Poorly-differentiated Hepatocellular Carcinoma Research Articles

The Combination of Gd-EOB-DTPA Enhanced T1 Mapping with Apparent Diffusion Coefficient could Improve the Diagnostic Efficacy of Hepatocellular Carcinoma Grading.

Accurately predicting the hepatocellular carcinoma (HCC) grade may facilitate the rational selection of treatment strategies. The diagnostic efficacy of the combination of Gadolinium ethoxybenzy diethylenetriamine pentaacetic (Gd-EOB-DTPA) enhancement T1 mapping and apparent diffusion coefficient (ADC) values in predicting HCC grade needs further validation. This study aimed to assess the capacity of Gd-EOB-DTPA-enhanced T1 mapping and ADC values, both individually and in combination, to discriminate between different grades of HCC. From July 2017 to February 2020, 96 patients (male, 83; mean age, 53.67 years; age range, 29-71 years) clinically diagnosed with HCC were included in the present study. All patients underwent Gd-EOB-DTPA-enhanced magnetic resonance imaging (MRI, including T1 mapping sequence) before surgery or biopsy. All the patients were categorized into 3 groups according to the pathological results (including 24 cases of well-differentiated HCCs, 59 cases of moderately differentiated HCCs, 13 cases of and poorly differentiated HCCs). The mean Gd-EOB-DTPA enhanced T1 values (&#916;T1=[(T1pre-T1post)/T1pre]×100%) and ADC values between different grading groups of HCC were calculated and compared. The area under the characteristics curve (AUC), the diagnostic threshold, sensitivity, and specificity of &#916;T1 and ADC for differential diagnosis were analyzed. Mean &#916;T1 was 58% for well-differentiated HCCs, 50% for moderately-differentiated HCCs, and 43% for poorly-differentiated HCCs. &#916;T1 showed statistical differences between the groups (P<0.001). The mean ADC values of the 3 groups were 1.11×10-3 mm2/s, 0.91×10-3 mm2/s, and 0.80×10-3mm2/s, respectively. ADC showed statistical differences between the groups (P<0.001). In discriminating well- differentiated group from the moderately differentiated group, the AUC of &#916;T1 was 0.751 (95% CI: 0.642, 0.859), the AUC of ADC was 0.782 (95% CI: 0.671, 0.894), the AUC of combined model was 0.811 (95% CI: 0.709, 0.914). In discriminating the poorly differentiated group from the moderately differentiated group, the AUC of &#916;T1 was 0.768 (95% CI: 0.634, 0.902), the AUC of ADC was 0.754 (95% CI: 0.603, 0.904), and the AUC of the combined model was 0.841 (95% CI: 0.729, 0.953). Gd-EOB-DTPA enhanced T1 mapping, and ADC values have complementary effects on the sensitivity and specificity for identifying different HCC grades. A combined model of Gd-EOB-DTPA-enhanced MRI T1 mapping and ADC values could improve diagnostic performance for predicting HCC grades.</p>.

Hepatocellular Carcinoma's Characteristics in an Endemic Country: A Closer Examination of Tumor Grade and Microvascular Invasion

Objective: Although histological grade and microvascular invasion are known predictors for patient survival and recurrence in hepatocellular carcinoma (HCC), their relationship with various clinical and histomorphological features of HCC remains unclear. Materials and Methods: Medical records were retrieved from 61 patients who were diagnosed with HCC from 2008-2018. Clinical and histomorphological variables that were hypothesized to be associated with histological grade and microvascular invasion were analyzed statistically using the Chi-square test or the Fisher’s exact test as alternatives. Multivariate analysis was performed with logistic regression model. Results: The majority of the patients had well to moderately-differentiated HCC (67.2%) with some of them presenting microvascular invasion (57.4%). Alpha-fetoprotein level (AFP) ≥100 ng/ml (p=0.036), tumor size &gt;7cm (p=0.031) and mitotic index ≥5 per 10 high power field (p=0.009) were significantly correlated with poorly-differentiated HCC. Mitotic Index ≥5 per 10 high power field was an independent factor for poorly differentiated HCC. Meanwhile BCLC stage B and mitotic index were also an independent factor for the presence of microvascular invasion. Conclusion: Larger tumor size and higher mitotic index was significantly correlated and independent factors for poorly differentiated HCC and microvascular invasion. In biopsy specimens for which the microvascular invasion is difficult to assess, histological grade, tumor size and mitotic index may be beneficial to depict the prognosis of patients with HCC.

Impact of portal-phase signal intensity of dynamic gadoxetic acid-enhanced magnetic resonance imaging in hepatocellular carcinoma.

