Introduction
Materials and methods
Samples and clinicopathological data
RNA extraction and quantitative real-time reverse transcriptase PCR (qRT-PCR)
Protein extraction and Western blot
Tissue microarray and immunohistochemistry
Assessment of immunohistochemistry staining
Statistical analysis
Results
Nodal and YAP1 mRNA expression are significantly up-regulated in GAC
Groups |
n
| Nodal expression |
P
| YAP1 expression |
P
|
---|---|---|---|---|---|
mRNA |
0.046
|
0.013
| |||
GC | 20 | 0.126 ± 0.177 | 0.105 ± 0.069 | ||
Normal | 20 | 0.056 ± 0.063 | 0.075 ± 0.052 | ||
Protein |
0.017
|
0.038
| |||
GC | 32 | 1.0699 ± 0.9241 | 0.8494 ± 0.2893 | ||
Normal | 32 | 0.6929 ± 0.2965 | 0.7012 ± 0.3667 |
Nodal and YAP1 protein expression are significantly up-regulated in GAC
Expression of Nodal and YAP1 are not correlated in GAC
High Nodal protein expression is associated with poor clinicopathological variables in GAC
Variable |
n
| Nodal expression | HPR (%) |
χ
2
|
P value | |
---|---|---|---|---|---|---|
Low | High | |||||
Age (year) | 3.696 | 0.055 | ||||
≤55 | 82 | 43 | 39 | 47.6 | ||
>55 | 138 | 54 | 84 | 60.9 | ||
Gender | 0.018 | 0.892 | ||||
Female | 67 | 30 | 37 | 55.2 | ||
Male | 153 | 67 | 85 | 56.2 | ||
Borrmann’s types | 0.26 | 0.61 | ||||
I + II | 30 | 12 | 18 | 60.0 | ||
III + IV | 189 | 85 | 104 | 55.0 | ||
WHO’s histology types | 8.930 | 0.159 | ||||
Papillary. Ade. | 8 | 3 | 5 | 62.5 | ||
Tubular Ade. | ||||||
Well Diff. | 14 | 9 | 5 | 35.7 | ||
Moderately Diff. | 74 | 36 | 38 | 51.4 | ||
Poorly Diff. | 99 | 35 | 64 | 64.6 | ||
Un-differentiated car. | 3 | 1 | 2 | 66.7 | ||
Signet ring cell Car. | 7 | 5 | 2 | 28.6 | ||
Mucinous Ade. | 15 | 8 | 7 | 46.7 | ||
Lauren’s types | 0.735 | 0.692 | ||||
Intestinal | 80 | 38 | 42 | 53.5 | ||
Diffuse | 102 | 42 | 60 | 58.8 | ||
Mixed | 38 | 17 | 21 | 55.3 | ||
Depth of invasion | 8.899 |
0.003
| ||||
T1 + T2 | 36 | 24 | 12 | 33.3 | ||
T3 + T4 | 184 | 73 | 111 | 60.3 | ||
Ln metastasis | 19.669 |
<0.001
| ||||
N0 | 60 | 41 | 19 | 31.7 | ||
N1–3 | 160 | 56 | 104 | 65.0 | ||
Distant metastasis | 23.815 |
<0.001
| ||||
M0 | 167 | 89 | 78 | 46.7 | ||
M1 | 53 | 8 | 45 | 84.9 | ||
TNM staging | 26.416 |
<0.001
| ||||
I + II | 71 | 49 | 22 | 31 | ||
III + IV | 149 | 48 | 101 | 67.8 | ||
YAP1 expression | 1.8712 | 0.171 | ||||
Low | 41 | 22 | 19 | 46.3 | ||
High | 179 | 75 | 104 | 58.1 |
High Nodal expression is associated with poor prognosis in GAC
Variable | Univariate analysisa
| Multivariate analysisb
| |||
---|---|---|---|---|---|
n
| Mean survival (months) |
P value | HR (95 % CI) |
P value | |
Borrmann’s types | 0.289 | 1.113 (0.646–1.985) | 0.663 | ||
I + II | 22 | 29.55 | |||
III + IV | 139 | 32.62 | |||
WHO’s histology types | 0.638 | 1.096 (0.913–1.317) | 0.324 | ||
Papillary. Ade. | 6 | 32.50 | |||
Tubular Ade. | |||||
Well Diff. | 11 | 33.80 | |||
Moderately Diff. | 56 | 35.29 | |||
Poorly Diff. | 70 | 29.71 | |||
Un-differentiated car. | 3 | 35.67 | |||
Signet ring cell car. | 5 | 16.60 | |||
Mucinous Ade. | 10 | 26.07 | |||
Lauren’s types | 0.354 | 1.055 (0.747–1.492) | 0.760 | ||
Intestinal | 56 | 35.11 | |||
Diffuse | 77 | 31.41 | |||
Mixed | 28 | 28.13 | |||
Depth of invasion | 0.001 | 1.886 (0.895–3.975) | 0.095 | ||
T1 + T2 | 28 | 43.96 | |||
T3 + T4 | 133 | 29.54 | |||
Ln metastasis | 0.001 | 0.450 (0.208–0.974) | 0.043 | ||
N0 | 43 | 41.06 | |||
N1–3 | 118 | 28.64 | |||
TNM staging | <0.001 | 6.999 (2.754–17.786) | <0.001 | ||
I + II | 52 | 46.15 | |||
III + IV | 109 | 25.23 | |||
Nodal protein expression | <0.001 | 2.487 (1.546–4.001) | <0.001 | ||
Low | 80 | 40.90 | |||
High | 81 | 23.58 | |||
YAP1 protein expression | 0.001 | 2.506 (1.195–5.257) | 0.015 | ||
Low | 29 | 44.90 | |||
High | 132 | 29.22 | |||
Nodal and YAP1 expression | <0.001 | ||||
Nodal−YAP1−
| 18 | 50.30 | |||
Nodal−YAP1+
| 59 | 37.35 | |||
Nodal+YAP1−
| 14 | 39.38 | |||
Nodal+YAP1+
| 70 | 21.51 |