Overall survival of patients were estimated by the Kaplan-Meier method, differences between groups were compared were by

the log-rank test. Multivariate analysis was performed using a Cox proportional hazard model. Statistically significant prognostic factors identified by univariate analysis were entered in the multivariate analysis. #LY3023414 in vivo randurls[1|1|,|CHEM1|]# All the statistical analyses were performed with SPSS 16.0 software. P value less than or equal to 0.05 was considered statistically significant. Results Expression of MAGE-A1, MAGE-A3/4, NY-ESO-1 and HLA class I proteins in IHCC patients by immunohistochemistry MAGE-A1, MAGE-A3/4 and NY-ESO-1 showed a predominantly, although not exclusively, cytoplasmic staining (Figure 1). The frequency and grade of various CTA expressions in tumors is shown in Table 1. Figure 2 showed a Venn diagram dipicting the overlap

of three CTAs expression. When the CTA combinations were tested, 52 from 89 IHCC cases (58.4%) showed expression of at least one marker, 14 cases (15.7%) demonstrated BMN 673 molecular weight co-expression of two CTAs, and only three cases (3.3%) were positive for all the three antigens. As seen in table 2, down-regulated HLA class I expression was found in 42.7% of all tumors (n = 38). Comparing the relationship between individual or combined CTAs expression and HLA-class I expression, no correlation was observed. And 30 IHCC cases (33.7%) demonstrated concomitant expression of CTAs and HLA class I antigen. Figure 1 Immunohistochemical analysis of MAGE-A1, MAGEA3/4, NY-ESO-1 and HLA Class I in intrahepatic

cholagiocarcinoma. Sections were stained with antibody against (A) MAGE-A1 (MA454); (B) MAGE-A3/A4 (57B); (C) NY-ESO-1 (E978); (D) HLA Class I (EMR8-5). Figure 2 Venn diagram depicting the overlap in the expression of cancer-testis antigens in intrahepatic cholagiocarcinoma. Table 1 Expression of cancer-testis antigens in intrahepatic cholanglocarcinoma   MAGE-A1 Interleukin-2 receptor N (%) MAGE-A3/4 N (%) NY-ESO-1 N (%) Negative 63 (70.8) 65 (73.0) 70 (78.7) Positive 26 (29.2) 24 (27.1) 19 (21.3)    + 2 (2.2) 1 (1.1) 1 (1.1)    ++ 3 (3.4) 4 (4.4) 1 (1.1)    +++ 12 (13.5) 14 (15.7) 7 (7.9)    ++++ 9 (10.1) 5 (5.6) 10 (11.2) Table 2 Correlation between CTA expression pattern and HLA class I expression CTA expression pattern HLA class I expression P value   Positive (n = 51) Down-regulated (n = 38)   MAGE-A1          Positive 18 8 0.144    Negative 33 30   MAGE-A3/4          Positive 11 13 0.184    Negative 40 25   NY-ESO-1          Positive 11 8 0.953    Negative 40 30   1 CTA positive          With 30 22 0.930    Without 21 16   2 CTA positive          With 9 5 0.565    Without 42 33   3 CTA positive          With 1 2 0.