A liver tumor was detected in a 72 year old male patient with liver cirrhosis Child-Pugh Stage A, a history of type 2 diabetes and chronic hepatitis C virus infection (HCV, genotype 1B), initially diagnosed 13 years ago. Liver enzymes were slightly elevated with alanine aminotransferase 89 U/L (reference: 10–50 U/L) and aspartate aminotransferase 65 U/L (reference: <50 U/L). The 4 cm tumor was detected by routine sonography and removed by laparoscopic liver segment resection. Morphological analysis, immunohistochemistry and multiregional, next generation sequencing (NGS) was applied on representative tumor sections as described []. Table and Fig. illustrate histopathological and molecular findings in 14 individual tumor areas, which were grouped into three tumor regions (A, B and C) according to their predominant morphological and molecular characteristics. In summary, a multinodular, combined hepatocellular/cholangiocellular carcinoma, tumor stage T1 grade 2–3, was diagnosed. Intratumoral heterogeneous expression of five liver cell markers (CK7, CK19, glutamine synthetase, p53, β-catenin) was detected including a double positivity for glutamine synthetase, nuclear β-catenin and p53 in tumor region A. Next generation sequencing was performed with a minimum coverage of 1329 (SD ± 725) reads per amplicon of every single tumor area. Sequencing results yielded a p.D32V (c.363 A > T) mutation and a TP53 ivs8-1 (c.783-1 G > A) splice site mutation in tumor region A. Comparing mutated allele frequencies, CTNNB1 and TP53 gene copies showed a similar range of both frequencies in area A1-3. All mutated and wild type tumor areas additionally displayed a SNP of exon 7 (rs17880604). Collectively, morphological and immunohistochemical findings together with sequencing results demonstrated that a tumor subclone with hepatocellular differentiation had concomitant CTNNB1 and TP53 gene mutations. To date, after a follow-up time of 12 months, the patient had a local recurrence of a liver tumor which was inoperable and therefore treated by transarterial chemoembolization (TACE).