To evaluate the prognostic impact of dynamic gadolinium ethoxybenzyl diethylenetriaminepentaacetic acid (Gd-EOB-DTPA)-enhanced magnetic resonance imaging (MRI) in patients with hepatocellular carcinoma (HCC). We retrospectively reviewed the data of 206 patients with HCC who underwent preoperative Gd-EOB-DTPA-enhanced MRI and hepatectomy and quantitatively evaluated the signal intensity ratio of the tumor to the surrounding liver tissue in the portal phase (SIRPP). We verified the survival rates and assessed the prognostic factors associated with overall survival (OS) and recurrence-free survival (RFS) using SIRPP. Multivariate analysis revealed that the independent predictive factors for poorly-differentiated HCC were α-fetoprotein > 20 ng/mL (hazard ratio [HR]: 3.1909, 95% confidence interval [CI]: 1.3464-7.5622, p = .0084) and SIRPP ≤ 0.85 (HR: 3.7155, 95% CI: 1.521-9.076, p = .004). The 5-year OS and RFS rates in the high and low SIRPP groups were 83.2 and 52.1%, respectively (p < .0001) and 49.7 and 18.5%, respectively (p = .0003). Multivariate analysis revealed that SIRPP ≤ 0.68 was an independent prognostic factor related to OS (HR: 4.4537, 95% CI: 1.6581-11.9626, p = .003). The SIRPP of preoperative Gd-EOB-DTPA-enhanced MRI might predict the histological differentiation and prognosis of HCC.

Nomogram based on Sonazoid contrast-enhanced ultrasound to differentiate intrahepatic cholangiocarcinoma and poorly differentiated hepatocellular carcinoma: a prospective multicenter study.

The aim of this study was to develop a predictive model based on Sonazoid contrast-enhanced ultrasound (SCEUS) and clinical features to discriminate poorly differentiated hepatocellular carcinoma (P-HCC) from intrahepatic cholangiocarcinoma (ICC). Forty-one ICC and forty-nine P-HCC patients were enrolled in this study. The CEUS LI-RADS category was assigned according to CEUS LI-RADS version 2017. Based on SCEUS and clinical features, a predicated model was established. Multivariate logistic regression analysis and LASSO logistic regression were used to identify the most valuable features, 400 times repeated 3-fold cross-validation was performed on the nomogram model and the model performance was determined by its discrimination, calibration, and clinical usefulness. Multivariate logistic regression and LASSO logistic regression indicated that age (>51 y), viral hepatitis (No), AFP level (≤ 20 µg/L), washout time (≤ 45 s), and enhancement level in the Kupffer phase (Defect) were valuable predictors related to ICC. The area under the receiver operating characteristic (AUC) of the nomogram was 0.930 (95% CI: 0.856-0.973), much higher than the subjective assessment by the sonographers and CEUS LI-RADS categories. The calibration curve showed that the predicted incidence was more consistent with the actual incidence of ICC, and 400 times repeated 3-fold cross-validation revealed good discrimination with a mean AUC of 0.851. Decision curve analysis showed that the nomogram could increase the net benefit for patients. The nomogram based on SCEUS and clinical features can effectively differentiate P-HCC from ICC.

Identification of novel phosphoproteomic biomarkers in patients with advanced hepatocellular carcinoma (HCC).

e15155 Background: HCC is one of the leading causes of cancer deaths worldwide. Despite increasing systemic therapy options for patients with advanced HCC, there are currently no predictive biomarkers to guide therapy selection. Phosphoproteomics has been successfully used to identify accurate predictive biomarkers in other cancers from frozen clinical samples (Dokal et al., 2021 ASCO Annual Meeting). In this study, we test the feasibility of biomarker discovery using phospho- and whole-proteomics analyses on formalin fixed and paraffin embedded (FFPE) tru-cut liver biopsies from patients with HCC. Methods: FFPE tru-cut biopsies (n = 29) were obtained from separate cohorts of 15 HCC patients [tumor (T)] and 14 patients with chronic liver disease with varying levels of cirrhosis, fibrosis or steatosis but without cancer [non-tumor (NT)]. T samples were from advanced HCC patients with varied aetiology (chronic viral hepatitis, alcohol or NAFLD). Proteins were extracted from 10 x 10µm sections; crosslinks reversed before digestion to peptides; and multiple clean-up/enrichment steps before analysis. Peptides were quantified by mass spectrometry and performance assessed using multi-variate and enrichment analyses. Results: Overall, 4978 phosphopeptides and 3721 proteins were quantified across all samples. Multi-variate analyses based on the relative expression of phosphopeptides or proteins showed a clear separation between T and NT samples, as well as phenotype-distinct separation (i.e according to differentiation) within the T group. Subsequent testing also verified that the method could reproducibly measure these HCC-differentiating phosphorylation sites (and proteins). Proteins associated with HCC were regulated as expected in T vs NT, e.g. ASS1 (log2-fold change (FC) = -1.75, p &lt; 0.001), BDH2 (log2-FC = -1.97, p &lt; 0.001). Known drug targets were found to be significantly upregulated in poorly-differentiated HCC, e.g. PARP1 (log2-FC = 1.7, p &lt; 0.001) and AKT1 (log2-FC = 1.5, p &lt; 0.001), as well as previously described phosphorylation sites, e.g. p-CDK1 T14 (log2-FC = 5.95, p = 0.007) and p-MCM2 S139 (log2-FC = 2.98, p = 0.046). Kinase substrate enrichment analysis (Casado et al., Science Signaling 2013) showed increased activity for several kinases in T vs NT, including A-RAF (z-score = 3.2, p &lt; 0.001), MAPK3 (z-score = 2.1, p = 0.022) and TNK2 (z-score = 2.1, p = 0.003). Conclusions: We successfully adapted (phospho-)proteomics to FFPE tru-cut biopsy specimens, enabling reproducible identification of novel and known (phospho-)proteins that distinguished differentiation status. A prospective study utilising this phosphoproteomic analysis pipeline is now underway, recruiting patients with advanced HCC receiving first line standard of care therapy with immune checkpoint inhibitor combinations or tyrosine kinase inhibitors to identify potential predictive biomarkers of response.

