Kaplan-meier survival data of GC patients with differential expressions of Notch ligand can be discovered at www.kmplot.com. We were the first to analyze the prognostic value of DLL1 expression. The Affymetrix ID is: 224215_s_at. Figure 1A shows the survival curve of GC patients based on DLL1 mRNA expression level(n = 631). The ten-year survival rate of all GC patients with high expression level of DLL1 mRNA was worse, HR 1.27 (1.02–1.57), p = 0.032. The Affymetrix ID is: 227938_s_at. OS curves were plotted for carcinoma of stomach patients(n = 631) (Fig. 1B). The ten-year survival rate of all GC with high expression level of DLL1 was also worse, HR 1.75 (1.35–2.28), p = 2.1e-05.
For DLL3, the Affymetrix ID is: 219537_x_at. The OS of GC patients with high expression level of DLL3 mRNA was worse, HR 1.46 (1.23–1.73), p = 1.3e-05 (Fig. 2A), the same as the Affymetrix ID is: 222898_s_at, HR 1.49 (1.19–1.87), p = 0.00051 (Fig. 2B).
For DLL4, the Affymetrix ID is: 223525_at. High expression of DLL4 was significantly tied to worsening OS in GC patients, HR 1.35 (1.08–1.68), p = 0.0091 (Fig. 3).
For JAG1, the Affymetrix ID is: 216268_s_at. High expression of JAG1 was insignificant correlation with worsening OS in GC patients, HR 1.2 (0.99–1.46), p = 0.06 (Fig. 4A). Also, the same as the Affymetrix ID: 209098_s_at(Fig. 4B), the Affymetrix ID is: 209099_x_at(Fig. 4C) and the Affymetrix ID is: 216268_s_at(Fig. 4D).
For JAG2, the Affymetrix ID is: 209784_s_at. The OS of GC patients with high expression level of JAG2 mRNA was worse, HR 2.08 (1.69–2.55), p = 1.6e-12 (Fig. 5A). The same as Affymetrix ID is: 32137_at. HR 1.89 (1.57–2.29), p = 1.6e-11 (Fig. 5B).
Table-1 Correlation between expression of Notch ligands mRNA and gender in patients with GC
Notch ligands
|
Gender
|
Cases
|
HR 95% CI
|
P value
|
DLL1
|
Male
|
567
|
1.26 (0.94–1.70)
|
0.123
|
Female
|
244
|
0.64 (0.38–1.07)
|
0.083
|
DLL3
|
Male
|
567
|
1.71 (1.38–2.14)
|
1.2e-6*
|
Female
|
244
|
0.81 (0.55–1.20)
|
0.298
|
DLL4
|
Male
|
567
|
1.43(1.06–1.95)
|
0.020*
|
Female
|
244
|
1.29(0.84–2.00)
|
0.245
|
JAG1
|
Male
|
567
|
0.86 (0.69–1.07)
|
0.167
|
Female
|
244
|
0.75(0.53–1.06)
|
0.103
|
JAG2
|
Male
|
567
|
2.12(1.70–2.64)
|
7.3e-12*
|
Female
|
244
|
1.77 (1.24–2.52)
|
0.002*
|
Table-2 Correlation between expression of Notch ligands mRNA and clinical stages with GC
Notch ligands
|
clinical stages
|
Cases
|
HR 95% CI
|
P value
|
DLL1
|
Ⅰ
|
69
|
3.27 (0.87–11.7)
|
0.073
|
Ⅱ
|
145
|
0.51 (0.22–1.16)
|
0.101
|
Ⅲ
|
319
|
0.70 (0.48–1.02)
|
0.062
|
Ⅳ
|
152
|
0.82 (0.55–1.23)
|
0.338
|
DLL3
|
Ⅰ
|
69
|
0.29 (0.11–0.79)
|
0.010*
|
Ⅱ
|
145
|
2.37 (1.23–4.55)
|
0.008*
|
Ⅲ
|
319
|
1.56 (1.12–2.19)
|
0.009*
|
Ⅳ
|
152
|
0.75 (0.51–1.12)
|
0.157
|
DLL4
|
Ⅰ
|
69
|
0.65(0.22–1.94)
|
0.434
|
Ⅱ
|
145
|
1.45(0.77–2.72)
|
0.243
|
Ⅲ
|
319
|
1.4(0.91–2.17)
|
0.128
|
Ⅳ
|
152
|
1.49(1.01–2.22)
|
0.045*
|
JAG1
|
Ⅰ
|
69
|
0.33(0.12–0.91)
|
0.025*
|
Ⅱ
|
145
|
