Hepatocellular Carcinoma Group Research Articles

Current status of ultrasonography in national cancer surveillance program for hepatocellular carcinoma in South Korea: a large-scale multicenter study.

Abdominal ultrasonography (USG) is recommended as a surveillance test for high-risk groups for hepatocellular carcinoma (HCC). This study aimed to analyze the current status of the national cancer surveillance program for HCC in South Korea and investigate the effects of patient-, physician-, and machine-related factors on HCC detection sensitivity. This multicenter retrospective cohort study collected surveillance USG data from the high-risk group for HCC (liver cirrhosis or chronic hepatitis B or C >40 years of age) at eight South Korean tertiary hospitals in 2017. In 2017, 45 experienced hepatologists or radiologists performed 8,512 USG examinations. The physicians had a mean 15.0±8.3 years of experience; more hepatologists (61.4%) than radiologists (38.6%) participated. Each USG scan took a mean 12.2±3.4 minutes. The HCC detection rate by surveillance USG was 0.3% (n=23). Over 27 months of follow-up, an additional 135 patients (0.7%) developed new HCC. The patients were classified into three groups based on timing of HCC diagnosis since the 1st surveillance USG, and no significant intergroup difference in HCC characteristics was noted. HCC detection was significantly associated with patient-related factors, such as old age and advanced fibrosis, but not with physician- or machine-related factors. This is the first study of the current status of USG as a surveillance method for HCC at tertiary hospitals in South Korea. It is necessary to develop quality indicators and quality assessment procedures for USG to improve the detection rate of HCC.

The investigation on the hypercoagulability of hepatocellular carcinoma‐related cerebral infarction with thromboelastography

To investigate the hypercoagulability of hepatocellular carcinoma (HCC)-related cerebral infarction (HCRCI) with thromboelastography (TEG). A multicenter prospective study was conducted in HCRCI patients, HCC patients without cerebral infarction, and acute cerebral infarction (ACI) patients without HCC between January 2016 and December 2019. TEG parameters and laboratory and clinical data were collected and compared among the three groups. To confirm the independent risk factors of HCRCI, multivariate analyses were conducted. Receiver operating characteristic (ROC) curves were utilized to evaluate the area under the curve (AUC) plotted by each independent risk factor. There were 38 patientsrecruited in the HCRCI group, and 152 patients were recruited to the HCC group and the ACI group. The levels of plasma neutrophil count, D-dimer, α-fetoprotein (AFP), carcinoembryonic antigen, and maximum amplitude (MA)-a parameter of TEG-were significantly higher in the HCRCI group than HCC and ACI groups. Multivariate logistic regression analysis showed that increased neutrophile count, D-dimer, AFP, and MA were independently associated with HCRCI. ROC curve analysis showed firstthat AUC of MA for HCRCI was .875, which was larger than the other risk factors, and second that the optimal cutoff value for MA was 61.35, with a sensitivity of 89.50% and specificity of 66.40%. It was suggested that TEG disclosed that the pathogenesis of HCRIC is exactly related to the hypercoagulability. And with a cutoff value of MA equaling to 61.35, TEG facilitatesclinicians to identify HCC patients at high risk of HCRIC.

Role of textural analysis parameters derived from FDG PET/CT in differentiating hepatocellular carcinoma and hepatic metastases.

