Clinical implications and biological features of a novel postoperative recurrent HCC classification: A multi-centre study.

BACKGROUND & AIMS: The multiplicity of hepatocellular carcinoma (HCC) recurrence patterns is the most important determinant of patients' postsurgical survival. A systematic HCC recurrence classification is needed to help prevent and treat postoperative HCC recurrence in the era of precision medicine. METHODS: A total of 1319 patients with recurrent HCC from four hospitals were enrolled and divided into a development cohort (n = 916), internal validation cohort (n = 225) and external validation cohort (n = 178). A comprehensive study of patients' clinicopathological factors and biological features was conducted. RESULTS: Four subtypes of recurrence were identified, which integrated recurrence features, survival, effects on systemic and liver function and potential therapeutics after recurrence: type I (solitary-intrahepatic oligorecurrence); type II (multi-intrahepatic oligorecurrence); type III (progression recurrence) and type IV (hyper-progression recurrence). Type III~IV recurrence indicated exceptionally poor prognosis. Subsequently, two nomogram models were established for type III~IV recurrence prediction, and both demonstrated excellent predictive performance and applicability of pre and postoperative strategy formulation. Multiple biological analyses revealed that HCC cases with type III~IV recurrence were characterized by enrichment in p53 mutations, CCND1 amplification, high proliferation/metastasis potential, inactive metabolism and immune exhaustion features. Over-expression of high mobility group protein 2 (HMGA2) enhanced the highly malignant behaviour of HCC through multiple molecular pathways, making it a potential prognostic predictor and therapeutic target. CONCLUSIONS: This 'recurrent HCC classification' has important potential value in identifying patients with surgical benefit, predicting postsurgical survival and guiding treatment strategies. Multidimensional biological insights also increased knowledge of factors associated with HCC recurrence.

The p53 mutation spectrum in hepatocellular carcinoma from Guangxi, China : role of chronic hepatitis B virus infection and aflatoxin B1 exposure.

BACKGROUND & AIMS: p53 is one of the most frequently mutated human tumour suppressor genes. Chronic infection with hepatitis B virus (HBV) and exposure to aflatoxin B1 (AFB1) induces p53 mutations in hepatocellular carcinoma (HCC) tissue. The aims of present study are to investigate the p53 mutation spectrum in HBV- and AFB1-related hepatocarcinogenesis in patients with hepatocellular carcinoma (HCC) in Guangxi, China. METHODS: Tumour and adjacent liver tissue were collected from 397 HCC patients who were subdivided into HBV(+)/AFB1(+), HBV(+)/AFB1(-), HBV(-)/AFB1(+) and HBV(-)/AFB1(-) four groups. All 11 exons of the p53 gene were PCR-amplified and sequenced. Immunohistochemistry was used to evaluate the effect of mutations on the expression of p53 protein. RESULTS AND CONCLUSIONS: 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.

[Abnormal expression of RSK-4 and its clinical significance in breast cancer].

OBJECTIVE: To study the expression and clinical significance of ribosomal S6 kinase-4 (RSK-4) in breast cancer and explore the role of RSK-4 in the genesis and development of breast cancer. METHOD: The expression levels of RSK-4 mRNA and protein were detected in 56 cases of breast cancer and the normal breast tissues, as well as in 20 cases of breast benign lesions, by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. RESULTS: The expression rates of RSK-4 mRNA in breast cancer, the normal breast tissues and breast benign lesions were 48.2%, 76.8% and 75.0%, respectively. The expression level of RSK-4 mRNA in breast cancer was significantly lower than those in normal breast tissues and breast benign lesions tissues (P < 0.05). The expression level of RSK-4 significantly correlated with tumor size and clinical stage (P < 0.05).The expression rate of RSK-4 protein was 39.3% in breast cancer tissues, which was significantly lower than that of normal breast tissues (71.4%) and breast benign lesions (75.0%, P < 0.01). The expression level of RSK-4 protein was lower in breast cancer with large tumor, high clinical stage and lymph node metastasis. In 56 cases of breast cancer samples, the consistency rate of RSK-4 mRNA and protein was 73.2%. A significant correlation was found between RSK-4 mRNA and protein (χ² = 10.254, P < 0.05). CONCLUSION: The down-regulation of RSK-4 expression in breast caner suggests that it is a breast cancer suppressor gene, and the lack or down-regulation of RSK-4 expression is involved in the genesis and progression of breast cancer.

S100P as a novel biomarker of microvascular invasion and portal vein tumor thrombus in hepatocellular carcinoma.