0.59(0.32–1.07)
|
0.081
|
Ⅲ
|
319
|
0.75(0.56–0.10)
|
0.052
|
Ⅳ
|
152
|
1.19(0.81–1.75)
|
0.370
|
JAG2
|
Ⅰ
|
69
|
3.58(1.32–9.69)
|
0.007*
|
Ⅱ
|
145
|
1.78(0.95–3.31)
|
0.066
|
Ⅲ
|
319
|
2.09(1.53–2.84)
|
1.8e-6*
|
Ⅳ
|
152
|
1.68(1.14–2.47)
|
0.009*
|
Table-3 Correlation between expression of Notch ligands mRNA and Lauren classification in patients with GC
Notch ligands
|
Lauren classification
|
Cases
|
HR 95% CI
|
P value
|
DLL1
|
intestinal
|
336
|
1.6 (1.02–2.57)
|
0.048*
|
diffuse
|
248
|
0.67 (0.45–1.00)
|
0.048
|
mixed
|
33
|
3.21 (0.71–14.54)
|
0.110
|
DLL3
|
intestinal
|
336
|
1.88 (1.34–2.64)
|
0.0002*
|
diffuse
|
248
|
0.72 (0.48–1.09)
|
0.125
|
mixed
|
33
|
0.43 (0.14–1.27)
|
0.114
|
DLL4
|
intestinal
|
336
|
0.81 (0.56–1.16)
|
0.253
|
diffuse
|
248
|
1.5 (1.06–2.13)
|
0.021*
|
mixed
|
33
|
4.18 (0.91–19.29)
|
0.047
|
JAG1
|
intestinal
|
336
|
0.63 (0.46–0.86)
|
0.004*
|
diffuse
|
248
|
1.38 (0.95–2.02)
|
0.089
|
mixed
|
33
|
0.16 (0.03–0.73)
|
0.008*
|
JAG2
|
intestinal
|
336
|
2.72 (1.97–3.74)
|
1.8e-10*
|
diffuse
|
248
|
1.66 (1.16–2.38)
|
0.005*
|
mixed
|
33
|
2.54 (0.89–7.26)
|
0.071
|
Table-4 Correlation between expression of Notch ligands mRNA and different differentiation in patients with GC
Notch ligands
|
differentiation
|
Cases
|
HR 95% CI
|
P value
|
DLL1
|
low differentiation adenocarcinoma
|
166
|
0.67 (0.38–1.17)
|
0.158
|
median differentiation adenocarcinoma
|
67
|
1.47 (0.71–3.04)
|
0.299
|
high differentiation adenocarcinoma
|
32
|
none
|
0.221
|
DLL3
|
low differentiation adenocarcinoma
|
166
|
0.61 (0.39–0.94)
|
0.024*
|
median differentiation adenocarcinoma
|
67
|
1.93 (0.98–3.77)
|
0.052
|
high differentiation adenocarcinoma
|
32
|
2.57 (1.03–6.39)
|
0.036*
|
DLL4
|
low differentiation adenocarcinoma
|
166
|
1.31 (0.78–2.19)
|
0.310
|
median differentiation adenocarcinoma
|
67
|
0.78 (0.41–1.5)
|
0.463
|
high differentiation adenocarcinoma
|
32
|
none
|
0.221
|
JAG1
|
low differentiation adenocarcinoma
|
166
|
0.59 (0.38–0.91)
|
0.017*
|
median differentiation adenocarcinoma
|
67
|
0.62 (0.28–1.35)
|
0.224
|
high differentiation adenocarcinoma
|
32
|
0.18 (0.04–0.79)
|
0.011*
|
JAG2
|
low differentiation adenocarcinoma
|
166
|
1.57 (0.95–2.6)
|
0.073
|
median differentiation adenocarcinoma
|
67
|
2.06 (0.98–4.32)
|
0.051
|
high differentiation adenocarcinoma
|
32
|
3.66 (1.32–10.15)
|
0.008*
|
Table-5 Correlation between expression of Notch ligands mRNA and different treatment of GC
Notch ligands
|
Treatment
|
Cases
|
HR 95% CI
|
P value
|
DLL1
|
Surgery
|
393
|
1.30 (0.91–1.85)
|
0.147
|
surgery combined with 5-FU chemotherapy
|
158
|
0.63 (0.23–1.69)
|
0.354
|
DLL3
|
Surgery
|
393
|
1.25 (0.93–1.66)
|
0.135
|
surgery combined with 5-FU chemotherapy
|
158
|
1.86 (1.30–2.67)
|
0.0006*
|
DLL4
|
Surgery
|
393
|
1.47 (1.06–2.04)
|
0.019*
|
surgery combined with 5-FU chemotherapy
|
158
|
2.92 (1.17–7.28)
|
0.016*
|
JAG1
|
Surgery
|
393
|
1.35 (1.01–1.8)
|
0.042*
|
