Malvi D, de Biase D, Fittipaldi S, et al. - A total of 28 mixed primary mixed liver cancers (PLCs), 13 combined hepatocellular-cholangiocellular (cHCC-CC) and 15intermediate-cell carcinomas were chosen in order to ascertain the expression of Nestin in mixed PLCs and to equate the PLC immunoprofile with the gene expression in each tumor component. For Nestin, in all cases, the differentiated HCC and CC components of cHCC-CC were negative. The intermediate areas of cHCC-CC were immunoreactive for Nestin, CD56, and K7/K19 in 92.3%, 76.9% and for in all cases, respectively. The immunoprofile of the intermediate-cell carcinomas exhibited 73.3% and 66.7% of cases positive for Nestin and for CD56, respectively. The most commonly mutated genes were TP53 and TERT. Also in IDH1, IDH2, PIK3CA and NRAS genes, mutations were seen. Intermediate and HCC areas of cHCC-CC appeared to share the same mutational profile, and both harbored various mutations compared with the CC component. In conclusion, Nestin, according to the preliminary data, was not expressed by hepatocellular or cholangiocellular-cell components, however, it was expressed through most of the intermediate cells in PLCs, and hence, could be regarded in the differential diagnosis of PLCs, collectively with the mutational profile.