Adjacent Liver Research Articles

Contrast-Enhanced Ultrasound Quantification Assessment of Focal Fatty Variations in Liver Parenchyma: Challenging the Traditional Qualitative Paradigm of Uniform Enhancement With Adjacent Parenchyma.

The purpose of this study was to quantify contrast-enhanced ultrasound enhancement of focal fatty sparing (FFS) and focal fatty infiltration (FFI) and compare it with adjacent liver parenchyma. This was a retrospective observational study yielding 42 cases in the last 4 years. Inclusion criteria were a focal liver lesion, adequate video availability, and an established diagnosis of FFS or FFI based on clinical or imaging follow-up or a second modality. Contrast-enhanced ultrasound examinations were performed with a standard low-mechanical index technique. Commercially available software calculated quantitative parameters for a focal liver lesion and a reference area of liver parenchyma, producing relative indices. In total, 42 patients were analyzed (19 male) with a median age of 18 (interquartile range, 42) years and a median lesion diameter of 30 (interquartile range, 16) mm. The cohort included 26 with FFS and 16 with FFI. Subjectively assessed, 27% of FFS and 25% of FFI were hypoenhancing in the arterial phase, and 73% of FFS and 75% of FFI were isoenhancing. In the venous and delayed phases, all lesions were isoenhancing. The peak enhancement (P = .001), wash-in area under the curve (P < .01), wash-in rate (P = .023), and wash-in perfusion index (P = .001) were significantly lower in FFS compared with adjacent parenchyma but not the mean transit time. In the FFI subgroup, no significant difference was detected. Comparing relative parameters, only the wash-in rate was significantly (P = .049) lower in FFS than FFI. The mean follow-up was 2.8 years. Focal fatty sparing shows significantly lower and slower enhancement than the liver parenchyma, whereas FFI enhances identically. Focal fatty sparing had a significantly slower enhancement than FFI.

A Human Organoid Model of Aggressive Hepatoblastoma for Disease Modeling and Drug Testing.

Simple SummaryHepatoblastoma is the most common childhood liver cancer, making up over 90% of malignant liver tumors in children younger than 5 years of age. Currently, research to find new treatments for treatment-resistant hepatoblastoma is limited by a lack of appropriate models to study the disease. In this study, we describe a novel patient-derived organoid model of aggressive hepatoblastoma that can be used to study the disease in the laboratory and test new treatments. We demonstrate that tumor organoids share the same genomic profile as the patient tumors from which they are derived, and also demonstrate similar features with respect to gene expression profiles and beta-catenin signaling. We also demonstrate the feasibility of using hepatoblastoma organoids to complete a drug screen alongside normal liver control organoids derived from the same patient, and report promising initial results of anti-tumor activity of the BET inhibitor JQ1.Hepatoblastoma is the most common childhood liver cancer. Although survival has improved significantly over the past few decades, there remains a group of children with aggressive disease who do not respond to current treatment regimens. There is a critical need for novel models to study aggressive hepatoblastoma as research to find new treatments is hampered by the small number of laboratory models of the disease. Organoids have emerged as robust models for many diseases, including cancer. We have generated and characterized a novel organoid model of aggressive hepatoblastoma directly from freshly resected patient tumors as a proof of concept for this approach. Hepatoblastoma tumor organoids recapitulate the key elements of patient tumors, including tumor architecture, mutational profile, gene expression patterns, and features of Wnt/β-catenin signaling that are hallmarks of hepatoblastoma pathophysiology. Tumor organoids were successfully used alongside non-tumor liver organoids from the same patient to perform a drug screen using twelve candidate compounds. One drug, JQ1, demonstrated increased destruction of liver organoids from hepatoblastoma tumor tissue relative to organoids from the adjacent non-tumor liver. Our findings suggest that hepatoblastoma organoids could be used for a variety of applications and have the potential to improve treatment options for the subset of hepatoblastoma patients who do not respond to existing treatments.

