Prognostic value of small mother against decapentaplegic expression in human gastric cancer

Figures & data

Figure 1. Prognostic value of SMAD1 expression in gastric cancer. Prognostic value of SMAD1 expression as shown by KM plotter (www.kmplot.com) curves. Affymetrix ID for SMAD1: 227798_ at. OS curves were plotted for A: all the patients (n = 876); B: intestinal cancer patients; C: diffuse cancer patients; D: mixed cancer patients

Figure 2. Prognostic value of SMAD2 expression in gastric cancer

Prognostic value of SMAD2 expression as shown by KM plotter (www.kmplot.com) curves. Affymetrix ID for SMAD1: 203077_s_ at. OS curves were plotted for A: all the patients (n = 876); B: intestinal cancer patients; C: diffuse cancer patients; D: mixed cancer patients.

Figure 3. Prognostic value of SMAD3 expression in gastric cancer

Prognostic value of SMAD3 expression as shown by KM plotter (www.kmplot.com) curves. Affymetrix ID for SMAD3: 218284_ at. OS curves were plotted for A: all the patients (n = 876); B: intestinal cancer patients; C: diffuse cancer patients; D: mixed cancer patients.

Figure 4. Prognostic value of SMAD4 expression in gastric cancer

Prognostic value of SMAD4 expression as shown by KM plotter (www.kmplot.com) curves. Affymetrix ID for SMAD4: 202527 _ s _ at. OS curves were plotted for A: all the patients (n = 876); B: intestinal cancer patients; C: diffuse cancer patients; D: mixed cancer patients.

Figure 5. Prognostic value of SMAD5 expression in gastric cancer

Prognostic value of SMAD5 expression as shown by KM plotter (www.kmplot.com) curves. Affymetrix ID for SMAD5: 205188 _ s _ at. OS curves were plotted for A: all the patients (n = 876); B: intestinal cancer patients; C: diffuse cancer patients; D: mixed cancer patients.

Figure 6. Prognostic value of SMAD6 expression in gastric cancer

Prognostic value of SMAD6 expression as shown by KM plotter (www.kmplot.com) curves. Affymetrix ID for SMAD6: 213565 _ s _ at. OS curves were plotted for A: all the patients (n = 876); B: intestinal cancer patients; C: diffuse cancer patients; D: mixed cancer patients.

Figure 7. Prognostic value of SMAD7 expression in gastric cancer

Prognostic value of SMAD7 expression as shown by KM plotter (www.kmplot.com) curves. Affymetrix ID for SMAD7: 204790 _ at. OS curves were plotted for A: all the patients (n = 876); B: intestinal cancer patients; C: diffuse cancer patients; D: mixed cancer patients.

Figure 8. Prognostic value of SMAD9 expression in gastric cancer

Prognostic value of SMAD9 expression as shown by KM plotter (www.kmplot.com) curves. Affymetrix ID for SMAD9: 227719 _ at. OS curves were plotted for A: all the patients (n = 876); B: intestinal cancer patients; C: diffuse cancer patients; D: mixed cancer patient.

Table 1. Correlation of SMAD gene expression level with overall survival (OS) of gastric cancer patients in different pathological stages

SMADs	Clinic stages	Cases	HR (95% CI)	P-value
SMAD1	1	62	0.18 (0.06 − 0.59)	0.0016*
	2	135	0.48 (0.23 − 1.01)	0.048*
	3	197	0.59 (0.4 − 0.86)	0.0053*
	4	140	0.8 (0.54 − 1.18)	0.25
SMAD2	1	67	0.22 (0.07 − 0.69)	0.0046*
	2	140	0.52 (0.23 − 1.17)	0.11
	3	305	0.4 (0.28 − 0.59)	1.6e−06*
	4	148	0.71 (0.47 − 1.09)	0.12
SMAD3	1	67	0.42 (0.15 − 1.15)	0.081
	2	140	2.24 (1.04 − 4.81)	0.035*
	3	305	1.72 (1.29 − 2.29)	0.00017*
	4	148	0.68 (0.46 − 1)	0.047*
SMAD4	1	67	0.34 (0.12 − 0.99)	0.039*
	2	140	0.71 (0.37 − 1.35)	0.29
	3	305	0.42 (0.3 − 0.6)	8e−07*
	4	148	0.83 (0.55 − 1.25)	0.37
SMAD5	1	67	5.43 (0.71 − 41.71)	0.068
	2	140	2.64 (1.44 − 4.84)	0.0011*
	3	305	1.74 (1.24 − 2.45)	0.0012*
	4	148	2 (1.31 − 3.05)	0.001*
SMAD6	1	67	1.66 (0.53 − 5.17)	0.38
	2	140	2.55 (1.18 − 5.49)	0.013*
	3	305	1.4 (1.05 − 1.86)	0.022*
	4	148	1.4 (0.95 − 2.05)	0.084
SMAD7	1	67	3.4 (1.25 − 9.25)	0.011*
	2	140	1.87 (1.02 − 3.44)	0.04*
	3	305	1.35 (1.01 − 1.8)	0.04*
	4	148	0.76 (0.51 − 1.14)	0.19
SMAD9	1	62	4.42 (0.97 − 20.11)	0.036*
	2	135	2.17 (1.03 − 4.56)	0.037*
	3	197	1.44 (0.97 − 2.15)	0.067
	4	140	1.61 (1.08 − 2.39)	0.019*

*P < 0.05

Abbreviations: CI, confidence interval; HR, hazard ratio.

