These results are consistent with a number of previous reports. In the present study, the protein and mRNA levels of UPK1A in GC patients were evaluated by western blo ing and RT-PCR, respectively. The correlation of UPK1A and clinical outcome was analyzed by using immunohistochemical staining of primary 
GC tissue. The results revealed that the protein levels of UPK1A were significantly reduced in tumor tissue samples, compared with levels in normal paracancerous tissues. The RT-qPCR findings were consistent with western blo ing detections. These results support the previous hypothesis that UPK1A might be a tumor suppressor candidate in gastric cancer. Subsequently, using immunohistochemistry analysis for paraffin specimens, with large series of gastric cancer patients, we observed that the low UPK1A expression was associated with histological grade, lymph node metastases and UICC stage. These findings are similar to a previous study that reported an association between low UPK1A expression and lymph node metastasis as well as in esophageal squamous cell carcinoma. Similar findings can be observed in other malignancies. Therefore, UPK1A may serve as a potential tumor-suppressor gene in gastric cancer. In analyses focused on the survival of patients, the expression level of UPK1A was shown to be a significant predictive variable in surgically resected GC. In the Kaplan�CMeier survival analysis, the median OS was significantly longer among patients with high UPK1A expression than among those with low UPK1A expression. Furthermore, in the univariate analysis, UPK1A is a significant factor. However, in the multivariate analysis, UPK1A is an independent risk factor at the 0.1 level. Age, tumor location, tumor emboli in the vasculature, and TNM stage were independent risk factors in the prognosis of gastric cancer patients at 0.05 level. The findings may suggest that decreased UPK1A expression might help identify gastric cancer patients with poor prognosis and more lymph node metastases. However, not all of our findings have been reported elsewhere, and we cannot make a Procyanidin-B2 general statement concerning the influence of UPK1A expression on the lymph node metastases of GC. UPK1A holds b-catenin in the membrane. Therefore, by interacting with kinases, b-catenin is maintained at a low level. If glycogen synthase kinase 3 is inhibited, b-catenin will translocate to nucleus and cause carcinogenesis. Kar confirmed the translocation of b-catenin by testing the expression of its downstream targets, including MMP7, c-Jun, c-myc, and nD1cyclin, all of which were down-regulated. In vitro assays were used to analyze the function of UPK1A. Overexpression of UPK1A inhibited the migration and invasion of MKN45 GC cell line, which was consistent with our clinical data analysis. The down-regulation of UPK1A was significantly associated with lymph node metastasis in GC. Cell cycle analysis revealed that the overexpression of UPK1A led to cell cycle arrest at the G1-S checkpoint. These findings Ergosterol strengthen the hypothesis that UPK1A might play an important role in GC tumor suppression. Furthermore, consistent with our results, a previous study demonstrated that UPK1A could induce cell cycle arrest at the G1-S checkpoint.