HMB-45 Negative Adrenal Angiomyolipoma With A Synchronous Adrenal Adenoma – An Unusual Association

Background:&#x0D; Angiomyolipoma (AML) is an uncommon mesenchymal tumour usually found in the kidney. The most common extrarenal site is usually the liver. We report a case of adrenal AML with a predominant vascular component co-existing with an adrenal adenoma.&#x0D; Case report: &#x0D; A fifty-year-old male presented with two months history of dysuria. Computed tomography showed a heterogeneous lesion involving the right adrenal gland abutting the right kidney and adjacent liver capsule. Urine noradrenaline levels were elevated. Adrenalectomy specimen showed an encapsulated mass measuring 8 x 5 x3.5 cm with a lobulated, grey to tan brown cut surface with areas of haemorrhage and tiny cystic spaces. Another yellowish nodule measuring 1.3 x 1.2 x 1 cm was also noted at one pole. Microscopy showed a neoplasm composed predominantly thick-walled vascular channels admixed with foci of smooth muscle bundles and adipocytes. Yellowish nodule showed features of adrenal adenoma. HMB-45 was negative and CD34 positivity highlighted the thick and thin walled vessels.&#x0D; Conclusion: &#x0D; We present the very first case of HMB-45 negative adrenal AML with a predominant vascular component and a synchronous adrenal adenoma.

THE MORPHOGENESIS OF NONPARASITIC LIVER CYSTS

As a result of the introduction of imaging research methods into clinical practice, the frequency of detection of cavities in the liver has significantly increased, among which nonparasitic cysts have not been sufficiently studied. The paper provides an overview of the most commonly used classifications of this pathology. Clinical and radiological manifestations of cysts were investigated based on the analysis of the results of computed tomography, magnetic resonance imaging and ultrasound sonography of 175 patients (109 with solitary cysts and 66 with polycystic liver). The tissues of the resected areas of 81 cysts and adjacent liver tissue were studied by histological and immune-histochemical methods. Various variants of liver cysts were identified: polycystic cysts, solitary cysts and the rarest variant - ciliated anterior-intestinal hepatic cysts. The morphological and histogenetic features of solitary and ciliated anterior-intestinal hepatic cysts, as well as changes in the liver tissue adjacent to the cavities, are described. Polycystic is characterized by the presence of cavities of various sizes, with thin connective tissue partitions and a lining of cubic or flattened epithelium. The lining of solitary cysts is represented by cubic, cylindrical, sometimes multi-row epithelium. Immature biliary structures in the form of von Meijenburg complexes, islets of hepatocytes, conglomerates of dysplastic vessels are revealed in their wall among the cells of connective tissue. Ciliated anterior-intestinal hepatic cysts contain components of the primary intestine wall - a lining of a cylindrical ciliated epithelium, a layer of loose connective tissue, a muscle layer, a connective tissue capsule. Morphological studies made it possible to establish violations of embryogenesis in various types of cysts. Ciliated anterior intestinal hepatic cysts develop from the anterior section of the primary intestine, while solitary cysts are derived from its middle section. Migration of the developing buds of bronchioles from the cranial part of the primary midgut into the hepatic diverticulum determines the presence of all components of the wall of the hollow organ - cylindrical ciliated epithelium, loose connective tissue, bundles of smooth muscle fibers. Simple solitary cysts are the result of abnormal remodeling of the embryonic ductal lamina, thus being a developmental abnormality of the middle section of the primary midgut. Morphological examination of the liver tissue adjacent to the walls of the cysts reveals hypoplasia of lobules, persistence of elements of the embryonic ductal plate in the form of cords and complexes of the biliary epithelium, foci of ductopenia, von Meijenburg complexes, and various variants of vascular dysplasia.

