High Endothelin Receptor Type A Expression as an Independent Prognostic Biomarker and Correlated with Immune Infiltrates in Stomach Adenocarcinoma

Figures & data

Figure 1 High EDNRA expression is correlated with clinicopathologic features in patients with STAD. (A) Wilcoxon signed rank sum test was used to analyze the difference expression of EDNRA in STAD tissues and adjacent STAD tissues; (B) Wilcoxon signed rank sum test was used to detect the different expression of EDNRA in normal samples of GTEx combined adjacent STAD tissues and STAD samples; (C) the different expression of EDNRA in 27 STAD samples and matched adjacent samples; (D) ROC curve for EDNRA in normal samples of GTEx combined adjacent STAD tissues and STAD samples.

Table 1 Correlation Between EDNRA Expression and Clinicopathologic Characteristics of Patients with STAD

Characteristic	Low Expression of EDNRA N(%)	High Expression of EDNRA N(%)
n	187	188
Age, n (%)
≤65	82 (22.1%)	82 (22.1%)
>65	103 (27.8%)	104 (28%)
Gender, n (%)
Female	71 (18.9%)	63 (16.8%)
Male	116 (30.9%)	125 (33.3%)
Pathologic stage, n (%)
Stage I	33 (9.4%)	20 (5.7%)
Stage II	54 (15.3%)	57 (16.2%)
Stage III	72 (20.5%)	78 (22.2%)
Stage IV	18 (5.1%)	20 (5.7%)
T stage, n (%)
T1	14 (3.8%)	5 (1.4%)
T2	50 (13.6%)	30 (8.2%)
T3	84 (22.9%)	84 (22.9%)
T4	39 (10.6%)	61 (16.6%)
M stage, n (%)
M0	165 (46.5%)	165 (46.5%)
M1	13 (3.7%)	12 (3.4%)
N stage, n (%)
N0	54 (15.1%)	57 (16%)
N1	56 (15.7%)	41 (11.5%)
N2	39 (10.9%)	36 (10.1%)
N3	32 (9%)	42 (11.8%)
Histological type, n (%)
Diffuse Type	30 (8%)	33 (8.8%)
Mucinous Type	5 (1.3%)	14 (3.7%)
Not Otherwise Specified	98 (26.2%)	109 (29.1%)
Papillary Type	2 (0.5%)	3 (0.8%)
Signet Ring Type	5 (1.3%)	6 (1.6%)
Tubular Type	47 (12.6%)	22 (5.9%)
Residual tumor, n (%)
R0	153 (46.5%)	145 (44.1%)
R1	6 (1.8%)	9 (2.7%)
R2	9 (2.7%)	7 (2.1%)
Histologic grade, n (%)
G1	6 (1.6%)	4 (1.1%)
G2	81 (22.1%)	56 (15.3%)
G3	97 (26.5%)	122 (33.3%)
Reflux history, n (%)
No	98 (45.8%)	77 (36%)
Yes	23 (10.7%)	16 (7.5%)
Antireflux treatment, n (%)
No	73 (40.8%)	69 (38.5%)
Yes	17 (9.5%)	20 (11.2%)
H. pylori infection, n (%)
No	90 (55.2%)	55 (33.7%)
Yes	7 (4.3%)	11 (6.7%)
Barrett’s esophagus, n (%)
No	115 (55.3%)	78 (37.5%)
Yes	11 (5.3%)	4 (1.9%)
Primary therapy outcome, n (%)
PD	40 (12.6%)	25 (7.9%)
SD	8 (2.5%)	9 (2.8%)
PR	2 (0.6%)	2 (0.6%)
CR	114 (36%)	117 (36.9%)

Abbreviations: CR, complete response; PD, progressive disease; SD, stable disease; PR, partial response.

Table 2 EDNRA Expression Associated with Clinicopathologic Characteristics (Logistic Regression)

Characteristics	Total (N)	Odds Ratio (OR)	P value
Age (>65 vs ≤65)	371	1.313 (0.871–1.983)	0.194
Gender (Male vs Female)	375	2.042 (1.331–3.155)	0.001
Pathologic stage (Stage III&Stage IV vs Stage I&Stage II)	352	0.997 (0.655–1.516)	0.988
T stage (T3&T4 vs T2&T1)	367	0.568 (0.354–0.905)	0.018
M stage (M1 vs M0)	355	0.946 (0.414–2.145)	0.893
N stage (N1 vs N0)	208	0.895 (0.518–1.544)	0.690
Residual tumor (R1&R2 vs R0)	329	1.312 (0.624–2.828)	0.477
Histologic grade (G2&G3 vs G1)	366	1.551 (0.436–6.156)	0.502
Reflux history (Yes vs No)	214	0.673 (0.333–1.351)	0.266
H. pylori infection (Yes vs No)	163	0.235 (0.072–0.660)	0.009
Antireflux treatment (Yes vs No)	179	0.693 (0.333–1.435)	0.322
Barrett’s esophagus (Yes vs No)	208	0.827 (0.286–2.445)	0.723
Primary therapy outcome (PR&CR vs PD&SD)	317	0.563 (0.336–0.935)	0.027

Abbreviations: CR, complete response; PD, progressive disease; SD, stable disease; PR, partial response.

