Overexpression of EWSR1 (Ewing sarcoma breakpoint region 1/EWS RNA binding protein 1) predicts poor survival in patients with hepatocellular carcinoma

Figures & data

Figure 1. EWSR1 expression between tumor and non-tumor liver tissues in HCC patients in TCGA (a) and GEO (b) datasets, and our cohort (c). Hematoxylin and eosin staining of HCC (left) and paratumoral, healthy (right) tissues (d). Positive immunohistochemical staining of EWSR1 in HCC (right), negative in paratumoral normal (left) tissue (e). Scale: 100 μm. Abbreviations: GEO, Gene Expression Omnibus; HCC, hepatocellular carcinoma; TCGA, The Cancer Genome Atlas

Table 1. Correlation between EWSR1 expression and clinicopathological parameters of HCC patients in the TCGA cohort

Characteristics	EWSR1 expression		P
	High (n = 185)	Low (n = 184)
Histologic grade, n (%)
G1	17 (9.2)	38 (20.7)	< 0.001
G2	82 (44.3)	95 (51.6)
G3	77 (41.7)	44 (23.9)
G4	6 (3.2)	5 (2.7)
Not reported	3(1.6)	2(1.1)
Pathologic_M, n (%)
M0	133 (71.9)	131 (71.2)	0.989
M1	2 (1.1)	2 (1.1)
MX	50 (27.0)	51 (27.7)
Pathologic_N, n (%)
N0	128 (69.2)	122 (66.3)	0.426
N1	3 (1.6)	1 (0.5)
NX	54 (29.2)	61 (33.2)
Pathologic_T, (%)
T1	78 (42.2)	101 (54.9)	0.026
T2	50 (27.0)	44 (23.9)
T3	49 (26.5)	31 (16.9)
T4	8 (4.3)	5 (2.7)
TX	0 (0.0)	3(1.6)
Days to death, mean (SD)	560.73 (609.94)	813.12 (753.01)	0.039
Sex, n (%)
Female	65 (35.1)	55 (29.9)	0.335
Male	120 (64.9)	129 (70.1)
Race, n (%)
American Indian or Alaska native	1 (0.5)	1 (0.5)	0.377
Asian	83 (44.9)	74 (40.2)
Black or African American	9 (4.9)	8 (4.4)
Not reported	2 (1.1)	8 (4.4)
White	90 (48.6)	93 (50.5)
Vita status, n (%)
Alive	107 (57.8)	133 (72.3)	0.007
Dead	78 (42.2)	50 (27.2)
Not Reported	0 (0.0)	1 (0.5)
Age, mean (SD)	58.94 (13.49)	60.57 (13.30)	0.246
Tumor stage, n (%)
Not reported	13 (7.0)	11 (6.0)	0.068
Stage I	73 (39.5)	96 (52.2)
Stage II	44 (23.8)	42 (22.8)
Stage III	53 (28.6)	32 (17.4)
Stage IV	2 (1.1)	3 (1.6)

Abbreviations: HCC, Hepatocellular carcinoma; M, metastasis; N, nodule; T, tumor; TCGA, The Cancer Genome Atlas.

Table 2. Correlation between EWSR1 expression and clinicopathological parameters of HCC patients in our cohort

Characteristics n (%)	EWSR1 expression		χ^2	P
	Low (n = 64)	High (n = 60)
Sex			0.042	0.838
Male	58 (90.6)	55 (91.7)
Female	6 (9.4)	5 (8.3)
Age, mean (SD)			1.054	0.305
≤ 50 years	15 (23.4)	19 (31.7)
> 50 years	49 (76.6)	41 (68.3)
Tumor size			2.939	0.230
≤ 2 cm	6 (9.4)	3 (5.0)
2–5 cm	28 (43.8)	20 (33.3)
> 5 cm	30 (46.9)	37 (61.7)
Capsule invasion			1.472	0.225
No	7 (10.9)	3 (5.0)
Yes	57 (89.1)	57 (95.0)
Lymphovascular infiltration			2.160	0.142
No	11 (17.2)	5 (8.3)
Yes	53 (82.8)	55 (91.7)
Differentiation degree			7.319	0.026
Poor	5 (7.8)	15 (25.0)
Moderate	56 (87.5)	44 (73.3)
Good	3 (4.7)	1 (1.7)
Cirrhosis			3.228	0.072
No	14 (21.9)	6 (10.0)
Yes	50 (78.1)	54 (90.0)
T stage			13.327	0.004
T1	3 (4.7)	2 (3.3)
T2	30 (46.9)	15 (25.0)
T3	31 (48.4)	35 (58.4)
T4	0 (0)	8 (13.3)
N stage			6.991	0.008
N0	57 (89.1)	42 (70.0)
N1	7 (10.9)	18 (30.0)
TNM stage			17.348	0.001
I	3 (4.7)	2 (3.3)
II	28 (43.7)	9 (15.0)
III	30 (46.9)	35 (58.4)
IV	3 (4.7)	14 (23.3)
AFP expression			42.701	< 0.001
Low	47 (73.4)	9 (15.0)
High	17 (26.6)	51 (85.0)
Ki67 expression			16.814	< 0.001
Low	37 (57.8)	13 (21.7)
High	27 (42.2)	47 (78.3)
Recurrence			0.176	0.675
No	16 (25.0)	17 (28.3)
Yes	48 (75.0)	43 (71.7)
Death			22.642	< 0.001
No	41 (64.1)	13 (21.7)
Yes	23 (35.9)	47 (78.3)