Loss of PTEN expression is associated with PI3K pathway-dependent metabolic reprogramming in hepatocellular carcinoma

BackgroundMetabolic reprogramming, in which energetic metabolism changes from oxidative phosphorylation to glycolysis, is well-accepted as a hallmark of cancers including hepatocellular carcinoma (HCC). A growing body of evidence suggests the involvement of oncogenes and tumor suppressor genes in the control of metabolic reprogramming. In this study, we attempt to investigate whether loss of PTEN, a recognized tumor suppressor, drives metabolic reprogramming of HCC.MethodsCancerous liver tissues were surgically resected from 128 HCC patients, with 43 adjacent noncancerous liver tissues as control. Aerobic glycolysis (Warburg effect) was reflected by measurements of glucose uptake and lactate production, mitochondrial membrane potential collapse was observed by JC-1 staining, glycolytic rate and mitochondrial respiration were evaluated by determining glycolytic proton efflux rate (glycoPER) and oxygen consumption rate (OCR) in cultured human HHCC cells.ResultsReciprocal expression of PTEN and PI3K was determined in cancer liver tissues. Overexpression of PTEN suppressed the Warburg effect, as evidenced by reductions in glucose uptake and lactate production, maintenance of mitochondrial function, and transformation of energetic metabolism from glycolysis to oxidative phosphorylation in cultured PTEN-negative HHCC cells. Importantly, 740 Y-P, a PI3K agonist that leads to activation of the PI3K pathway, partially abrogated the function of PTEN and reprogramed glucose metabolism in cultured HHCC cells.ConclusionsThe discovery that loss of PTEN allows the tumor metabolic program has been a major advance in understanding the carcinogenesis of HCC.Bo7HehjAYW3ADh-U-ymoVNVideo abstractGraphical abstractGraphic abstract showing that loss of PTEN regulates the tumor metabolic program in hepatocellular carcinoma. Loss of PTEN leads to activation of the PI3K pathway enhances the Warburg effect, thereby promoting the development of hepatocellular carcinoma.

Abstract 3750: STEAP2 is upregulated and necessary for hepatocellular carcinoma progression via increased copper levels and stress-activated MAP kinase activity

Abstract To understand potential molecular mechanisms that drive hepatocellular carcinoma (HCC) progression, we performed RNA sequencing from paired adjacent non-tumor liver and HCC tumor tissues of nine local HCC patients which revealed significant involvement of a gene coding for Six Transmembrane Epithelial Antigen of Prostate 2 (STEAP2) protein. STEAP2 is a metalloreductase involved in the reduction and indirectly, uptake, of iron and copper ions. We found significantly higher levels of total copper in HCC tissues than those in paired adjacent non-tumor tissues. The upregulation of STEAP2 expression in HCC was confirmed at its protein level and in TCGA and other published HCC gene expression datasets. Hepatic copper overload is associated with Wilson's disease and a known risk factor for HCC, therefore, we hypothesize that STEAP2 upregulation and copper accumulation contribute to HCC progression. Paired HCC and adjacent non-tumor tissues were collected for RNA sequencing, metal ion measurement, and measurements of STEAP2 levels with RT-qPCR and Western blot. Public HCC datasets were queried for STEAP2 expression in HCC and non-tumor tissues. HCC cell lines with knockdown (KD) and overexpression (OE) of STEAP2 were created to perform mechanistic studies including measurements of copper levels and MAP kinase activities, cell proliferation, migration, and anchorage independent growth in vitro and tumor growth in vivo. STEAP2 is significantly upregulated in various HCC gene expression datasets; its expression is positively associated with tumor grade. STEAP2 KD in HCC cell lines decreased cell growth, migration, invasion, and xenograft tumor growth, while STEAP2 OE showed opposite effects. STEAP2 KD in HCC cells also reduced intracellular copper levels and activation of stress-activated MAP kinases including p38 and JNK. Treatment with copper rescued the reduced HCC cell migration due to STEATP2 KD and activated p38 and JNK. Furthermore, treatment with p38 or JNK inhibitors significantly inhibited cell migration. Thus, STEAP2 appears to play a malignant-promoting role in HCC cells by driving migration/invasion via increased copper levels and MAP kinase activities. Our study uncovered a novel molecular mechanism contributing to HCC malignancy and a potential therapeutic target for HCC treatment. Citation Format: Carla R. Zeballos, Acarizia Easley, Hakim Bouamar, Guixi Zheng, Xiang Gu, Yang Junhua, Yidong Chen, Francisco Cigarroa, LuZhe Sun. STEAP2 is upregulated and necessary for hepatocellular carcinoma progression via increased copper levels and stress-activated MAP kinase activity [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3750.