Pre-operative MRI features predict early post-operative recurrence of hepatocellular carcinoma with different degrees of pathological differentiation

PurposeTo investigate the value of pre-operative gadoxetate disodium (Gd-EOB-DTPA) enhanced MRI predicting early post-operative recurrence (< 2 years) of hepatocellular carcinoma (HCC) with different degrees of pathological differentiation.MethodsRetrospective analysis of pre-operative MR imaging features of 177 patients diagnosed as suffering from HCC and that underwent radical resection. Multivariate logistic regression assessment was adopted to assess predictors for HCC recurrence with different degrees of pathological differentiation. The area under the curve (AUC) of receiver operating characteristics (ROC) was utilized to assess the diagnostic efficacy of the predictors.ResultsAmong the 177 patients, 155 (87.5%) were males, 22 (12.5%) were females; the mean age was 49.97 ± 10.71 years. Among the predictors of early post-operative recurrence of highly-differentiated HCC were an unsmooth tumor margin and an incomplete/without tumor capsule (p = 0.037 and 0.033, respectively) whereas those of early post-operative recurrence of moderately-differentiated HCC were incomplete/without tumor capsule, peritumoral enhancement along with peritumoral hypointensity (p = 0.006, 0.046 and 0.004, respectively). The predictors of early post-operative recurrence of poorly-differentiated HCC were peritumoral enhancement, peritumoral hypointensity, and tumor thrombosis (p = 0.033, 0.006 and 0.021, respectively). The AUCs of the multi-predictor diagnosis of early post-operative recurrence of highly-, moderately-, and poorly-differentiated HCC were 0.841, 0.873, and 0.875, respectively. The AUCs of the multi-predictor diagnosis were each higher than for those predicted separately.ConclusionsThe imaging parameters for predicting early post-operative recurrence of HCC with different degrees of pathological differentiation were different and combining these predictors can improve the diagnostic efficacy of early post-operative HCC recurrence.

Boric acid exert anti-cancer effect in poorly differentiated hepatocellular carcinoma cells via inhibition of AKT signaling pathway

BackgroundThe possible anti-cancer properties of boron, a trace element for humans, have been demonstrated in various experimental and epidemiological studies, although the effects of boron on liver cancer are unclear. In the present study we evaluate the effects of boric acid on the cell lines of hepatocellular carcinoma (HCC) of the liver, as the leading form of liver cancer, for which a poorly-differentiated HCC cell line (Mahlavu cell line) was used. MethodsThe anti-cancer effect of boric acid was investigated with a cell viability assay, apoptosis analysis, cell migration analysis, cell morphology analysis, colony formation assay and 3D cell culture techniques. Also, the effect of boric acid on the AKT signaling pathway was determined through a western blot analysis. ResultsBoric acid was found to reduce cell viability in a dose- and time-dependent manner, and decreased survival, colony formation ability, migration capability and HCC cell tumor spheroid growth in HCC cell lines, while also inducing apoptosis, autophagy and morphological alteration. Furthermore, boric acid inhibited AKT phosphorylation, and anticancer biological responses in HCC cells were observed only in cells in which AKT phosphorylation was suppressed by boric acid. ConclusionOur results suggest that boric acid might be a promising therapeutic candidate in hepatocellular carcinoma via the inhibition of AKT signaling pathway.

