11.4647±0.7677, P=0.043).
Abbreviations: HCC, hepatocellular carcinoma; TCGA, the cancer genome atlas.

Figure 9 The ROC curve and Kaplan–Meier survival curve of IRAK1 expression for HCC from TCGA data.

Notes: (A) ROC curve. An AUC value of 0.910 indicated a strong diagnostic value (95% CI: 0.878–0.942, P<0.001). (B) Kaplan–Meier survival curve. The median survival time was 45.7 months for the IRAK1 high-expression group and 71.0 months for the IRAK low-expression group (log rank: P=0.008).
Abbreviations: ROC, receiver operating characteristic; HCC, hepatocellular carcinoma; TCGA, the cancer genome atlas; AUC, area under the curve; Cum, cumulative.

Table 3 Expression of IRAK1 detected by RNA sequencing and clinicopathological parameters in HCC in TCGA

Clinicopathological parameters	n	IRAK1 expression
		Mean ± SD	t	P-value
Tissue (unmatched)				<0.001
Adjacent noncancerous liver	50	10.3058±0.5212	14.649
HCC	354	11.5503±0.7946
Tissue (matched)				<0.001
Adjacent noncancerous liver	49	10.2921±0.5174	10.703
HCC	49	11.6174±0.8979
Age (years)				0.497
<60	163	11.5168±0.7733	−0.680
≥60	190	11.5746±0.8132
Gender				0.116
Male	241	11.5958±0.8123	1.575
Female	113	11.4534±0.7499
AJCC pathologic T				0.043
T1	174	11.4647±0.7677	−2.029
T2–T4	177	11.6363±0.8153
AJCC pathologic N				0.225
N0	241	11.5107±0.7965	F=1.498*
N1	3	11.1300±0.4606
NX	109	11.6453±0.7940
AJCC pathologic M				0.149
M0	256	11.5036±0.7794	F=1.912*
M1	4	11.3702±0.6400
MX	94	11.6853±0.8317
Pathologic stage				0.135
I–II	245	11.4982±0.7800	−1.499
III–IV	83	11.6407±0.8398
Vaso-invasion				0.704
−	196	11.4945±0.7900	−0.380
+	105	11.5311±0.7961

Note:

* One-way ANOVA was performed.

Abbreviations: HCC, hepatocellular carcinoma; TCGA, the cancer genome atlas; AJCC, American Joint Committee on Cancer; ANOVA, analysis of variance; SD, standard deviation.

Figure 10 The alteration of IRAK1 and its interaction in altered neighboring genes in 440 HCC cases from the cBioPortal database.

Notes: (A) IRAK1 alteration. In this cohort, 29% (126/440) of HCC cases exhibited IRAK1 alteration, including 85 cases of mRNA upregulation, 35 cases of mRNA downregulation, eight cases of amplification, three cases of missense mutation, two cases of truncating mutation and one case of deep deletion. Among them, eight cases possessed a mixed type of alteration. (B) Network of IRAK1 and altered neighboring genes. The network reflected easily identified interactions between IRAK1 and several genes, including MAP3K7, MAP2K4, NFKB1, AKT1 and ERC1.
Abbreviation: HCC, hepatocellular carcinoma.

Figure 11 Correlation between IRAK1 and MAPKAPK2.

Note: Pearson correlation analysis revealed that IRAK1 expression was significantly correlated with MAPKAPK2 expression (r=0.34, P<0.05).
Abbreviation: RNA-seq, RNA sequencing.

Figure 12 Analysis of the overall survival (A) and disease-free survival (B) of HCC patients with and without IRAK1 alteration.

Note: No statistically significant difference was identified.
Abbreviation: HCC, hepatocellular carcinoma.