Abstract 3911: High-throughput screening of patient-derived HCC organoids for drug discovery

Abstract Current in vitro models of primary liver cancer insufficiently recapitulates the tumor heterogeneity and pathophysiology of the original patient's tumor. This significantly hinders translational studies and often leads to the high rate of clinical trial attrition and poor patient response. Recent technological advances in organoid culture methods have been reported to overcome these limitations. Our group has successfully established 3 patient-derived HCC tumor organoids that closely recapitulate the patient's pathophysiology. Conventional short-term monolayer culture of the same 3 tumors as well as 3 non-tumoral adjacent liver and 3 normal liver organoids were established and compared with 5 HCC cell lines from ATCC for their drug response profiles. Subsequent drug screening was performed using a panel of 100 drugs, of which 70 are FDA-approved for either HCC or other cancers, as well as 30 pipeline compounds targeting frequently mutated pathways in HCC. We found that our screening pipeline could achieve robust z' scores and reproducible data points with technical and biological replicates. Principal component analysis of Total Area under curve (AUC) values show distinct and progressive changes between Normal Liver, Non-tumoral adjacent liver, and Tumor organoids. Upon comparison with HCC cell lines Hep3B, HepG2, Huh7, SNU398 and PLC5, we found HCC patient-derived short term cultures, particularly organoids have more diverse drug sensitivity profiles and may be a better indicator of patient heterogeneity. Interestingly, FDA-approved drugs for HCC such as Regorafenib show high sensitivity and tumor specificity to ATCC cell lines (ATCC vs Normal liver P=0.00017***), but this specificity was reduced and lost in patient-derived 2D cultures (P=0.045*) and organoids (P=0.895), respectively. This may explain the observed hepatotoxicity observed in HCC patients and warrant further investigation of more targeted ways of treating HCC. Citation Format: Alissa Michelle Go Wong, Winnie Cheung, Maggie Wong, Aikha Wong, Helena Lam, Zhe Zhang, Matthew Man, Hui Chen, David Close, Paul Johnston, Anthony Chan, Paul Lai, Joseph Kwong, Nathalie Wong. High-throughput screening of patient-derived HCC organoids for drug discovery [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3911.

Novel biomarker for hepatocellular carcinoma: high tumoral PLK-4 expression is associated with better prognosis in patients without microvascular invasion

BackgroundHepatocellular carcinoma (HCC) recurrence after liver resection (LR) adversely affects prognosis but is difficult to predict. Aberrant expression of Polo-Like Kinase 4 (PLK-4) is implicated in several adult malignancies. We sought to evaluate the prognostic value of PLK-4 expression in HCC after curative-intent LR. MethodsPatients undergoing LR for HCC between July-2015 and November-2017 at our centre were retrospectively identified. PLK-4 expression was measured in tumour and adjacent non-tumour liver tissue using quantitative RT-PCR. Disease-free survival (DFS) was evaluated by Kaplan–Meier and Cox proportional hazard models. ResultsA total of 145 patients were identified. Patients were divided according to PLK-4 expression (high: n = 58, low: n = 87) by generating a receiver operating characteristic curve for recurrence with an area under the curve of 0.72 (95% CI: 0.6–0.8). Recurrence and death rates were similar between groups. In patients without mVI, low PLK-4 expression was associated with worse actuarial DFS (low 1-, 3-, 5-year 83%, 60%, 47% vs. high 91%, 81%, 81%; p = 0.02). In patients without mVI, high PLK-4 expression was an independent predictor of survival (HR 0.3, 95% CI: 0.1–1.0; p = 0.04). ConclusionPLK-4 represents a biomarker for good prognosis in patients with HCC who do not have mVI. This could aid clinical decision making for adjuvant clinical trials.