Electrical conductivity spectra of hepatic tumors reflect hepatocellular carcinoma progression in mice

Background: Electrical impedance spectroscopy is a technique which evaluates differences in dielectric properties of tissues for cancer identification. Methods: Murine hepatic cancer model was developed by intraperitoneal administration of N-nitrosodiethylamine to male BALB/c mice. Tumors obtained were evaluated for their conductivity in frequency range of (4 Hz–5 MHz). All tumors were subjected to histopathological grading and parameters such as free spacing, necrosis, and cell density were estimated on histological slides. The status of gap junctions and gap junction intercellular communication (GJIC) were studied using enzyme-linked immunosorbent assay, immunohistochemistry, dye transfer assay, and electron microscopy. Results: Histopathological investigation revealed the presence of moderately to poorly-differentiated hepatocellular carcinoma (HCC) in mice. All types of tumors showed higher electrical conductivity than normal liver tissue in frequency range (4 Hz–1 kHz). However, in frequency range (10 kHz–5 MHz) only poorly-differentiated tumors showed higher conductivity compared to normal tissue. The most prominent findings in moderately-differentiated and poorly-differentiated HCC were increased visible free spaces and necrosis respectively. The status of cell gap junctions were significantly deteriorated in tumors and a corresponding significant reduction in GJIC was also observed. These biological indicators were correlated with electrical conductivity of hepatic tumors. Conclusion: Variations in electrical conductivity spectra of hepatic tumors reflect progression of HCC. General significance: Future studies can be planned to perform hierarchical clustering of dielectric parameters with more number of tumor samples to establish dielectric spectroscopy-based classification or staging of hepatic tumors.

Texture analysis of MR images to identify the differentiated degree in hepatocellular carcinoma: a retrospective study

BackgroundTo explore the clinical value of texture analysis of MR images (multiphase Gd-EOB-DTPA-enhanced MRI and T2 weighted imaging (T2WI) to identify the differentiated degree of hepatocellular carcinoma (HCC).MethodOne hundred four participants were enrolled in this retrospective study. Each participant performed preoperative Gd-EOB-DTPA-enhanced MR scanning. Texture features were analyzed by MaZda, and B11 program was used for data analysis and classification. The diagnosis efficiencies of texture features and conventional imaging features in identifying the differentiated degree of HCC were assessed by receiver operating characteristic analysis. The relationship between texture features and differentiated degree of HCC was evaluated by Spearman’s correlation coefficient.ResultsThe grey-level co-occurrence matrix -based texture features were most frequently extracted and the nonlinear discriminant analysis was excellent with the misclassification rate ranging from 3.33 to 14.93%. The area under the curve (AUC) of the combined texture features between poorly- and well-differentiated HCC, poorly- and moderately-differentiated HCC, moderately- and well-differentiated HCC was 0.812, 0.879 and 0.808 respectively, while the AUC of tumor size was 0.649, 0.660 and 0.517 respectively. The tumor size was significantly different between poorly- and moderately-HCC (p = 0.014). The COMBINE AUC values were not increased with tumor size combined.ConclusionsTexture analysis of Gd-EOB-DTPA-enhanced MRI and T2WI was valuable and might be a promising method in identifying the differentiated degree of HCC. The poorly-differentiated HCC was more heterogeneous than well- and moderately-differentiated HCC.

Dynamic Contrast-Enhanced Computed Tomography Findings that may Predict Poorly-Differentiated Hepatocellular Carcinoma Prior to Treatment

Objective: To identify CT findings that predict poorly-differentiated hepatocellular carcinoma (p-HCC).Methods: This retrospective study included pathologically proven HCC patients during January 2010 to December 2017 who underwent dynamic contrast-enhanced computed tomography (CT) imaging within 12 weeks before the pathological diagnosis. CT findings were reviewed and graded by consensus opinion of two abdominal radiologists. The relationship between imaging findings and histological differentiation of HCC was analyzed using chi-square test. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy for diagnosis of p-HCC were calculated.Results: Of 200 HCCs during the study period, 18 were well-differentiated, 170 were moderately-differentiated, and 12 were poorly differentiated. Irregular rim enhancement in arterial phase (p&lt;0.001) and presence of lymphadenopathy (p=0.003) were both statistically significantly different among the three types of histological differentiation of HCC. Sensitivity, specificity, PPV, NPV, and accuracy for prediction of p-HCC by the presence of irregular rim enhancement in arterial phase and lymphadenopathy were 58.3%, 97.3%, 58.3%, 97.3%, and 95%, and 50%, 88.8%, 22.2%, 96.5%, and 86.5% - all respectively.Conclusion: The presence of irregular rim enhancement in arterial phase and lymphadenopathy are potentially useful CT findings for prediction of p-HCC prior to treatment.

