Sineoculis homeobox homolog 1 protein as an independent biomarker for gastric adenocarcinoma.

ABSTRACT

Sine oculis homeobox homolog 1 (SIX1) protein is a member of the homeobox transcription factor family. Overexpression of SIX1 contributes to cancer progression and is associated with adverse outcomes in various cancer types including breast, ovarian, uterine cervical and liver. To investigate the clinicopathological significance of SIX1 protein expression in gastric adenocarcinomas (GAC), localization of the SIX1 protein was determined in MKN-1, a gastric cancer cell line, using immunofluorescence (IF) staining; SIX1 mRNA level was detected in fresh tissues of GAC and normal gastric mucosa using quantitative real-time polymerase chain reaction (qRT-PCR); and SIX1 protein expression was assessed in 163 GAC, 35 gastric dysplasia and 26 normal gastric mucosa using immunohistochemical (IHC) staining. Correlations between SIX1 protein expression and pathological parameters of GAC were analyzed using Chi-square tests, differences in survival curves were analyzed using log-rank tests, and multivariate survival analysis was performed using the Cox proportional hazards regression model. SIX1 protein showed a mainly cytoplasmic staining pattern in GAC using IF and IHC staining. The positive SIX1 protein expression rate was 80.4% in GAC, which was significantly higher than in either gastric dysplasia (45.7%) or normal gastric mucosa (26.9%) (P<0.01). qRT-PCR data also confirmed increased levels of SIX1 mRNA expression in GAC compared with the normal gastric mucosa in fresh tissues. In addition, the strongly positive SIX1 protein expression rate was significantly correlated with clinical stage, lymph node metastasis and serosal invasion of GAC (P<0.01 or P<0.05), while there was no association with gender, age, tumor size, Lauren classification or histological types of GAC. Notably, strongly positive signals were frequently observed in tumor blood vessels and/or lymphatic vessels. GAC patients with high expression of the SIX1 had shorter overall and disease-free survival rates than those with low SIX1 protein expression (P<0.01). Furthermore, using multivariate analysis, SIX1 protein expression was found to be an independent risk factor for survival in patients with GAC along with clinical stage and serosal invasion (P<0.01). In conclusion, SIX1 protein expression status may be an independent biomarker for prognostic evaluation of GAC.