Expression and clinical significance of LAG-3, FGL1, PD-L1 and CD8+T cells in hepatocellular carcinoma using multiplex quantitative analysis

BackgroundFibrinogen-like protein 1 (FGL1)—Lymphocyte activating gene 3 (LAG-3) pathway is a promising immunotherapeutic target and has synergistic effect with programmed death 1 (PD-1)/programmed death ligand 1 (PD-L1). However, the prognostic significance of FGL1-LAG-3 pathway and the correlation with PD-L1 in hepatocellular carcinoma (HCC) remain unknown.MethodsThe levels of LAG-3, FGL1, PD-L1 and cytotoxic T (CD8+T) cells in 143 HCC patients were assessed by multiplex immunofluorescence. Associations between the marker’s expression and clinical significances were studied.ResultsWe found FGL1 and LAG-3 densities were elevated while PD-L1 and CD8 were decreased in HCC tissues compared to adjacent normal liver tissues. High levels of FGL1 were strongly associated with high densities of LAG-3+cells but not PD-L1. CD8+ T cells densities had positive correlation with PD-L1 levels and negative association with FGL1 expression. Elevated densities of LAG-3+cells and low levels of CD8+ T cells were correlated with poor disease outcome. Moreover, LAG-3+cells deteriorated patient stratification based on the abundance of CD8+ T cells. Patients with positive PD-L1 expression on tumor cells (PD-L1 TC+) tended to have an improved survival than that with negative PD-L1 expression on tumor cells (PD-L1 TC−). Furthermore, PD-L1 TC− in combination with high densities of LAG-3+cells showed the worst prognosis, andPD-L1 TC+ patients with low densities of LAG-3+cells had the best prognosis.ConclusionsLAG-3, FGL1, PD-L1 and CD8 have distinct tissue distribution and relationships with each other. High levels of LAG-3+cells and CD8+ T cells represent unfavorable and favorable prognostic biomarkers for HCC respectively.

The Profile and Role of Tumor-infiltrating Lymphocytes in Hepatocellular Carcinoma: An Immunohistochemical Study.

Hepatocellular carcinoma (HCC) is the most common primary malignant tumor of the liver. Tumor-infiltrating lymphocytes (TILs) are a class of cells that form the tumor microenvironment and thus have an effect on carcinogenesis. The aim of this study was to investigate the immunohistochemical expression of CD8, CD4, cytotoxic T lymphocyte-associated protein-4 (CTLA-4), and granzyme B in HCC and their correlation with clinicopathologic parameters and prognosis. This study was carried out on 112 cases of HCC. High percentage of CD8+ TILs was associated with large tumors and adjacent noncirrhotic liver. High percentage of CD4+ TILs and high CD4 to CD8 ratio were associated with nonviral etiology, low alpha fetoprotein, and direct acting antiviral treatment. High percentage of CTLA-4-positive TILs tended to be associated with high-grade HCC, while a high percentage of CTLA-4 in tumor cells was associated with multiple lesions and low tumor grade. High percentage of granzyme B+ TILs was associated with low grade, early stage, and absence of tumor recurrence. High CD4 percentage and high CD4/CD8 ratio affected patients' overall survival. There is a dynamic interaction between the different subsets of lymphocytes in the environment of HCC manifested by coparallel expression of CD4 and CD8 augmenting the expression of CTLA-4, and only CD8 augments the expression of granzyme B. This opens the gate for the beneficial role of immunotherapy in the management of HCC, reducing recurrence and improving survival.

Extracellular ubiquitin promotes hepatoma metastasis by mediating M2 macrophage polarization via the activation of the CXCR4/ERK signaling pathway.