Table 2. Correlation of SMAD gene expression level with overall survival (OS) of gastric cancer patients with different differentiation

SMADs	Differentiation	Cases	HR (95% CI)	P-value
SMAD1	Poor	121	1.51 (0.91 − 2.5)	0.11
	Moderate	67	0.5 (0.25 − 1.01)	0.048*
	Well	32	Not available	Not available
SMAD2	Poor	165	1.22 (0.81 − 1.85)	0.34
	Moderate	67	0.59 (0.29 − 1.18)	0.13
	Well	32	0.6 (0.25 − 1.42)	0.24
SMAD3	Poor	165	1.26 (0.82 − 1.95)	0.29
	Moderate	67	2.45 (1.22 − 4.93)	0.0099*
	Well	32	3.97 (0.92 − 17.13)	0.047*
SMAD4	Poor	165	0.81 (0.53 − 1.24)	0.34
	Moderate	67	0.5 (0.25 − 1.02)	0.054
	Well	32	0.35 (0.14 − 0.89)	0.021*
SMAD5	Poor	165	0.81 (0.55 − 1.21)	0.31
	Moderate	67	1.74 (0.8 − 3.82)	0.16
	Well	32	0.47 (0.19 − 1.15)	0.09
SMAD6	Poor	165	1.64 (1.1 − 2.44)	0.014*
	Moderate	67	0.6 (0.28 − 1.28)	0.18
	Well	32	0.55 (0.23 − 1.31)	0.17
SMAD7	Poor	165	1.62 (1.08 − 2.45)	0.019*
	Moderate	67	2.08 (0.85 − 5.13)	0.1
	Well	32	2.72 (0.8 − 9.27)	0.097
SMAD9	Poor	121	1.42 (0.82 − 2.43)	0.21
	Moderate	67	1.81 (0.79 − 4.14)	0.15
	Well	32	Not available	Not available

*P < 0.05

Abbreviations: CI, confidence interval; HR, hazard ratio.

Table 3. Correlation of SMAD gene expression level with overall survival (OS) of gastric cancer patients of different genders

SMADs	Gender	Cases	HR (95% CI)	P-value
SMAD1	Female	187	0.39 (0.21 − 0.7)	0.0011*
	Male	349	0.65 (0.48 − 0.87)	0.0038*
SMAD2	Female	236	0.53 (0.36 − 0.8)	0.0021*
	Male	545	0.48 (0.36 − 0.62)	3.9e−08*
SMAD3	Female	236	2.26 (1.57 − 3.25)	6.2e−06*
	Male	545	1.83 (1.47 − 2.27)	3.3e−08*
SMAD4	Female	236	0.62(0.44–0.88)	0.0075*
	Male	545	0.55 (0.43 − 0.71)	1.4e−06*
SMAD5	Female	236	1.27 (0.9 − 1.81)	0.17
	Male	415	1.41 (1.14 − 1.75)	0.0015*
SMAD6	Female	236	1.73 (1.21 − 2.48)	0.0023*
	Male	545	1.38 (1.1 − 1.73)	0.0053*
SMAD7	Female	236	1.64 (1.15 − 2.35)	0.0059*
	Male	495	1.17 (0.93 − 1.48)	0.17
SMAD9	Female	187	2.54 (1.38 − 4.67)	0.002*
	Male	349	1.66 (1.23 − 2.22)	0.00068*

*P < 0.05

Abbreviations: CI, confidence interval; HR, hazard ratio.

Table 4. Correlation of SMAD gene expression level with overall survival (OS) of gastric cancer patients under different treatment strategies