Texture and radiomic analysis characterize the tumor's phenotype and evaluate its microenvironment in quantitative terms. The aim of this study was to investigate the role of textural features of 18F-FDG PET/computed tomography (CT) images in differentiating hepatocellular carcinoma (HCC) and hepatic metastasis in patients with suspected liver tumors. This is a retrospective, single-center study of 30 patients who underwent FDG PET/CT for the characterization of liver lesions or for staging a suspected liver tumor. The histological diagnosis of either primary or metastatic tumor was obtained from CT-guided biopsy, ultrasound-guided biopsy, or surgical removal of a liver lesion. The PET/CT images were then processed in commercially available textural analysis software. Region of interest was drawn over the primary tumor with a 40% threshold and was processed further to derive 42 textural and radiomic parameters. These parameters were then compared between HCC group and hepatic metastases group. Receiver-operating characteristic (ROC) curves were used to identify cutoff values for textural features with a P value <0.05 for statistical significance. A retrospective study of 30 patients with suspected liver tumors was done. After undergoing PET/CT, the histological diagnosis of these lesions was confirmed. Among these 30 patients, 15 patients had HCC, and 15 patients had hepatic metastases from various primary sites. Seven textural analysis parameters were significant in differentiating HCC from liver metastasis. Cutoff values were calculated for these parameters according to the ROC curves, standardized uptake value (SUV) Skewness (0.705), SUV Kurtosis (3.65), SUV Excess Kurtosis (0.653), gray-level zone length matrix_long zone emphasis (349.2), gray-level zone length matrix_long zone low gray-level emphasis (1.6), gray-level run length matrix_long run emphasis (1.38) and gray-level co-occurrence matrix_Homogeneity (0.406). Textural analysis parameters could successfully differentiate HCC and hepatic metastasis non-invasively. Larger multi-center studies are needed for better clinical prognostication of these parameters.

A novel risk score model based on pyroptosis-related genes for predicting survival and immunogenic landscape in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is the third leading cause of cancer worldwide, with high incidence and mortality. Pyroptosis, a form of inflammatory-regulated cell death, is closely associated with oncogenesis. Expression profiles of HCC were downloaded from the TCGA database and validated using the ICGC and GEO databases. Consensus clustering analysis was used to determine distinct clusters. The pyroptosis-related genes (PRGs) included in the pyroptosis-related signature were selected by univariate Cox regression and LASSO regression analysis. Kaplan-Meier and receiver operating characteristic (ROC) analyses were performed to estimate the prognostic potential of the model. The characteristics of infiltration of immune cells between different groups of HCC were explored. Two independent clusters were identified according to PRG expression. Cluster 2 showed upregulated expression, poor prognosis, increased immune cell infiltration and worse immunotherapy response than cluster 1. A prognostic risk signature consisting of five genes (GSDME, NOD1, PLCG1, NLRP6 and NLRC4) was identified. In the high-risk score group, HCC patients showed decreased survival rates. In particular, multiple clinicopathological characteristics and immune cell infiltration were significantly associated with the risk score. Notably, the 5 PRGs in the risk score have been implicated in carcinogenesis, immunological pathways and drug sensitivity. A prognostic signature comprising five PRGs can be used as a potential prognostic factor for HCC. The PRG-related signature provides an in-depth understanding of the association between pyroptosis and chemotherapy or immunotherapy for HCC patients.

Evaluation of the safety and efficacy of ultrasound-guided percutaneous radiofrequency ablation for hepatocellular carcinoma and liver metastases adjacent to the gallbladder

Objective To evaluate the safety and efficacy of ultrasound-guided percutaneous radiofrequency ablation (RFA) in patients with hepatocellular carcinoma (HCC) and liver metastases adjacent to the gallbladder (GB). Materials and methods A total of 113 patients with 118 liver lesions (63 HCC lesions and 55 liver metastases) adjacent to the gallbladder underwent RFA between March 2011 and June 2019. Gallbladder-related complications and technique effectiveness rates were evaluated based on the classification of liver tumors and the distance between the lesion and the gallbladder. Results Gallbladder-related complications were observed in 13 patients. Among the patients with HCC, there was no significant difference between the ≤0.5 cm and >0.5 cm groups (p = .282). However, among the patients with liver metastases, the incidence of gallbladder-related complications in the ≤0.5 cm group was significantly higher than that in the >0.5 cm group (p = .025). The overall incidence of complications was significantly higher in the ≤0.5 cm group than in the >0.5 cm group (p = .020). Among the patients with lesions ≤3 cm, the technical effectiveness rate in the HCC group was significantly higher than in the liver metastasis group (p = .036). Conclusion RFA is a safe and effective treatment option for liver tumors adjacent to the gallbladder. Patients with lesions ≤0.5 cm from the gallbladder had higher gallbladder-related complications, especially patients with liver metastases. Among patients with lesions ≤3 cm, RFA showed greater technical effectiveness for treating HCC than for treating liver metastases.