BackgroundStored red blood cell (RBC) transfusion has been shown to enhance the risk of cancer recurrence. However, the underlying mechanism remains unknown. At our lab, we have demonstrated that the extracellular ubiquitin (eUb) released by aged RBCs could promote tumor metastasis in a melanoma mouse model. This study aimed to confirm the pro-tumor effect of eUb on hepatocellular carcinoma (HCC) and explore the related immunoregulatory mechanisms.MethodsForty HCC tissue specimens and the corresponding adjacent nontumor and normal liver tissues were collected. Two human hepatoma cell lines (MHCC-97H and HepG2.2.15), one murine hepatoma cell line (Hepa1-6), and one human monocyte cell line (THP-1) were adopted in this study. The coculture of hepatoma cells with macrophages was initiated with Transwell inserts. Cell migration in vitro was detected by Transwell and wound-healing assays, while in vivo tumor metastasis was measured by luciferase assay and H&E staining. Macrophage polarization was measured by flow cytometry, immunofluorescence, ELISA, qPCR, and Western blot. Protein expression was detected by Western blot, and immunoprecipitation was used to confirm the interaction between Ub and CXCR4 (CXC chemokine receptor type 4).ResultsUb and CXCR4 were significantly upregulated in HCC tissues, and a positive correlation existed between them. In vitro, the migration of hepatoma cells was not affected by eUb directly, but their metastatic abilities were enhanced after coculture with the macrophages pretreated with eUb. Meanwhile, eUb promoted hepatoma cell metastasis in the lung in vivo and increased the ratio of M2 macrophages in the lung tissues and peripheral blood of tumor-bearing mice. Furthermore, the eUb-induced M2 macrophage polarization was related to the activation of the CXCR4/ERK (extracellular regulated protein kinase) signaling pathway.ConclusionsExtracellular ubiquitin promoted hepatoma metastasis through M2 macrophage polarization via the activation of the CXCR4/ERK signaling pathway, indicating that a personalized transfusion strategy is needed for the treatment of HCC patients. Neutralizing Ub in stored RBC units could lessen the detrimental clinical outcomes induced by the transfusion of stored RBCs.

Down-regulation of small nuclear RNA (snRNA) RNU5E-1 in hepatocellular carcinoma presents with vital clinical significance.

For lack of accurate diagnosis and ideal prognosis assessment, hepatocellular carcinoma (HCC) has become the fourth cancers-related death malignant diseases. Small nuclear RNAs (snRNAs) have been investigated as a new class of regulators associated with pathogenesis and clinical evaluation of tumors such as HCC. As for RNU5E-1, one newly identified snRNA, may have similar functions. However, the relationship between RNU5E-1 expression and HCC tumorigenesis remains unclear. The relative RNU5E-1 expression was measured in several HCC cell lines and HCC tissues of 100 patients using quantitative real-time PCR. All patients were grouped according to individual RNU5E-1 expression. Then, the potential association between RNU5E-1 expression in HCC clinical characteristics and prognostic information of patients was evaluated. Compared to human normal hepatocyte cell line QSG-7701, the RNU5E-1 expression in HCC cell lines (fold change: SK-HEP-1, 0.417; Hep 3B, 0.313; Huh-7, 0.189) were significantly down-regulated (P<0.05). Similarly, its expression levels were remarkably lower in HCC tissue than that in corresponding adjacent liver tissues (average fold change: 0.322, P=0.002). Besides, the expression level of RNU5E-1 was remarkably related to tumor size, vessel carcinoma embolus, differentiation level, TNM stages and tumor recurrence rate as well as long-term survival in HCC patients (P<0.05). Moreover, in Kaplan-Meier and Cox regression analysis, RNU5E-1 expression was remarkably correlated to postoperative tumor-free as well as long-term survival in HCC patients as independent factors (P<0.05). The research revealed that RNU5E-1 was down-regulated in HCC and it could be one of indicators for diagnosis and prognostic prediction of HCC patients.

Morphological features of the structure of the parenchyma of the liver depending on the type of the echinococcal cyst

Currently, the problem of echinococcosis remains relevant, since this chronically ongoing and parasitic disease leading to disability occurs in different age groups. In this study, the goal was to conduct a morphological analysis of the fibrous capsule with adjacent liver tissues, depending on the type of echinococcal cysts. The analysis of histological preparations of postoperative biopsies of the liver of 32 patients with echinococcosis after surgery on the liver . A standard morphological analysis of the preparations of the sites of the chitinous membrane and liver parenchyma stained with hematoxylin-eosin and Van Gieson was performed. Types of echinococcal cysts were classified according to the Apoyan-Sarkisyan (1991) classification according to ultrasound sonography and compared with types of cysts according to the international classification of ultrasound images for cystic echinococcosis, which was approved by the WHO. It was found that fibrous membranes in cysts of type II (CE1), III (CE2) and IV (CE3) types may contain germ elements, therefore, this types must be considered as one of the factors for the occurrence of recurrence of echinococcosis.