Correlation between p53 expression and&#x0D; hepatocellular carcinoma differentiation grade

Relevance: Hepatocellular carcinoma (HCC) is one of the world’s most common liver cancers and fatal malignant neoplasms. According to GLOBOCAN 2018, liver cancer ranks sixth in incidence (842,000 new cases) and fourth in mortality&#x0D; (782,000 deaths) globally. In Kazakhstan, there is an increase in HCC incidence from 879 cases in 2016 up to 984 cases in 2017,&#x0D; which amounted to 5.5 cases per 100 000 population. In 2017, the five-year survival rate was 23.7%.&#x0D; The mutations in the p53 gene that encodes the p53 protein are most frequent in HCC (35.2%). A high level of nuclear expression of p53 and a decrease in apoptosis of tumor cells with a simultaneous increase in their proliferative activity, noted in&#x0D; HCC cells, are important for the prognosis of the disease. There is a correlation between poor prognosis, a decreased survival of&#x0D; patients with HCC, and high expression of the p53 oncoprotein in HCC.&#x0D; U. Nzeako et al. have revealed a strong correlation of HCC histological differentiation with the patient survival rate. Therefore, a study of the correlation between p53 expression and HCC differentiation grade is very relevant.&#x0D; The purpose of this study was to analyze the correlation between p53 expression and HCC differentiation grade.&#x0D; Results: p53 was overexpressed in 18% of HCC cases; the expression was high in 62% and low in 20% of HCC cases. In&#x0D; well-differentiated HCC (G1), the p53-immunopositive nuclei expression was low in 10 cases and high in 2 cases. In moderately&#x0D; differentiated HCC (G2), p53 was highly expressed in 21 cases and overexpressed in 2 cases. In poorly-differentiated HCC (G3),&#x0D; p53 was overexpressed in 7 cases and highly expressed in 8 cases of HCC. The analysis of the correlation between p53 expression and HCC differentiation grade by Spearman showed a high correlation (r=0.79, p&lt;0.01), evidencing a direct dependence&#x0D; of p53 expression level on HCC gradation.&#x0D; Conclusion: HCC differentiation grade highly correlates with p53 expression level in immunopositive nuclei of HCC cells.&#x0D; The obtained data evidences a dependence between p53 expression level and HCC gradation.

Diagnostic value of glypican-3, arginase-1 and hepatocyte paraffin antigen -1 in differentiating hepatocellular carcinoma from intrahepatic cholangiocarcinoma.

BackgroundThe aim of the present study was to investigate the diagnostic value of glypican-3 (GPC3), arginase-1 (Arg-1), and hepatocyte paraffin antigen 1 (HepPar-1) in differentiating hepatocellular carcinoma (HCC) from intrahepatic cholangiocarcinoma (ICC).MethodsThe expression of GPC3, HepPar-1 and Arg-1 were measured by immunohistochemistry in 47 cases of HCC, 29 cases of ICC and their paracancerous tissues.ResultsA high expression of GPC3, Arg-1 and HepPar-1 was observed in HCC tissues (68.09%, 76.60% and 78.72%, respectively; P>0.0125) while it was lower in ICC tissues (6.90%, 6.90% and 13.79%, respectively; P>0.0125). With regard to specificity, GPC3 performed better than Arg-1 and HepPar-1 (97.37% vs. 1.32% and 2.63%, respectively; P<0.05). The positive rate in poorly differentiated HCC for either GPC3, Arg-1 or HepPar-1 was lower than that in well- and moderately differentiated HCC. The majority of positive samples for GPC3 and Arg-1 were grade 2+ in well-, moderately- or poorly-differentiated HCC. Combined detection of GPC3, Arg-1 and HepPar-1 could increase the sensitivity up to 89.36% and the specificity to 100.00%, comparing with any single biomarker (P<0.05).ConclusionsGPC3, Arg-1 and HepPar-1 were all useful biomarkers in differentiating HCC from ICC. The combination models could improve the diagnosis value of HCC and help differentiating HCC from ICC.

Analysis of competing endogenous RNA network identifies a poorly differentiated cancer-specific RNA signature for hepatocellular carcinoma.

Plenty of evidence has suggested that long noncoding RNAs (lncRNAs) play a vital role in competing endogenous RNA (ceRNA) networks. Poorly differentiated hepatocellular carcinoma (PDHCC) is a malignant phenotype. This paper aimed to explore the effect and the underlying regulatory mechanism of lncRNAs on PDHCC as a kind of ceRNA. Additionally, prognosis prediction was assessed. A total of 943 messenger RNAs (mRNAs), 86 miRNAs, and 468 lncRNAs that were differentially expressed between 137 PDHCCs and 235 well-differentiated HCCs were identified. Thereafter, a ceRNA network related to the dysregulated lncRNAs was established according to bioinformatic analysis and included 29 lncRNAs, 9 miRNAs, and 96 mRNAs. RNA-related overall survival (OS) curves were determined using the Kaplan-Meier method. The lncRNA ARHGEF7-AS2 was markedly correlated with OS in HCC (P = .041). Moreover, Cox regression analysis revealed that patients with low ARHGEF7-AS2 expression were associated with notably shorter survival time (P = .038). In addition, the area under the curve values of the lncRNA signature for 1-, 3-, and 5-year survival were 0.806, 0.741, and 0.701, respectively. Furthermore, a lncRNA nomogram was established, and the C-index of the internal validation was 0.717. In vitro experiments were performed to demonstrate that silencing ARHGEF7-AS2 expression significantly promoted HCC cell proliferation and migration. Taken together, our findings shed more light on the ceRNA network related to lncRNAs in PDHCC, and ARHGEF7-AS2 may be used as an independent biomarker to predict the prognosis of HCC.