Frizzled-3 suppression overcomes multidrug chemoresistance by Wnt/β-catenin signaling pathway inhibition in hepatocellular carcinoma cells

Multidrug resistance (MDR) is a major obstacle to the efficacy of hepatocellular carcinoma (HCC) chemotherapy. Previous studies have identified that low FZD3 predicted decreased survival after intraperitoneal versus intravenous-only chemotherapy in ovarian cancer. This study aimed to identify a potential target in HCC chemotherapy. The FZD3 expression variant in HCC cell lines was detected by RT-qPCR and western blotting. The FZD3 expression in the early recurrent HCC group (RE group) and the non-early recurrent HCC group (non-RE group) was measured by RT-qPCR. Then, the 50% inhibitory concentrations (IC50) in HCC cell lines were studied by MTT assay. TOP/FOP FLASH luciferase assay was performed to measure TCF-binding activities. We found that FZD3 was upregulated in three HCC cell lines, and the FZD3 expression was significantly higher in the RE group than in the non-RE group (P = 0.0344). A positive correlation between FZD3 and MDR1 was observed in HCC tissues (R2 = 0.6368, P = 0.0001). Then, we found that FZD3 knockdown significantly altered Huh-7 cell chemotherapeutic sensitivity to cisplatin [50.43 µM in the FZD3 siRNA (siFZD3) group vs 98.59 µM in the siRNA negative control (siNC) group; P = 0.007] or doxorubicin (7.43 µM in the siFZD3 group vs 14.93 µM in the siNC group; P = 0.017). TOP/FOP FLASH luciferase assay showed FZD3 could inhibit Wnt/β-catenin signaling in HCC cells. Moreover, FZD3 expression knockdown in SNU-449 and Huh-7 cells markedly reduced β-catenin and phosho-β-catenin (S37) protein expression, and Cyclin D1, c-myc and MDR1 were significantly decreased. This is the first study to describe the significantly increased FZD3 expression in patients with early recurrent HCC. FZD3 knockdown led to increased sensitivity to chemotherapy by Wnt/β-catenin signaling inhibition in HCC cell lines. Our study suggests FZD3 as a potential target for reversing chemoresistance in HCC.

Utility of comet assay of DNA damage in the detection of malignant transformation of chronic liver cirrhosis

Early detection of hepatocellular carcinoma (HCC) remains a great challenge in laboratory medicine. This study aimed to assess the ability to use the degree of DNA damage (using the comet assay) for the early detection of malignant transformation of liver cirrhosis (LC) to HCC. We used alkaline comet assay for measuring DNA damage in peripheral blood lymphocytes in 50 patients with chronic LC and 50 patients with HCC. Fifty healthy individuals served as a control group. We compared the results of comet assay parameters with alpha fetoprotein as a relevant traditional marker. The HCC group was associated with a significantly higher tail intensity (p=.004), tail moment (p=.016), total area (p=.003), total intensity (p=.010), width (p=.005), and a significantly lower head intensity (.004) when compared to the LC group. Good areas under the curve (AUCs) were found for total area (0.890), head intensity (0.880) and tail intensity (0.880), making it useful for discrimination between HCC and LC groups. Lower head intensity, higher tail intensity, tail moment, total area and width were found to be independent risk factors for HCC on top of LC. Measuring DNA damage using the Alkaline comet assay technique can be considered a sensitive and reliable diagnostic tool for early neoplastic transformation of advanced LC.

Characterization of Circulating Tumor Cells Using Imaging Flow Cytometry in Liver Disease Patients

Hepatocellular carcinoma (HCC) is asymptomatic at an early stage which delays its timely diagnosis and treatment. Circulating tumor cells (CTCs), derived from a primary or secondary tumor, may help in the management of HCC. Here, we evaluate and characterize CTCs in liver disease patients. In total, 65 patients, categorized into liver cirrhosis (LC) (n=30) and HCC (n=35), were enrolled. Using ImagestreamX MkII imaging flow cytometer, CTCs were detected and characterized using biomarker expression of EpCAM, CK, AFP, CD45, and DRAQ5 in LC and HCC patients. CTCs were detected in 33/35 (94%) HCC patients and in 28/30 (93%) LC patients. In the HCC group, the number of biomarker-positive CTCs was higher in BCLC stage D when compared with others. EpCAM+CK was the most expressed biomarker on CTCs in LC versus HCC (83.3% vs. 77.14%), followed by AFP (80% vs. 65.71%), EpCAM (30% vs. 28.57%), and CK (16.6% vs. 14.28%). The EpCAM cell area was significantly associated (P value=0.031) with the CTC-positive status. The combination biomarker expression of CTCs cell area (EpCAM, CK, and AFP) performed well with the area under the curve of 0.92, high sensitivity, and specificity in detecting early-stage and AFP-negative HCC as well as in AFP-negative LC cases. Enumeration and cell area of CTCs may be used as a biomarker for early detection of HCC and guiding treatment.