Figure 2 Box plot assessing EDNRA expression of patients with STAD according to different clinical characteristics. (A) T classification; (B) H. pylori infection; (C) pathologic stage.

Table 3 Associations with Clinicopathologic Characteristics in STAD Patients Using Cox Regression

Characteristics	Total(N)	Univariate Analysis		Multivariate Analysis
		Hazard Ratio (95% CI)	P value	Hazard Ratio (95% CI)	P value
Age (>65 vs ≤65)	367	1.620 (1.154–2.276)	0.005	1.681 (1.096–2.577)	0.017
Gender (Male vs Female)	370	1.267 (0.891–1.804)	0.188
T stage (T3&T4 vs T1&T2)	362	1.719 (1.131–2.612)	0.011	1.144 (0.561–2.334)	0.712
M stage (M1 vs M0)	352	2.254 (1.295–3.924)	0.004	1.482 (0.662–3.321)	0.339
N stage (N1&N2&N3 vs N0)	352	1.925 (1.264–2.931)	0.002	1.948 (1.012–3.749)	0.046
Pathologic stage (Stage II&Stage III&Stage IV vs Stage I)	347	2.247 (1.210–4.175)	0.010	1.238 (0.366–4.182)	0.731
Residual tumor (R1&R2 vs R0)	325	3.445 (2.160–5.494)	<0.001	1.433 (0.780–2.631)	0.247
Reflux history (Yes vs No)	213	0.582 (0.291–1.162)	0.125
H. pylori infection (Yes vs No)	162	0.650 (0.279–1.513)	0.317
Barrett’s esophagus (Yes vs No)	207	0.892 (0.326–2.441)	0.824
Histologic grade (G2&G3 vs G1)	361	1.957 (0.484–7.910)	0.346
Antireflux treatment (Yes vs No)	179	0.756 (0.422–1.353)	0.346
Primary therapy outcome (PR&CR vs PD&SD)	313	0.244 (0.168–0.354)	<0.001	0.207 (0.132–0.324)	<0.001
EDNRA (High vs Low)	370	1.537 (1.105–2.139)	0.011	2.414 (1.568–3.715)	<0.001

Abbreviations: CR, complete response; PD, progressive disease; SD, stable disease; PR, partial response.

Figure 3 The independent risk and diagnostic value of EDNRA expression in STAD. (A) The survival analysis of overall survival (OS) of TCGA-STAD database; (B) Kaplan–Meier survival analysis of OS; (C) free progression (FP); (D) post progression survival (PPS).

Figure 4 A nomogram for predicting probability of patients with 1-, 3- and 5-year overall survival (OS).

Figure 5 Forest plot of the multivariate Cox regression analysis in TCGA-STAD.

Figure 6 Enrichment plots from GSEA. (A–C) EDNRA-related signaling pathways in c2.cp.v7.2.symbols.gmt; (D–F) EDNRA-related signaling pathways in c5.all.v7.2.symbols.gmt (Gene ontology).

Figure 7 Immune Cell Infiltration Analysis of EDNRA in the TCGA-STAD. (A) The forest plot shows the correlation between EDNRA expression level and 24 immune cells; (B) the enrichment scores of EDNRA expression in NK cells and macrophages; (C) The correlation between EDNRA expression and macrophages; (D) the correlation between EDNRA expression and NK cells.

Abbreviations: Tcm, Tcentral memory; Tem, Teffector memory; Tgd, Tgamma delta; Tfh, Tfollicular helper; NK, natural killer; pDC, plasmacytoid dendritic cells.

Figure 8 EDNRA expression levels in different types of cancers. (A) Compared with adjacent tissues, increased or decreased expression of EDNRA in Oncomine. (B) EDNRA expression levels in different types of cancers from TCGA datasets in TIMER, **P <0.01, ***P < 0.001.

Figure 9 qRT-PCR of EDNRA expression in STAD cell lines (MGC-803, MKN-45) and human normal gastric epithelial cell line GES-1; *P < 0.05, ***P < 0.001.