Univariate and multivariate Cox regression were used to identify risk factors for prediction of recurrence and death in patients with HCC after initial curative hepatectomy. www.selleckchem.com/products/bay80-6946.html In the univariate analysis, the effect of each variable on the disease recurrence and survival was tested using analysis of variance. If there was a known predominant determinant (e.g., tumor stage versus vascular invasion), bivariate analysis was conducted to detect more detailed correlations independent from the predominant one that might have been unnoticed. Only variables with a P value less than 0.05 were selected for subsequent multivariate analysis. In multivariate analysis, the

selected variables were subjected to a multiple linear model. Hazard ratios (HR) with 95% confidence intervals (CI) were thus obtained. Coefficients were determined via the linear discriminating function of the variables. To examine the potential role of NPM in resistance to anticancer therapies of HCC, HCC cell

lines with different p53 genetic background including HepG2 (wild-type p53), Huh7 (C200Y mutated p53), Mahlavu selleck (R249S mutated p53), and Hep3B (null-genotyped p53),23, 24 were treated with UV-B, mitomycin C, doxorubicin, or cisplatin. Silencing of NPM expression significantly enhanced cellular susceptibility to all MCE kinds of treatments in Huh7, Hep3B, and Mahlavu cells, while the sensitizing effect was minimal in HepG2 cells (Fig. 1A). These findings suggest that an NPM-mediated antideath mechanism is independent of p53 functions in HCC cells, which is different from that found in hematopoietic cells.25 To further inspect the role of p53, we silenced the expression of NPM, p53, or simultaneously NPM and p53 by RNA interference (Fig. 1B). Silencing of p53 expression alone did not significantly change the sensitivity to any of the treatments in Huh7, Hep3B, and Mahlavu cells (Fig. 1B, siTP53 versus siNS). Simultaneous silencing of p53 and NPM did not further

alter the sensitizing effect exerted by silencing of NPM alone (Fig. 1B, siNPM versus [siNPM + siTP53]). Silencing of NPM and p53 expression by RNA interference was confirmed via immunoblotting (Fig. 1C). Interestingly, a negative dominant mutant of p53 converted HepG2 cells from insensitivity to sensitivity to cytotoxic and molecular targeted therapies as NPM silenced (Supporting Fig. 1). NPM apparently executes its death evasion activity independently of p53 function. We further examined the potential role of NPM in resistance to the inhibitors of oncogenic kinases in HCC, such as lapatinib and sorafenib. Silencing of NPM expression significantly sensitized Huh7, Hep3B, and Mahlavu cells to sorafenib and lapatinib (Fig. 2A).