SMADs	Treatments	Cases	HR (95% CI)	P-value
SMAD1	Surgery alone	380	0.69 (0.49 − 0.97)	0.03*
	5-FU-based adjuvant	34	0.52 (0.21 − 1.31)	0.16
	Other adjuvant	76	0.5 (0.2 − 1.25)	0.13
SMAD2	Surgery alone	380	0.65 (0.48 − 0.89)	0.0068*
	5-FU-based adjuvant	153	1.31 (0.89 − 1.93)	0.16
	Other adjuvant	76	0.56 (0.23 − 1.38)	0.2
SMAD3	Surgery alone	380	1.41 (1 − 1.99)	0.051
	5-FU-based adjuvant	153	2.23 (1.46 − 3.42)	0.00015*
	Other adjuvant	76	1.85 (0.67 − 5.1)	0.23
SMAD4	Surgery alone	380	0.75 (0.56 − 1)	0.046*
	5-FU-based adjuvant	153	0.72 (0.49 − 1.04)	0.077
	Other adjuvant	76	0.45 (0.13 − 1.52)	0.18
SMAD5	Surgery alone	380	1.36 (0.97 − 1.91)	0.07
	5-FU-based adjuvant	153	0.38 (0.25 − 0.57)	1.3e−06*
	Other adjuvant	76	0.4 (0.15 − 1.11)	0.069
SMAD6	Surgery alone	380	1.17 (0.88 − 1.56)	0.28
	5-FU-based adjuvant	153	1.77 (1.23 − 2.55)	0.002*
	Other adjuvant	76	1.56 (0.65 − 3.78)	0.32
SMAD7	Surgery alone	380	1.64 (1.21 − 2.22)	0.0012*
	5-FU-based adjuvant	153	1.16 (0.82 − 1.63)	0.4
	Other adjuvant	76	1.8 (0.74 − 4.34)	0.19
SMAD9	Surgery alone	380	1.58 (1.17 − 2.14)	0.0029*
	5-FU-based adjuvant	34	1.67 (0.67 − 4.17)	0.27
	Other adjuvant	76	2.86 (1.14 − 7.19)	0.019*

*P < 0.05

Abbreviations: 5-FU, 5-Fluorouracil; CI, confidence interval; HR, hazard ratio.

Table 5. Correlation of SMAD gene expression level with overall survival (OS) of gastric cancer patients under different Her2 status

SMADs	Her 2 status	Cases	HR (95% CI)	P-value
SMAD1	Negative	429	0.48 (0.34 − 0.68	1.6e−05*
	Positive	202	0.66 (0.45 − 0.97)	0.035*
SMAD2	Negative	532	0.52 (0.4 − 0.66	1.5e−07*
	Positive	344	1.16 (0.89 − 1.5)	0.27
SMAD3	Negative	532	1.57 (1.22 − 2.03)	0.00039*
	Positive	344	1.73 (1.33 − 2.25)	3.5e−05*
SMAD4	Negative	532	0.64 (0.51 − 0.8)	0.00013*
	Positive	344	0.72 (0.55 − 0.93)	0.012*
SMAD5	Negative	532	1.37 (1.09 − 1.72)	0.0061*
	Positive	344	1.32 (0.99 − 1.75)	0.057
SMAD6	Negative	532	1.47 (1.17 − 1.84)	0.00071*
	Positive	344	1.44 (1.09 − 1.9)	0.01*
SMAD7	Negative	532	1.52 (1.2 − 1.92)	0.00044*
	Positive	344	0.8 (0.62 − 1.04)	0.089
SMAD9	Negative	429	1.85 (1.39 − 2.47)	2.1e−05*
	Positive	202	1.7 (1.16 − 2.51)	0.0064*

*P < 0.05

Abbreviations: HER2, Human Epidermal Growth Factor Receptor-2; CI, confidence interval; HR, hazard ratio.

Figure 9. (e)-SIS3 inhibits proliferation of gastric cancer cells

(A) Label-free Real-time Cellular Analysis (RTCA) of AGS and MGC803 cells incubated with (E)-SIS3 (0, 10, 20 μM). (B) The proliferation of AGS and MGC803 cells with or without (E)-SIS3 treatment analyzed by colony formation assay. (C) Ki67 Immunofluorescence following AGS and MGC803 cells incubated with (E)-SIS3 (0,10, and 20 μM). Scale bar = 25 μm. Data are presented as mean ± SD, N = 3; ****P < 0.0001, compared with control.

Figure 10. (e)-SIS3 inhibits the migration of gastric cancer cells

Migration of AGS (A and B) and MGC803 (C and D) cells incubated with 10 and 20 μM (E)-SIS3 or an equal volume of DMEM medium for 24 hours. (E)-SIS3 significantly inhibited migration of AGS and MGC803 gastric cancer cells. Scale bar = 500 μm. *P < 0.05; ***P < 0.001; ****P < 0.0001, compared with control.

Figure 11. (e)-SIS3 induces apoptosis of gastric cancer cells

Flow cytometry results for apoptosis [apoptosis ratio was calculated as (Q2+ Q3)/(Q1+ Q2 + Q3+ Q4)] of AGS (A and B) and MGC803 (C and D) cells incubated with 10 μM and 20 μM (E)-SIS3 or an equal volume of DMEM medium for 24 hours. (E)-SIS3 significantly induced apoptosis in AGS and MGC803 cells. (E)-SIS3 induced apoptosis in a concentration-dependent manner. Data are presented as mean ± SD, N = 5; *P < 0.05; ***P < 0.001; ****P < 0.0001, compared with control.

Figure 12. (e)-SIS3 inhibits proliferation and induces apoptosis of gastric cancer cells through influencing SMAD3 gene expression

(A) The expression changes of SMAD3 genes in the AGS cell of gastric cancer after treating with (E)-SIS3 for 24 hours. (B) Changes in expression of SMAD3 genes in the MGC803 cell after treatment with (E)-SIS3 for 24 hours. Data are presented as mean ± SD, N = 3; **P < 0.01; ***P < 0.001; ****P < 0.0001, compared with control.