Noninvasive Evaluation of the Pathologic Grade of Hepatocellular Carcinoma Using MCF-3DCNN: A Pilot Study.

Purpose To evaluate the diagnostic performance of deep learning with a multichannel fusion three-dimensional convolutional neural network (MCF-3DCNN) in the differentiation of the pathologic grades of hepatocellular carcinoma (HCC) based on dynamic contrast-enhanced magnetic resonance images (DCE-MR images). Methods and Materials Fifty-one histologically proven HCCs from 42 consecutive patients from January 2015 to September 2017 were included in this retrospective study. Pathologic examinations revealed nine well-differentiated (WD), 35 moderately differentiated (MD), and seven poorly differentiated (PD) HCCs. DCE-MR images with five phases were collected using a 3.0 Tesla MR scanner. The 4D-tensor representation was employed to organize the collected data in one temporal and three spatial dimensions by referring to the phases and 3D scanning slices of the DCE-MR images. A deep learning diagnosis model with MCF-3DCNN was proposed, and the structure of MCF-3DCNN was determined to approximate clinical diagnosis experience by taking into account the significance of the spatial and temporal information from DCE-MR images. Then, MCF-3DCNN was trained based on well-labeled samples of HCC lesions from real patient cases by experienced radiologists. The accuracy when differentiating the pathologic grades of HCC was calculated, and the performance of MCF-3DCNN in lesion diagnosis was assessed. Additionally, the areas under the receiver operating characteristic curves (AUC) for distinguishing WD, MD, and PD HCCs were calculated. Results MCF-3DCNN achieved an average accuracy of 0.7396±0.0104 with regard to totally differentiating the pathologic grade of HCC. MCF-3DCNN also achieved the highest diagnostic performance for discriminating WD HCCs from others, with an average AUC, accuracy, sensitivity, and specificity of 0.96, 91.00%, 96.88%, and 89.62%, respectively. Conclusions This study indicates that MCF-3DCNN can be a promising technology for evaluating the pathologic grade of HCC based on DCE-MR images.

18F-FDG PET/CT AS AN ASSESSMENT TOOL OF HEPATOCELLULAR CARCINOMA SECONDARY TO NON-ALCOHOLIC FATTY LIVER DISEASE DEVELOPMENT IN EXPERIMENTAL MODEL.

Hepatocellular carcinoma (HCC) can be the last step of non-alcoholic fatty liver disease (NAFLD) evolution. Experimental models are crucial to elucidate the pathogenesis of HCC secondary to NAFLD. The 2-deoxy-2-(18F)fluoro-D-glucose (18F-FDG) positron emission tomography/computed tomography (PET/CT) plays an important role in evaluating HCC development and progression. To standardize the imaging method of PET/CT with 18F-FDG as an evaluation tool of the experimental model of HCC secondary to NAFLD. Ten male Sprague-Dawley rats were fed with choline-deficient high-fat diet and diethylnitrosamine (DEN) in the drinking water for 16 weeks and then received 1 mL of saline solution (0.9%) daily by gavage for three weeks. At the 16th and 19th weeks, abdominal ultrasonography (USG) was performed. 18F-FDG PET/CT images were obtained before the beginning of experiment (week 0) and at the end (week 19). Histological and immunohistochemically analysis were also performed. The USG results showed a homogeneous group at the 16th week with an average of 4.6±2.74 nodules per animal. At the 19th week, PET/CT findings demonstrated an average of 8.5±3.7 nodules per animal. The mean values of SUVmed and SUVmax were 2.186±0.1698 and 3.8±1.74, respectively. The average number of nodules per animal in the histological analysis was 5.5±1.5. From all nodules, 4.6% were classified as well-differentiated HCC and 81.8% were classified as poorly-differentiated HCC. 18F-FDG PET/CT was able to evaluate the development of HCC in an experimental model of NAFLD non-invasively. From the standardization of PET/CT in this model, it is possible to use this tool in future studies to monitor, in vivo and non-invasively, the progression of HCC.

Indian Hedgehog links obesity to development of hepatocellular carcinoma.