Contrast-Enhanced Ultrasound Imaging Features of Focal Chemotherapy-Induced Sinusoidal Injury in Patients With Colorectal Cancer: Initial Experience.

Increasing studies have reported focal chemotherapy-induced sinusoidal injury (CSI) mimicking colorectal liver metastasis (CRLM) on imaging studies, resulting in unnecessary lobectomy. This study aimed to investigate the contrast-enhanced ultrasound (CEUS) imaging features of focal CSI. We retrospectively evaluated 16 patients who had a pathologic diagnosis of focal CSI and underwent CEUS between January 2013 and January 2019. The images were compared with those obtained from 27 patients with CRLM. On CEUS, 14 (87.5%) focal CSIs showed heterogeneous isoenhancement, 1 (6.3%) peripheral hyperenhancement, and 1 (6.3%) homogeneous hyperenhancement during the arterial phase. Compared with the adjacent liver parenchyma, the initial enhancement time of focal CSI was earlier in 2 (12.5%), simultaneous in 9 (56.3%), and later in 5 (31.3%) patients. In the portal and late phases, all of the focal CSI cases showed hypoenhancement compared with the adjacent liver parenchyma. Focal CSI had the following CEUS findings, which were different from those of CRLM: (1) heterogeneous isoenhancement in the arterial phase (87.5% versus 0%; P < .001); (2) an initial enhancement time later than that of the liver parenchyma (mean ± SD, 0.5 ± 1.5 versus -1.5 ± 1.9; P < .001); (3) a longer time to peak (30.5 ± 5.6 versus 22.5 ± 4.4 seconds; P < .001); and (4) a later time to wash-out (51.0 ± 12.5 versus 35.0 ± 6.2 seconds; P = .002). Focal CSI usually shows heterogeneous isoenhancement in arterial phase and hypoenhancement in portal and late phases on CEUS, with slow contrast wash-in and wash-out, which can be helpful in the differential diagnosis with CRLM.

Tescalcin is an unfavorable prognosis factor that regulats cell proliferation and survival in hepatocellular carcinoma patients.

BackgroundHepatocellular carcinoma (HCC) is a major health problem and a primary cause of cancer‐related death worldwide. Although great advances have achieved recently by large‐scale high‐throughput analysis, the precise molecular mechanism underlying HCC progression remains to be clearly elucidated. We investigated the relationship between Tescalcin (TESC), a candidate oncogene, and clinicopathological features of HCC patients and explored the role of TECS in HCC development.MethodsTo identify new genes involved in HCC development, we analyzed The Cancer Genome Atlas liver cancer database, and TESC was selected for further investigation. HCC tissue microarray analysis for TESC and its association with clinicopathological features were performed to investigate its clinical significance. TESC was knocked down by using short‐hairpin RNAs. Cell proliferation was analyzed by WST‐1 assay and cell counting. Cell apoptosis was tested by fluorescence‐activated cell sorting. A subcutaneous xenograft tumor model in nude mice was established to determine the in vivo function of TESC. Affymetrix microarray was used to identify its molecular mechanism.Results TESC was significantly increased in HCC tissues compared with the adjacent normal liver tissues. High expression of TESC was detected in 61 of 172 HCC patients by tissue microarray. Large tumor (> 5 cm) and elevated total bilirubin were associated with high TESC expression (both P < 0.050). In multivariate analysis, TESC was identified as an independent prognostic factor for short overall survival of HCC patients. TESC knockdown impaired HCC cell growth in vitro and in vivo. TESC knockdown significantly increased cell apoptosis in HCC cell lines. Furthermore, Affymetrix microarray analysis revealed that TESC knockdown inhibited tumor proliferation‐related pathways while activated cell death‐related pathways.ConclusionTESC was identified as an independent prognostic factor for short overall survival of HCC patients, and was critical for HCC cell proliferation and survival.

The Expression of Dynamin 1, 2, and 3 in Human Hepatocellular Carcinoma and Patient Prognosis.