BACKGROUND: Portal vein tumor thrombus (PVTT) and microvascular invasion (MVI) are types of intrahepatic vascular metastasis of hepatocellular carcinoma (HCC) and are highly correlated with poor prognosis. However, the underlying biomarkers of PVTT and MVI are unclear. METHODS: We identified a PVTT/MVI-associated gene S100P by cDNA microarray analysis, and assess the potential value of serum S100P measurement in the differential diagnosis of HCC and prediction of MVI status with large retrospective and perspective cohort studies. RESULTS: The mRNA and protein of S100P was increased in HCCs with PVTT or MVI. High S100P immunostaining in tumors was correlated with inferior tumor-free survival. Serum S100P values discriminated patients with HCCs from those with benign liver tumors, and it showed predictive potential of MVI status in both retrospective and perspective cohorts. S100P may regulate HCC tumorigenicity and invasive ability; S100P also was associated with up-regulation of CD44, which may mediate HCC cell adhesion to form PVTT/MVI. CONCLUSIONS: Serum S100P may be a novel differential diagnostic marker for HCC and a potential predictor of MVI status pre-surgery for HCC patients. S100P overexpression in HCC is highly correlated with the formation of PVTT and MVI, which may make S100P as a potential therapeutic target for HCC metastasis.

Outcomes of anatomical versus non-anatomical resection for hepatocellular carcinoma according to circulating tumour-cell status.

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.

A combination of subcuticular suture and enhanced recovery after surgery reduces wound complications in patients undergoing hepatectomy for hepatocellular carcinoma.

The aim of this study was to examine whether using subcuticular sutures during initial hepatectomy for hepatocellular carcinoma is associated with shorter postoperative length of hospital stay (PLOS) than using staples for patients treated in the enhanced recovery after surgery (ERAS) approach. A total of 376 patients were randomized to receive either subcuticular sutures or staples (188 per group), and the two groups were compared in terms of the incidence of wound complications and PLOS. Independent risk factors for PLOS were identified by multivariate analysis. Sutures were associated with significantly lower incidence of wound infection (4.3% vs. 13.3%, P = 0.020) and significantly shorter PLOS (7.97 vs. 8.45 days, P = 0.048). Independent risk factors for wound infection after hepatectomy were advanced age, increased preoperative body mass index, decreased preoperative serum albumin, and skin closure using staples. These results suggest that subcuticular sutures may be more effective than staples for conducting hepatectomy in patients with hepatocellular carcinoma who receive ERAS care.

Effect of KI-67 positive cellular index on prognosis after hepatectomy in Barcelona Clinic Liver Cancer stage A and B hepatocellular carcinoma with microvascular invasion.

OBJECTIVE: This 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). METHODS: A total of 333 patients who underwent curative hepatectomy had their immunohistochemistry analyzed retrospectively for KI-67 positive cellular index. RESULTS: In 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. CONCLUSION: Our 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.

Circulating Tumor Cells Undergoing EMT Provide a Metric for Diagnosis and Prognosis of Patients with Hepatocellular Carcinoma.

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 in situ 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.Significance: 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. Cancer Res; 78(16); 4731-44. ©2018 AACR.

Long-term survival and prognosis for primary clear cell carcinoma of the liver after hepatectomy.

BACKGROUND: The 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. METHODS: Our 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. RESULTS: The 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. CONCLUSION: Our 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.

Genetic polymorphisms -137 (rs187238) and -607 (rs1946518) in the interleukin-18 promoter may not be associated with development of hepatocellular carcinoma.

This study meta-analyzed the literature on possible association of polymorphisms -137 (rs187238) and -607 (rs1946518) in the interleukin-18 (IL-18) promoter with risk of hepatocellular carcinoma (HCC). The analysis included 8 case-control studies on the -137 polymorphism (1,318 cases, 2,254 controls) and 7 case-control studies on the -607 polymorphism (1,262 cases, 1,696 controls). None of the five genetic models suggested a significant association between the -137 polymorphism and HCC risk: allelic model, OR 0.99, 95% CI 0.74-1.34, P = 0.97; recessive model, OR 0.98, 95% CI 0.65-1.46, P = 0.91; dominant model, OR 1.35, 95% CI 0.73-2.52, P = 0.34; homozygous model, OR 0.99, 95% CI 0.65-1.49, P = 0.95; heterozygous model, OR 0.99, 95% CI 0.66-1.48, P = 0.94. Similar results were obtained in subgroup analyses of Asian patients, Chinese patients, or patients with hepatitis B virus (HBV)-related HCC. Similar results were also obtained for the -607 polymorphism across the entire study population as well as in the three subgroups. The available evidence suggests no significant association of the -137 or -607 polymorphisms with risk of HCC in general or specifically of HBV-related HCC. These conclusions should be verified in large, well-designed studies.

Primary hepatic non-Hodgkin's lymphoma with rectal cancer: A case report.

Primary hepatic non-Hodgkin's lymphoma (NHL) is an extremely rare disease that is commonly neglected as a possible diagnosis. The present study reports the case of a middle-aged male with chronic hepatitis B in which primary hepatic NHL and rectal cancer occurred simultaneously. A large solitary tumor in the left lobe of the liver was incidentally detected on routine examination prior to the laparoscopic resection of the rectal cancer. Laparoscopic resection of the rectal cancer and a liver biopsy were performed simultaneously. The pathology revealed that the hepatic tumor was NHL and that the rectal cancer was adenocarcinoma. Systemic staging revealed no evidence of nodal or bone marrow involvement, therefore, primary hepatic lymphoma (PHL) was diagnosed. PHL associated with rectal adenocarcinoma is extremely rare and to the best of our knowledge, has never been reported. At present, the cause and most effective therapy for the condition remain unclear.