Obesity increases the risk of hepatocellular carcinoma (HCC), but precise identification and characterization of druggable oncogenic pathways that contribute to the progression of NAFLD to HCC, and hence to the increased incidence and aggressiveness of HCC in obese individuals is lacking. In this regard, we demonstrate that the Indian Hedgehog (Ihh) signaling pathway is upregulated in the fatty livers of mice consuming a high fat diet, and furthermore sustained in HCC tumors specifically within the context of a NAFLD microenvironment. Using a diet-induced mouse model of HCC wherein only obese mice develop HCC, targeted ablation of hepatocyte-secreted Ihh results in a decreased tumor burden and lower grade tumors. Ihh activation regulates the transdifferentiation of ciliated stellate cells and proliferation of Epcam+ ductal cells to promote fibrosis. Mechanistically, increased expression of hitherto uncharacterized effectors of Hh pathway, namely Myc and Tgf-β2 is critical to the observed physiology. This pro-tumorigenic response is driven by increased expression of Wnt5a to effect a poorly-differentiated and invasive tumor phenotype. Wnt5a secreted from activated stellate cells act on Ror2-expressing hepatocytes. We further demonstrate that Wnt5a expression is also elevated in poorly-differentiated HCC cells, suggesting that these ligands are also able to function in an autocrine positive feedback manner to sustain poorly-differentiated tumors. Taken together, our study provides a mechanistic understanding for how Ihh signaling promotes HCC tumorigenesis specifically in obese mice. We propose that therapeutic targeting of the Hh pathway offers benefit for patients with dietary / NAFLD-driven steatotic HCC.

Liver cancer cell lines distinctly mimic the metabolic gene expression pattern of the corresponding human tumours

BackgroundAlthough metabolism is profoundly altered in human liver cancer, the extent to which experimental models, e.g. cell lines, mimic those alterations is unresolved. Here, we aimed to determine the resemblance of hepatocellular carcinoma (HCC) cell lines to human liver tumours, specifically in the expression of deregulated metabolic targets in clinical tissue samples.MethodsWe compared the overall gene expression profile of poorly-differentiated (HLE, HLF, SNU-449) to well-differentiated (HUH7, HEPG2, HEP3B) HCC cell lines in three publicly available microarray datasets. Three thousand and eighty-five differentially expressed genes in ≥2 datasets (P < 0.05) were used for pathway enrichment and gene ontology (GO) analyses. Further, we compared the topmost gene expression, pathways, and GO from poorly differentiated cell lines to the pattern from four human HCC datasets (623 tumour tissues). In well- versus poorly differentiated cell lines, and in representative models HLE and HUH7 cells, we specifically assessed the expression pattern of 634 consistently deregulated metabolic genes in human HCC. These data were complemented by quantitative PCR, proteomics, metabolomics and assessment of response to thirteen metabolism-targeting compounds in HLE versus HUH7 cells.ResultsWe found that poorly-differentiated HCC cells display upregulated MAPK/RAS/NFkB signaling, focal adhesion, and downregulated complement/coagulation cascade, PPAR-signaling, among pathway alterations seen in clinical tumour datasets. In HLE cells, 148 downregulated metabolic genes in liver tumours also showed low gene/protein expression – notably in fatty acid β-oxidation (e.g. ACAA1/2, ACADSB, HADH), urea cycle (e.g. CPS1, ARG1, ASL), molecule transport (e.g. SLC2A2, SLC7A1, SLC25A15/20), and amino acid metabolism (e.g. PHGDH, PSAT1, GOT1, GLUD1). In contrast, HUH7 cells showed a higher expression of 98 metabolic targets upregulated in tumours (e.g. HK2, PKM, PSPH, GLUL, ASNS, and fatty acid synthesis enzymes ACLY, FASN). Metabolomics revealed that the genomic portrait of HLE cells co-exist with profound reliance on glutamine to fuel tricarboxylic acid cycle, whereas HUH7 cells use both glucose and glutamine. Targeting glutamine pathway selectively suppressed the proliferation of HLE cells.ConclusionsWe report a yet unappreciated distinct expression pattern of clinically-relevant metabolic genes in HCC cell lines, which could enable the identification and therapeutic targeting of metabolic vulnerabilities at various liver cancer stages.

Hepatocellular Carcinoma: Retrospective Evaluation of the Correlation Between Gadobenate Dimeglumine-Enhanced Magnetic Resonance Imaging and Pathologic Grade.

The aim of this study was to evaluate the usefulness of gadobenate dimeglumine-enhanced magnetic resonance imaging in characterizing the grade of hepatocellular carcinoma (HCC) using the signal intensity (SI) of the erector spinae as internal reference. Clinical data of 40 patients (a total of 44 lesions) confirmed by pathology for HCC were retrospectively reviewed. Gadobenate dimeglumine-enhanced magnetic resonance imaging was performed in all patients, and SI of lesions (SIles), liver parenchyma around the lesions (SIhep), erector spinae (SImus) and standard deviation of SI of the surrounding noise (SDnoi) on nonenhanced T2WI, nonenhanced T1WI, and contrast-enhanced T1WI (in both arterial and hepatobiliary phase [AP and HBP]) were measured, respectively. Contrast-to-noise ratio (CNR) were separately defined as CNR1 ([SIles - SIhep]/SDnoi) and CNR2 ([SIles - SImus]/SDnoi). Statistical analyses were performed using one-way analysis of variance, least significant difference test, logistic regression analysis, Spearman rank correlation, and receiver operating characteristic curves analysis. Forty-four HCCs, including 3 well-differentiated HCCs, 26 moderately differentiated HCCs, and 15 poorly differentiated (PD) HCCs, were confirmed. On logistic regression analysis, only CNR2 in the HBP was predictor of PD HCCs (P = 0.015, odds ratio = 1.040). The size of lesions, CNR1 in the AP, CNR2 in the AP, and CNR2 in the HBP, showed significant correlations with the degree of differentiation (correlation coefficients = -0.371, 0.435, 0.503, and 0.512, respectively; P = 0.013, 0.003, 0.001, and 0.000, respectively). Contrast-to-noise ratio 2 in the HBP with the cutoff of less than 4.56 could distinguish moderately differentiated HCCs from PD HCC with the sensitivity and specificity of 84.6% and 60.0%, respectively. Relatively low arterial enhancement and low CNR2 value in the HBP are predictive for poor histological grade of HCCs.

