015), and ezrin (P = 0.092) positivity and loss of E-cadherin expression (P = 0.006), compared to CD133-negative HCCs. Ezrin expression was more common in EpCAM-positive HCCs, compared to EpCAM-negative HCCs (P = 0.007), and snail was more frequently expressed in c-kit-positive HCCs, compared to c-kit-negative HCCs (P = 0.031). A univariable survival analysis,

according to the expression status of the four stemness-related markers, demonstrated that K19 expression in HCC was associated with a poor overall (P = 0.018) and disease-free survival (P = 0.007) (Supporting Table 3). CD133-expressing HCCs showed decreased overall survival (P = 0.057), compared to CD133-negative HCCs, although this website marginally significant. EpCAM or c-kit expression status was not related to HCC prognosis in this study. In addition, expression of three or more stemness-related proteins in HCC was characterized by a decreased overall survival (P = 0.086); however, selleck disease-free

survival was not affected by this variable. Because the majority (n = 101) of cohort 1 HCCs were HBV-related, survival analysis was repeated separately for 101 HBV-related HCCs and 36 HBV-unrelated HCCs (11 HCV-related, 6 alcohol-related, and 19 of uncertain etiology) to see whether K19 would be still prognostically significant in non-B-viral HCCs. Disease-free survival was still significantly decreased in K19-positive, HBV-unrelated HCCs, compared to K19-negative cases (P = 0.005) (Supporting Fig. 3). Overall survival, however, was not significantly different Cepharanthine between the two groups. As for the HBV-related HCCs, decreased overall survival (P = 0.002) and disease-free survival (P = 0.121) were noted in the K19-positive groups. Because K19 appeared to be the stemness-related marker that was most significantly associated with clinicopathologic

features of tumor aggressiveness, the next cohort was divided into two groups, according to K19 protein expression status. Immunohistochemical stain for K19 protein was done using whole sections of representative paraffin blocks from each case, and K19 positivity was found in 68 (28.7%) cases. The pattern of K19 immunoreactivity was focal and heterogeneous. The smaller tumor cells at the periphery of the tumor-cell nests, or intermingled with the hepatocyte-like cells within the tumor-cell nests, expressed K19 in 16 cases, whereas the majority of the K19-expressing tumor cells were hepatocyte-like cells (n = 52). HCCs were grouped according to K19 protein-expression status, and clinicopathological features were compared between the two groups (Table 2). The K19-positive group was composed of higher proportions of younger (P = 0.004) and female (P < 0.001) patients, compared to the K19-negative group. K19-positive HCCs were more frequently associated with high AFP levels (>1,000 IU/mL), compared to K19-negative HCCs (P < 0.001). On pathologic examination, K19-positive HCCs showed more frequent microvascular invasion (P = 0.017) and fibrous stroma (P = 0.