Transforming Growth Factor Β as a Marker of Hepatocellular Carcinoma in Patients with Chronic Hepatitis C Virus Infection.

The Transforming Growth Factor-beta (TGF-β) is one of the main growth factors associated with fibrosis or cirrhosis progression in the liver, but its role in hepatocarcinogenesis is controversial. To highlight the role of Transforming Growth Factor β as a marker of Hepatocellular carcinoma (HCC) in patients with chronic hepatitis C virus (HCV) infection. Ninety subjects were enrolled in this study, classified into three groups: Group I (chronic HCV group) included 30 patients with chronic HCV infection; Group II (HCC group) include 30 patients having HCC and chronic HCV infection and Group III consisted of 30 age and sex-matched healthy controls. TGF-β was evaluated in all the enrollees and its levels were correlated to liver function and other clinical parameters. TGF-β was found significantly higher in HCC group than in control and chronic HCV (P<0.001). In addition, it was correlated with biochemical and clinical parameters of cancer. Patients with HCC showed increased level of TGF-β compared to chronic HCV infection patients and controls.

Glypican-3, Vascular Endothelial Growth Factor and Golgi Protein-73 for Differentiation between Liver Cirrhosis and Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) accounts for more than 80% of primary liver cancers. Moreover, in the next 10 years, more than one million patients are expected to die from liver cancer as estimated by the World Health Organization. The aim of the present study is to evaluate the clinical utility of using Glypican (GPC3), Vascular endothelial growth factor (VEGF) and Golgi protein 73 (GP73) in serum by Enzyme-Linked Immunosorbent Assay (ELISA) and by Real-Time Polymerase Chain Reaction (RT-PCR), as diagnostic markers to differentiate HCC from cirrhotic liver disease. A total of 50 patients with histologically-proven HCC, 50 liver cirrhosis patients and 20 healthy volunteers as controls were enrolled in this study, blood samples were obtained from each patient. Expression of the studied biomarkers was evaluated by ELISA and Real-Time PCR. Statistical analysis of RT-PCR results showed that the expression of GPC3, VEGF and GP73 in serum of patients with HCC was significant (P value < 0.001, 0.01, and < 0.001) respectively and increased when compared to the cirrhotic group. Furthermore, the serum protein levels of GPC3 and VEGF in HCC and cirrhotic patients were significant when compared to the control group. While no significance was found between HCC and cirrhotic group. The serum protein level of GP73 was significantly increased in HCC and cirrhosis groups compared to the control group (P value < 0.001). Moreover, a significant increase was evident in HCC group compared to cirrhotic group (P value < 0.001). The results of the present study showed that the combination of VEGF and GP73 could discriminate HCC from cirrhosis. GPC3, VEGF and GP73 are reliable biomarkers for diagnosis of HCC. The serum level of GP73 is a potential screening marker for discriminating HCC from liver cirrhosis.

The Value of Thioredoxin Level and its Gene Polymorphism in the Diagnosis of Post- HCV Hepatocellular Carcinoma