surgery combined with 5-FU chemotherapy
|
158
|
0.77 (0.54–1.08)
|
0.128
|
JAG2
|
Surgery
|
393
|
1.54 (1.15–2.05)
|
0.003*
|
surgery combined with 5-FU chemotherapy
|
158
|
1.92 (1.34–2.76)
|
0.0003*
|
We filled the gaps in the association between individual Notch ligands and clinicopathological features, we separately associated gender (Table-1), clinical stages (Table-2), Lauren's classification (Table-3), different differentiation of GC patients (Table-4) and differences in treatment methods (Table-5). Just as Table 1, DLL3 high expression was so clearly tied to worsening OS in male, HR 1.71 (1.38–2.14), p = 1.2e-6. DLL4 mRNA expression was so clearly tied to worsening OS in male, HR 1.43(1.06–1.95), p = 0.02. JAG2 mRNA expression was so clearly tied to worsening OS in male, HR 2.12(1.70–2.64), p =7.3e-12 and female as well, HR 1.77 (1.24–2.52), p=0.002. According to Table 2, high expression of DLL3 was tied to worsen OS in advanced GC, such as stage Ⅱ, HR 2.37 (1.23–4.55), p=0.008 and stage Ⅲ, HR 1.56 (1.12–2.19), p =0.009. Nevertheless, DLL3 expression was linked with a better OS in grade Ⅰ, HR 0.29 (0.11–0.79), p = 0.01. DLL4 expression was only tied to worsening OS in stage Ⅳ, HR 1.49(1.01–2.22), p =0.045. JAG1 mRNA expression was so clearly tied to a better OS in stageⅠ, HR 0.33(0.12–0.91), p =0.025. JAG2 expression was so clearly tied to worsening OS in stageⅠ, HR 3.58(1.32–9.69), p= 0.007, stage Ⅲ, HR 2.09(1.53–2.84), p = 1.8e-6 and stage Ⅳ, HR 1.68(1.14–2.47), p=0.009. According to Table 3, DLL1 and DLL3 mRNA expressions were tied to worsening OS in intestinal, HR 1.6 (1.02–2.57), p = 0.048. HR 1.88 (1.34–2.64), p = 0.0002. DLL4 expression was so clearly tied to worsening OS in diffuse, HR 1.5 (1.06–2.13), p = 0.021. High expression of JAG1 was linked to improving OS in intestinal, HR 0.63 (0.46–0.86), p = 0.004 and mixed, HR 0.16 (0.03–0.73), p = 0.008. JAG2 mRNA expression was so clearly tied to worsening OS in intestinal, HR 2.72 (1.97–3.74), p = 1.8e-10 as well as diffuse, HR 1.66 (1.16–2.38), p = 0.005. According to Table-4, DLL3 expression was tied to a better OS in low differentiation adenocarcinoma, HR 0.61 (0.39–0.94), p = 0.024. However, with worsen OS in high differentiation adenocarcinoma, HR 2.57 (1.03–6.39), p = 0.036. JAG1 expression was tied to a better OS in low differentiation adenocarcinoma, HR 0.59 (0.38–0.91), p = 0.017 and high differentiation adenocarcinoma, HR 0.18 (0.04–0.79), p = 0.011. JAG2 mRNA expression was tied to worsening OS in high differentiation adenocarcinoma, HR 3.66 (1.32–10.15), p = 0.008. From Table 5, DLL3 mRNA expression was tied to worsening OS in surgery combined with 5-FU chemotherapy, HR 1.86 (1.30–2.67), p = 0.0006. DLL4 and JAG2 mRNA expression was so clearly tied to worsening OS in Surgery, same as surgery combined with 5-FU chemotherapy, p<0.05. JAG1 expression was just tied to worsening OS in Surgery, HR 1.35 (1.01–1.8), p = 0.042.