[PubMed] [Google Scholar] 7. expression group (0.1< IHC score <0.9, n=32) and the very weak CLIC1 expression group (IHC score 0.1, n=20). The 5-year overall survival rate of the very strong CLIC1 expression group CP 945598 HCl (Otenabant HCl) and that of the very weak CLIC1 expression group were significantly poorer than that of the middle CLIC1 expression group (Supplementary Physique 3). We investigated whether the very strong or very weak expression of CLIC1 was prognostic for ESCC patients Rabbit Polyclonal to FPR1 after curative resection. The univariate analysis showed that the presence of lymphatic invasion, venous invasion, and the pathological depth of the tumor correlated with a poor 5-year overall survival rate. The 5-year overall survival rate of the very strong or very weak CLIC1 expression group was 44.8%, which was significantly poorer than that of the other group (84.2%) (p=0.001). A multivariate analysis with these three factors and an IHC score 0.9 or 0.1 revealed that the very strong or very weak expression of CLIC1 was an independent prognostic factor (Table ?(Table4).4). These results suggest that very strong or very weak expression of CLIC1 in ESCC tissues is related to the poor prognosis of patients with ESCC after curative resection. Table 4 Five-year overall survival rates of patients with ESCC according to various clinicopathological parameters experiments with ESCC cells, the expression of CLIC1 regulated tumor behaviors, including cell proliferation, apoptosis, and cellular movement, and our immunohistochemical results supported those obtained in experiments; that is, the group of very strong CLIC1 expression was poorer prognosis due to inhibiting apoptosis of ESCC cells, and the group of very weak CLIC1 expression was poorer prognosis due to promoting cell movement of ESCC cells. In short, our results indicate that CLIC1 expression levels are related to the switching of the tumor behaviors of ESCC. Although a deeper understanding of CLIC1 expression and CP 945598 HCl (Otenabant HCl) its heterogeneity in biopsy specimen is needed, further analyses may be helpful in the clinical use of CLIC1 IHC score as a preoperative biomarker in future. In summary, we herein exhibited that CLIC1 plays a role in the proliferation, apoptosis, and cellular movement of ESCC cells. Our microarray data also showed that CLIC1 affects the expression of other CP 945598 HCl (Otenabant HCl) genes with functions related to cell proliferation and apoptosis. Immunohistochemistry revealed that the very strong or weak expression of CLIC1 in human ESCC tissue was related to the prognosis of ESCC patients. Although further investigations around the underlying molecular mechanisms are needed, the present results suggest that CLIC1 is usually a useful biomarker of tumor progression and/or a novel therapeutic target for the future treatment of ESCC. MATERIALS AND METHODS Cell lines, antibodies, and other reagents The poorly differentiated human ESCC cell lines, TE2, TE5, and TE9, moderately differentiated human ESCC cell line, TE8, and well-differentiated human ESCC cell line, TE15, were obtained from the Cell Resource Center of Biomedical Research Institute of Development, Aging, and Cancer (Tohoku University, Sendai, Japan). The poorly differentiated human CP 945598 HCl (Otenabant HCl) ESCC cell lines, KYSE70 and KYSE150, moderately differentiated human ESCC cell line, KYSE170, and well-differentiated human ESCC cell line, KYSE790, were obtained from Kyoto University (Kyoto, Japan). These cells were produced in RPMI-1640 medium (Nacalai Tesque, Kyoto, Japan) supplemented with 100 U/mL.