Background: Hepatocellular carcinoma (HCC) is one of the most common malignancies and a leading cause of cancer-related death worldwide. Indeed, we need a novel tumor marker other than AFP for early detection and to improve the outcome. Serum thioredoxin is a promising protein involved in the pathogenesis of many malignancies. The study aims to evaluate serum thioredoxin and its gene polymorphism in HCC in cirrhotic patients due to HCV infection. Patients and Methods: 350 patients with HCC, 350 patients with chronic liver diseases, and 300 healthy controls were enrolled in our study. Serum thioredoxin level was measured by ELISA, and molecular study of thioredoxin domain-containing 5 (TXNDC5) gene polymorphism (rs1225943) polymorphism using real-time polymerase chain reaction by Taqman allele discrimination was done for all subjects. Results: Our study revealed a significant increase in serum thioredoxin levels in patients with HCC compared to chronic liver diseases and healthy controls. Using the Receiver operating characteristic (ROC) curve at the area under the curve (AUC) 0.917 and a cut-off value of &gt; 14.6 U/ml, our overall sensitivity and specificity for the HCC group over the other groups were 86 % and 92.15%, respectively with 92.2% positive predictive value and 54.9% negative predictive value. The molecular study of TXNDC5 gene polymorphism (rs1225943) polymorphism revealed no significant difference between the studied groups. Conclusion: Serum thioredoxin may be used as a promising tumor marker for HCC. Future research is needed to assess its use as a single or combined with other markers in the diagnosis and follow-up of the patients after interventions.

A Cross-Sectional Study on Association between Metabolic Syndrome and Hepatobiliary Cancers

Background and Aim: Several studies have supported the idea that a metabolic abnormality contributes to expansion of hepatocellular carcinoma (HCC). To date, metabolic factors have not been shown to be associated with hepatocellular carcinoma (HCC). Therefore, the present study aimed to assess the metabolic syndrome association with hepatocellular carcinoma. Patients and Methods: This cross-sectional study was carried out on surgically treated and radiologically confirmed 360 HCC cases in Gastroenterology Department of Central Hospital, Sargodha and Hayatabad Medical Complex Peshawar, during the period from January 2022 June 2022. Study protocol was approved by the research and ethical committee. Informed written consents were taken from each individual. Body mass index, blood pressure, medical history, and other related information were gathered from medical records. Patients were categorized into different groups based on their BMI: i) &lt;18.5 kg/m2, ii) 18.5-23.9 kg/m2, iii) 24-29.9 kg/m2, Fiv) 30-34.9 kg/m2 and ≥ 35 kg/m2. A routine biochemical or immunological analytic method was used to determine all metabolic parameters and liver function tests. The metabolic-associated factors were associated with HCC-related different liver function’s test and stratified for BMI, free fatty acids, and Glycated Albumin (GA). Chemical analysis was used to detect antioxidant capacity (TAOC) and malondialdehyde (MDA). Data analysis was done in SPSS version 27. Results: Of the total 360 HCC patients, there were 234 (65%) male and 126 (35%) females. The overall mean age was 68.62±6.4 years. The association between hepatobiliary cancer and body mass index (BMI), smoking, ischemic heart disease, hypertension, and diabetes was found in a univariate analysis of the data. Based on BMI, patients were categorized as follows: 35 (9.7%) in (&lt;18.5 kg/m2), 70 (19.4%) in (18.5-23.5 kg/m2), 118 (32.8%) in (24-29.9 kg/m2), 88 (24.4%) in (30-34.9 kg/m2), and 49 (13.7%) in (≥ 35 kg/m2). The incidence of smokers, non-smokers, and ex-smokers were 146 (40.6%), 60 (16.7%), and 154 (42.8%) respectively. The prevalence of different risk factors such as hypertension, diabetes, ischemic heart disease, Aspirin use, and statin use was 188 (52.2%), 124 (34.4%), 66 (18.3%), 148 (41.1%), and 142 (39.4%) respectively. The HCC group had significantly higher mean body mass index, glucose level, and lipid level than controls (P &lt;0.05). HCC patients showed a significant association between liver function and metabolic factors. A significant association between high level of GA and increased cancer risk were found between cases and control. Conclusion: The present study concluded that hepatobiliary carcinoma may be caused by metabolic syndrome, which includes dyslipidemia, a high body mass index, and high serum glucose levels as independent risk factors. This finding supports previous research and emphasizes that the connection between hapotabiliatry cancer and increased BMI remains for liver tumors, bile duct cancer, and gallbladder cancers. Keywords: Hepatobiliary carcinoma, metabolic syndrome, Obesity, Diabetes

Decreased granzyme-B expression in CD11c+CD8+ T cells associated with disease progression in patients with HBV-related hepatocellular carcinoma.