Immunohistochemical Assessment of the Expression of Biliary Transportation Proteins MRP2 and MRP3 in Hepatocellular Carcinoma and in Cholangiocarcinoma

Multidrug resistance-associated protein 2 (MRP2) is a multi-specific organic anion transporter predominantly expressed in the canalicular membrane of hepatocytes, epithelial cells from gallbladder and apical membranes of proximal tubular kidney epithelium whereas multidrug resistance-associated protein 3 (MRP3) is present in the basolateral membrane of hepatocytes and cholangiocytes. This study aims to detect the expression of these transporters in hepatocellular carcinoma (HCC) and in cholangiocarcinoma (CC), searching for evidences for future studies on differential diagnosis and on clinical essays. The immunohistochemical reactivity (IHC) of these transporters was assessed in tissue microarrays of 80 HCC and 56 CC cases using monoclonal antibodies and compared with anatomopathological (AP) variables. The positivity of MRP2 was observed in 92.3% of HCC and in 96.3% of CC. The detection of high MRP2 expression in HCC was not significantly different (p > 0.05) according to the size, number of nodules architectural pattern and growth pattern of HCC and CC. Regarding histological grades, 22/22 well moderately differentiated HCC versus 50/56 poorly differentiated HCC were positive for MRP2. A trend for lower expression in poor differentiation HCC was found. And 50/50 well/moderately differentiated CC versus 2/4 poorly/undifferentiated CC were positive for MRP2. This result showed a reduced expression (p = 0,0004) in poorly differentiated CC. MRP3 positivity was observed in 18.8% of HCC and was not significantly different according to AP parameters. MRP3 was expressed in 44.5% CC, with a trend for lower expression in less differentiated CC and significantly lower rates in the ductular histological subtype (p = 0.023). The high expression of MRP2 in HCC and in CC is conserved regardless most of the anatomopathological parameters, except for a trend of lower expression in less differentiated HCC and CC. The observation of lower MRP3 expression in less differentiated CC and, especially, in the histological subtype with expression of hepatic progenitor cell phenotypes leads to future opportunities to evaluate the expression of this marker in cholangiocarcinomas.

Prediction of histological grade of hepatocellular carcinoma using quantitative diffusion-weighted MRI: a retrospective multivendor study.

To evaluate the usefulness of quantitative diffusion-weighted (DW) imaging acquired by multivendor magnetic resonance units for predicting grade of hepatocellular carcinoma (HCC). 83 patients with 100 histologically diagnosed HCCs who underwent pre-operative liver DW imaging with b = 0 and1000 s mm-2 or b = 0 and800 s mm-2 at any of six institutions were included. Two radiologists independently measured the apparent diffusion coefficient (ADC) of the lesion as well as non-ADC parameters, such as the relative contrast ratio and the contrast-to-noise ratio (CNR) between the lesion and the liver parenchyma on high b-value DW images. The diagnostic performance of the DW parameters in discriminating poorly-differentiated HCCs was compared using receiver operating characteristic (ROC) analysis. The areas under the receiver operating characteristic curves for the CNR (86.4% [95% confidence interval (CI) (77.2-95.6] and 83.9% [95% CI 71.2-96.6] for b = 1000 and 800 s mm-2, respectively] and the relative contrast ratio (85.3% [95% CI 75.5-94.8] and 83.5% [95% CI 70.5-96.4]) tended to be superior to the ADC [71.1% [95% CI (56.9-85.2)] and 75.7% [95% CI (55.1-96.2)]; p < 0.05 for CNR vs ADC for b = 1000 s mm-2, but not significant for other parameters) for discrimination of poorly-differentiated HCCs. All DW parameters could discriminate HCC grade. Non-ADC parameters might be more useful than the ADC for predicting poorly-differentiated HCCs. Advances in knowledge: The utility of quantitative DW parameters for predicting HCC grade was demonstrated by using multivendor MR units.