To clarify the significance of circulating tumor cells (CTC) undergoing epithelial-mesenchymal transition (EMT) in patients with hepatocellular carcinoma (HCC), we used an advanced CanPatrol CTC-enrichment technique and hybridization to enrich and classify CTC from blood samples. One hundred and one of 112 (90.18%) patients with HCC were CTC positive, even with early-stage disease. CTCs were also detected in 2 of 12 patients with hepatitis B virus (HBV), both of whom had small HCC tumors detected within 5 months. CTC count ≥16 and mesenchymal-CTC (M-CTC) percentage ≥2% prior to resection were significantly associated with early recurrence, multi-intrahepatic recurrence, and lung metastasis. Postoperative CTC monitoring in 10 patients found that most had an increased CTC count and M-CTC percentage before clinically detectable recurrence nodules appeared. Analysis of HCC with high CTC count and high M-CTC percentage identified 67 differentially expressed cancer-related genes involved in cancer-related biological pathways (e.g., cell adhesion and migration, tumor angiogenesis, and apoptosis). One of the identified genes, BCAT1, was significantly upregulated, and knockdown in Hepg2, Hep3B, and Huh7 cells reduced cell proliferation, migration, and invasion while promoting apoptosis. A concomitant increase in epithelial marker expression (EpCAM and E-cadherin) and reduced mesenchymal marker expression (vimentin and Twist) suggest that BCAT1 may trigger the EMT process. Overall, CTCs were highly correlated with HCC characteristics, representing a novel marker for early diagnosis and a prognostic factor for early recurrence. BCAT1 overexpression may induce CTC release by triggering EMT and may be an important biomarker of HCC metastasis. In liver cancer, CTC examination may represent an important "liquid biopsy" tool to detect both early disease and recurrent or metastatic disease, providing cues for early intervention or adjuvant therapy. .

P53 mutations were detected in 223 HCC samples, 13 adjacent liver tissue samples and only 1 of 68 normal liver tissue samples. The mutation sites concentrated at exon 4, 5, 6, 7, 8, 9 and no mutation was detected in exon 1, 2, 3, 10 and 11. The most frequently occurring mutation was in codon 249 (R249S) in exon 7. Patients in the HBV(+)/AFB1(+) and HBV(-)/AFB1(+) groups had significantly higher mutation rates compared with patients in the HBV(+)/AFB1(-) and HBV(-)/AFB1(-) groups. P53 mutation status and HBV/AFB1 status were independent predictors of tumour recurrence after surgery. Immunohistochemical analysis revealed that p53 gene mutations were correlated with the p53 expression. In Guangxi area, the significant association between AFB1-induced p53 mutations and the expression of p53 protein suggest an important role for p53 mutations in carcinogenesis of HCC.

Background and aims: Hepatic resection is the first-line treatment for hepatocellular carcinoma (HCC). Whether to perform anatomical (AR) or non-anatomical resection (NAR) remains controversial. This retrospective study compares the outcomes according to the number and type of circulating tumour cells (CTCs). Methods: The cohort included 136 patients with HCC treated with R0 resection between 2014 and 2017. CanPatrol CTC-enrichment technique was used to enrich and classify CTCs according to epithelial-to-mesenchymal transition phenotype. Results: 91.91% of total patients were CTC-positive, with 91.23% in the AR group and 92.41% in the NAR group. Tumour-free survival (TFS) did not differ significantly between the two groups. However, TFS was significantly higher in patients with low CTCs count and mesenchymal-and epithelial/mesenchymal-negative phenotypes. As for the incidence and types of recurrence, high pre-resection CTC count and mesenchymal-and epithelial/mesenchymal-positivity were significantly associated with extrahepatic and multi-intrahepatic recurrence. Higher morbidities for hepatic failure and ascites were observed in patients treated by AR. Conclusion: AR may be more beneficial than NAR only in patients with low CTC count and mesenchymal-and epithelial/mesenchymal-negative phenotypes. For patients with a high CTC count, the balance between operative risk and prognostic benefit is more important than the resection method performed. KEY MESSAGESAnatomic resection may improve the survival of HCC patients, but only those with low CTC count and negative M-and E/M-CTC phenotypes. CTC analysis before surgery can be used to better guide the choice of resection method for HCC.

BackgroundThe aim of this study was to investigate the long-term survival and prognosis for primary clear cell carcinoma of the liver (PCCCL) of the liver after hepatectomy.MethodsOur study retrospectively analyzed the clinicopathological data of 64 patients with PCCCL (PCCCL group) and 247 with nonclear cell hepatocellular carcinoma (NHCC group) after hepatectomy between January 1996 and December 2006. The overall survival (OS) and disease-free survival of the two groups was compared using the Kaplan–Meier method. Prognostic factors of survival were identified by multivariate analysis.ResultsThe 1-, 3-, and 5-year OS (P=0.016) and disease-free survival (P<0.001) of the PCCCL group were significantly higher than that of the NHCC group. In mutivariate analysis, tumor size >5 cm, presence of portal vein tumor thrombosis and proportion of clear cells ≤70% were risk factors for OS of the PCCCL group. The prognosis of a subgroup with higher proportion of clear cells was markedly better than that of the subgroup with a lower proportion of clear cells.ConclusionOur results suggested that the prognosis of patients with PCCCL was better than that of the patients with NHCC. The higher the proportion of clear cells, the better the prognosis.

ObjectiveThis study aimed to explore the relationship between KI-67 positive cellular index and recurrence-free survival (RFS) in Barcelona Clinic Liver Cancer (BCLC) stage A and B hepatocellular carcinoma (HCC) patients, particularly those with microvascular invasion (MVI).MethodsA total of 333 patients who underwent curative hepatectomy had their immunohistochemistry analyzed retrospectively for KI-67 positive cellular index.ResultsIn total, 41.1% (137/333) of HCC patients displayed high KI-67 positive cellular index (>35%). Patients with high KI-67 positive cellular index had poorer RFS than those with low index (P<0.0001). Patients were then subdivided into an MVI positivity group (n=192) and an MVI negativity group (n=141). In the MVI positivity group, patients with high KI-67 positive cellular index had a shorter RFS after operation as compared to those with low index (P<0.0001). However, there was no significant difference in RFS between high- and low-index subgroups within the MVI negativity group (P>0.05). Additionally, patients with high KI-67 positive cellular index combined with MVI positivity had the shortest RFS of all those with MVI negativity, regardless of KI-67 cellular index level (P<0.0001). Multivariate analysis showed that node number >1, capsule absence, high KI-67 positive cellular index, and alpha-fetoprotein >400 ng/mL were independent risk factors for a recurrence of HCC with MVI.ConclusionOur results suggested that high KI-67 positive cellular index may represent a poor prognostic factor in BCLC stage A and B HCC patients, especially those with MVI.