CD11c+CD8+ T cells are an unconventional CD8+ T cell subset that exerts antiviral activity in infectious diseases. However, its characteristics in hepatocellular carcinoma (HCC) have not been elucidated. Twenty-six patients with hepatitis B virus (HBV)-related HCC and 25 healthy controls (HC) were enrolled. The frequency and phenotype of CD11c+CD8+ T cells in peripheral blood and tumors in situ were detected by flow cytometry and immunohistochemistry. Both the HCC group and HC group had similar frequency and phenotype characteristics of CD11c+CD8+ T cells in the periphery. CD11c+CD8+ T cells were mainly composed of effector T cells, most of which were CD45RA+CCR7-. Compared with CD11c-CD8+ T cells, CD11c+CD8+ T cells had a higher proportion of CD38 and HLA-DR double positive, and expressed high levels of granzyme-B (GB) and degranulation marker CD107a, and produced high levels of interleukin-2 (IL-2), tumor necrosis factor alpha (TNF-α) and interferon-gamma (IFN-γ). However, the ability of degranulation and TNF-α production of CD11c+CD8+ T cells in patients with HCC were significantly lower than that in healthy controls. The GB expression level of peripheral CD11c+CD8+ T cells in patients with advanced stage of HCC was significantly lower than that in patients with early stage of HCC, and the GB expression level of liver-infiltrating CD11c+CD8+ T cells in tumor tissues was lower than that in non-tumor tissues. More importantly, the GB expression level of peripheral CD11c+CD8+ T cells was negatively correlated with tumor volume. These findings indicate that CD11c+CD8+ T cells may have potential anti-tumor activity and that GB+CD11c+CD8+ T cells are associated with disease progression in patients with HBV-related HCC.

Development and External Validation of a Radiomics Model Derived from Preoperative Gadoxetic Acid-Enhanced MRI for Predicting Histopathologic Grade of Hepatocellular Carcinoma.

Histopathologic grade of hepatocellular carcinoma (HCC) is an important predictor of early recurrence and poor prognosis after curative treatments. This study aims to develop a radiomics model based on preoperative gadoxetic acid-enhanced MRI for predicting HCC histopathologic grade and to validate its predictive performance in an independent external cohort. Clinical and imaging data of 403 consecutive HCC patients were retrospectively collected from two hospitals (265 and 138, respectively). Patients were categorized into poorly differentiated HCC and non-poorly differentiated HCC groups. A total of 851 radiomics features were extracted from the segmented tumor at the hepatobiliary phase images. Three classifiers, logistic regression (LR), support vector machine, and Adaboost were adopted for modeling. The areas under the curve of the three models were 0.70, 0.67, and 0.61, respectively, in the external test cohort. Alpha-fetoprotein (AFP) was the only significant clinicopathological variable associated with HCC grading (odds ratio: 2.75). When combining AFP, the LR+AFP model showed the best performance, with an AUC of 0.71 (95%CI: 0.59-0.82) in the external test cohort. A radiomics model based on gadoxetic acid-enhanced MRI was constructed in this study to discriminate HCC with different histopathologic grades. Its good performance indicates a promise in the preoperative prediction of HCC differentiation levels.

Impending Chemotherapeutic Impact of Arthrospira platensis Nanoparticles and/or Sorafenib against Hepatocellular Carcinoma through Modulation of Antioxidant Status, Tumor Marker Genes, and Anti-Inflammatory Signaling Pathways.