Abbreviations: AFP, alpha-fetoprotein; HCC, Hepatocellular carcinoma; N, nodule; T, tumor; TNM; tumor-nodule-metastasis.

Table 3. Univariate and multivariate Cox regression analyses of DFS in HCC patients

Parameter	Univariate analysis		Multivariate analysis
	HR (95% CI)	P	HR (95% CI)	P
Gender (vs. male)	1.257 (0.544–2.901)	0.592
Age	0.673 (0.430–1.053)	0.083
Tumor size	1.348 (0.972–1.869)	0.073
Capsule invasion	1.880 (0.809–4.366)	0.142
Lymphovascular infiltration	1.662 (0.852–3.240)	0.136
Differentiation degree	0.962 (0.576–1.605)	0.882
Cirrhosis	1.710 (0.927–3.153)	0.086
T stage	1.413 (1.016–1.966)	0.040	1.184 (0.682–2.055)	0.549
N stage	2.226 (1.303–3.800)	0.003	1.498 (0.664–3.379)	0.330
TNM stage	1.542 (1.142–2.081)	0.005	1.121 (0.660–1.904)	0.672
AFP	3.133 (1.962–5.002)	< 0.001	2.339 (1.330–4.111)	0.003
Ki67	1.417 (0.921–2.181)	0.113
EWSR1 overexpression	3.177 (1.947–5.185)	< 0.001	1.654 (1.098–3.011)	0.041

Abbreviations: AFP, alpha-fetoprotein; CI, coefficient interval; DFS, disease-free survival; HCC, Hepatocellular carcinoma; HR, hazard ratio; N, nodule; T, tumor; TNM; tumor-nodule-metastasis.

Table 4. Univariate and multivariate Cox regression analyses of OS in HCC patients

Parameter	Univariate analysis		Multivariate analysis
	HR (95% CI)	P	HR (95% CI)	P
Gender (vs. male)	3.513 (1.325–9.315)	0.012
Age	0.938 (0.566–1.556)	0.805
Tumor size	1.526 (1.017–2.290)	0.041	0.769 (0.391–1.513)	0.447
Capsule invasion	4.089 (0.995–16.808)	0.051
Lymphovascular Infiltration	2.306 (0.990–5.369)	0.053
Differentiation degree	0.454 (0.275–0.750)	0.002	0.987 (0.525–1.856)	0.967
Cirrhosis	1.748 (0.888–3.439)	0.106
T stage	2.276 (1.479–3.502)	< 0.001	2.887 (1.374–6.065)	0.005
N stage	4.043 (2.269–7.202)	< 0.001	3.626 (1.510–8.706)	0.004
TNM stage	2.970 (1.972–4.473)	< 0.001	1.145 (0.644–2.036)	0.645
AFP	5.825 (3.184–10.654)	< 0.001	2.715 (1.337–5.516)	0.006
Ki67	1.753 (1.055–2.911)	0.030	0.932 (0.526–1.652)	0.811
EWSR1 overexpression	5.914 (3.472–10.074)	< 0.001	3.492 (1.824–6.685)	< 0.001

Abbreviations: AFP, alpha-fetoprotein; CI, coefficient interval; HCC, Hepatocellular carcinoma; HR, hazard ratio; N, nodule; OS, overall survival; T, tumor; TNM; tumor-nodule-metastasis.

Figure 2. Comparison of the 5- and 10-year overall survival between EWSR1 high and low groups in The Cancer Genome Atlas (TCGA) dataset (a, b) and our cohort (c, d)

Figure 3. Protein-protein interaction network of EWSR1 (a). Gene ontology (GO) (b) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway (c) analyses of coexpressed genes of EWSR1.