This study investigated Arthrospira platensis nanoparticles (NSP) to overcome sorafenib resistance in diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rats. This study used sixty Wistar male rats randomly grouped into two main groups, the normal control group, and the HCC model. For the normal control group (n = 12), animals were injected i.p. with PBS two times/week for 16 weeks. The remaining 48 rats were injected i.p. with using a single dose of diethyl nitrosamine (DENA) (200 mg/kg, ip), followed by phenobarbital sodium (0.05%) in drinking water for 16 weeks. At the end of the 16th week, rats were allocated into four groups (11 rats/each), one group was left without treatment (DENA group), and the other three groups were treated with either sorafenib (30 mg/kg; p.o.) or Arthrospira platensis Nanoparticles (NSP) (0.5 mg/kg body weight) once daily orally with the aid of gastric gavage or their combination for another four weeks. Blood and tissue samples were collected for further biochemical, histological, immunohistochemical, and gene expression analysis. Our result revealed that DENA-treated rats showed a marked elevation of hepatic enzyme markers with an increase in the total protein and globulin and decreases in the hepatic SOD. Catalase and GSH, with significantly increased MDA levels, subsequently increased the tumor biomarkers (AFP and CEA). On the molecular level, the DENA-treated rats showed significant up-regulation of Cyp19 mRNA and the inflammatory cytokines (TNF-α, iNOS, and TGF-1β) as well as the Ki-67 gene expression (p < 0.05) with down-regulation of the PPAR-γ and FOXO-1. In addition, the HCC group showed a loss of hepatic architecture, as well as atypia, swelling, macrosteatosis of hepatocytes, and fibrosis, besides increased vascularization. The immunohistochemical findings show increased expression of both GPC-3 and Hep Par 1 in the HCC group. SOR, NSP, or a combination of NSP and SOR.NSP treatment significantly overturned the DENA's harmful effect near the normal levels and restored all cancer biomarkers and antioxidant activities, indicating the chemotherapeutic impact of NSP. The present study provides evidence that NSP exerts a major anticancer effect on DENA-induced HCC. SOR/NSP is a promising combination for tumor suppression and overcoming sorafenib resistance in HCC by modulating antioxidants, anti-inflammatory signals, and tumor markers.

Incidental combined hepatocellular-cholangiocarcinoma in liver transplant patients: Does it have a worse prognosis?

Combined hepatocellular-cholangiocarcinoma (CHC) requires attention clinically and pathologically after liver transplantation (LT) because of its unique biology, difficulties in diagnosis, and being rare. We aimed to present our single-center experience for this incidental combined tumor. It is aimed to present our single-center experience for this incidental combined tumor. Seventeen patients with CHC were included in the study. There were 260 hepatocellular carcinoma (HCC) patients determined as the control group. Patients were evaluated for demographic, etiological, pathological features, and survival. Macrovascular and microvascular invasion levels were significantly higher in the CHC group (p<0.05). P53, CK19, and CK7 levels were significantly higher in the CHC group (p<0.05). Hepatocyte-specific antigen level was significantly higher in the HCC group. The mean overall survival was significantly higher in the HCC group (p<0.05). Even though CHC is a rare liver tumor, it has features that need to be clarified regarding both survival and tumor biology. İnvestigating prognostic factors, especially in terms of survival and recurrence, will be very beneficial to identify candidates who will benefit from LT and be included in the indications for LT for CHC. This study evaluated the outcomes of patients showing combined HCC-intrahepatic cholangiocarcinoma in explant pathology.

Feasibility of Raman spectroscopic identification of gall bladder cancer using extracellular vesicles extracted from bile.

Extracellular vesicles (EVs), which are heterogeneous membrane-based vesicles with bilayer cell membrane structures, could be versatile biomarkers for the identification of diverse diseases including cancers. With this potential, this study has attempted the Raman spectroscopic identification of gall bladder (GB) cancer by directly measuring the EV solution extracted from human bile without further sample drying. For this purpose, bile samples were obtained from four normal individuals and 21 GB polyp, eight hepatocellular carcinoma (HCC), and five GB cancer patients, and EVs were extracted from each of the bile samples. The Raman peak shapes of the EVs extracted from the GB cancer samples, especially the relative intensities of peaks in the 1560-1340 cm-1 range, were dissimilar to those of the samples from the normal, GB polyp, and HCC groups. The intensity ratios of peaks at 1537 and 1453 cm-1 and at 1395 and 1359 cm-1 of the GB cancer samples were lower and higher, respectively, than those of the samples of the remaining three groups. The differences of peak intensity ratios were statistically significant based on the Mann-Whitney U test. DNA/RNA bases, amino acids, and bile salts contributed to the spectra of EVs, and their relative abundances seemed to vary according to the occurrence of GB cancer. The varied metabolite compositions and/or structures of EVs were successfully demonstrated by the dissimilar peak intensity ratios in the Raman spectra, thereby enabling the discrimination of GB cancer.

Usefulness of Electron Density Calculated from Dual Energy CT in Differential Diagnosis between Hepatocellular Carcinoma and Hepatic Hemangioma

The aim of this study were to compare electron density (ED), obtained by dual energy computed tomography (DECT), between hepatocellular carcinoma (HCC) and hemangioma, and to assess the differential diagnostic performance of ED between HCC and hemangioma. A total of 46 patients (27 men and 19 women; mean age, 65.7±14.0 years) diagnosed with HCC or hemangioma who underwent upper abdominal DECT between October 2021 and December 2022 were included. ED of each lesion was measured. Relative ED (rED), which is normalized by the ED of background liver parenchyma, was calculated. ED and rED of HCC and hemangioma were statistically analyzed. The HCC group showed significantly higher ED (48.1±5.2) and rED (80.0±7.3) than the hemangioma group (43.7±4.1, 69.7±7.2, respectively) (p<0.01). The area under the curve of rED was greater than that of ED, but no significant difference was found (p=0.153). ED may help in the differential diagnosis between HCC and hemangioma.

Re-evaluation of the anticarcinogenic effect of metformin in a chemically-induced hepatocellular carcinoma model not associated with diabetes

ABSTRACT Hepatocellular carcinoma (HCC) is the fourth cause of cancer mortality worldwide. Metformin is an antidiabetic drug that showed a prophylactic effect in the early stages of some cancers. This study aimed to reevaluate the anticarcinogenic effect of metformin in a cancer model not associated with diabetes. Also, to evaluate the effect of pharmacological antidiabetic dose of metformin (used in cancer studies) at the early stages of HCC versus the sub-pharmacological dose. HCC was induced by a single i.p injection of diethylnitrosamine followed by 16 biweekly i.p injections of carbon-tetrachloride. Metformin was i.p injected (25 or 250 mg/kg) post HCC induction for four weeks. The low dose of metformin suppressed HCC by reducing liver biomarkers levels, and the number and size of hepatic nodules. Also, metformin (25 mg/kg) downstaged HCC via activating Bax and suppressing Bcl-2, VEGF, and PCNA. Mice treated with metformin (250 mg/kg) exhibited lactic acidosis, death, and insignificant differences in nodules’ number and size and immunohistochemical markers compared to the HCC group. This study concluded that the pharmacological dose of metformin cannot be used at advanced stages of HCC, while the sub-pharmacological dose of metformin showed a promising anticarcinogenic effect in chemically-induced HCC model not associated with diabetes. .

Clinical value of serum AFP and PIVKA‐II for diagnosis, treatment and prognosis of hepatocellular carcinoma

The accuracy of alpha-fetoprotein (AFP) as a diagnostic marker for hepatocellular carcinoma (HCC) is insufficient, and the application of abnormal prothrombin (PIVKA-II) in HCC is still controversial. Serum AFP and PIVKA-II levels were analyzed in 145 cases of HCC, 57 of benign liver disease, 55 of cholangiocarcinoma and gallbladder carcinoma, 112 of other gastrointestinal tumors with liver metastasis, and 101 healthy controls. Receiver operating characteristic curve and area under the curve were used to evaluate the diagnostic value of AFP and PIVKA-II for HCC. The changes in serum AFP and PIVKA-II levels before and after treatment in 47 HCC patients who underwent surgery and 77 who received interventional treatment were used to evaluate treatment efficacy and prognosis in HCC. The concentrations of AFP and PIVKA-II in the HCC group were significantly higher than those in other groups (p < 0.01). The diagnostic value of PIVKA-II in HCC was better than that of AFP, and combined detection improved the diagnostic sensitivity and specificity. The levels of AFP and PIVKA-II after liver cancer surgery were significantly lower than those before surgery. Elevated levels of PIVKA-II before surgery predicted disease progression, and patients who remained positive for PIVKA-II after surgery had worse prognosis than those who became negative after surgery. Combined detection of AFP and PIVKA-II is superior to both tests alone. We found that higher serum level of PIVKA-II indicates more severe HCC, with worse prognosis, while the level of AFP had no correlation with the prognosis.