BackgroundThe classical dynamin family consists of dynamin 1, 2, and 3, which have different expression levels in different tissues to regulate cell membrane fission and endocytosis. Recent studies have reported increased expression of dynamins in human cancer, but their expression in hepatocellular carcinoma (HCC) remains to be determined. This study aimed to investigate the expression of dynamin 1, 2, and 3 in tissue sections of human HCC using quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry.Material/MethodsThe expression of dynamin 1, 2, and 3 were investigated in 192 cases of HCC and 14 paired samples of HCC and adjacent normal liver tissue by qRT-PCR and immunohistochemistry. The clinical significance of dynamin 1, 2, and 3 were determined by correlating their expression levels with patient clinicopathological factors and survival rates. Independent prognostic factors were determined using the Cox regression hazard model.ResultsIn tissue samples from 192 patients with HCC, the expression of dynamin 1, 2, and 3 were upregulated in 41.15%, 29.69%, and 8.33% of cases, respectively. Dynamin 1 had a significantly increased mRNA expression level in HCC compared with adjacent normal liver tissues and was significantly correlated with alpha fetoprotein (AFP) levels, T stage, and TNM stage. Only dynamin 1 expression was correlated with the reduced overall survival (OS), and was identified as an independent prognostic biomarker of human HCC.ConclusionsUpregulation of dynamin 1 at the protein and mRNA level was an independent prognostic biomarker of reduced OS in patients with HCC.

Radiofrequency ablation in management of simple non-parasitic liver cysts

Purpose. Comparison of the long-term outcomes of the treatment of simple liver cysts by traditional and modified RFA methods.Materials and methods. From 2015 to 2018, US-guided RF ablation was used to treat simple nonparasitic hepatic cysts in 14 patients. There were 15 operated cysts in total. 7 patients of group A underwent standard RFA session without temperature restrictions. 7 patients of group B with localization of cysts closely to large vessels and bile ducts or the area of portal fissure, underwent RFA at 60 °C with preinjection of 10% NaCl solution into the cyst cavity. During the intervention, the temperature of the surrounding electrode tissue and the exposure time were recorded.Results. All patients underwent interventions satisfactorily; no complications were observed. In group A, the mean cyst diameter was 50.3 ± 15.2 mm, and the mean initial cyst volume was 77.09 ± 23.2 cm3. In group B, the mean cyst diameter was 61.8 ± 14.9 mm, and the mean initial cyst volume was 137.8 ± 39.2 cm3. After 3 months, the overall decrease in the volume of operated cysts on CT was in group A – 27.1%, in group B – 50.6%; after 6 months – 49.8% and 60.4%, respectively. After 12 months, similar indicators in group A amounted to 64.2%, in group B – 79.2%. 1.5 years after surgery, the volume of operated cysts in group A decreased by 65.2%, in group B by 91.2%; and after 2 years – by 68.9% and 93.6%, respectively. The total decrease of cyst volume at the last control in group B was 24.7% more than in group A. The decrease in cyst volume by more than 50% was noted in group A in 37.5%, in group B – in 85.7%. Complete reduction of the cyst cavity was noted in group A in 25%, in group B in 28.5%. The difference in the decrease of the cyst volumes between the groups after 2 years is statistically significant (p = 0.03), thus it can be concluded that a significantly more pronounced decrease in cyst volumes was observed in group B than in group A.Conclusion. Preinjection into the cystic cavity of a 10% solution of NaCl allowed to achieve a more pronounced decrease of the cystic cavities with the lower RFA temperature regime. At a temperature of 60 °C, the thermal damage minimally spreads to adjacent healthy liver tissue andis sufficient for the radical treatment of the cystic cavity. This makes possible to use the modified RFA method for cysts, located close to the large vessels and bile ducts.

Hsa_circ_0003998 promotes epithelial to mesenchymal transition of hepatocellular carcinoma by sponging miR-143-3p and PCBP1

BackgroundCircular RNAs (circRNAs) play a critical regulatory role in cancer progression. However, the underlying mechanisms of circRNAs in hepatocellular carcinoma (HCC) metastasis remain mostly unknown.MethodsHas_circ_0003998 (circ0003998) was identified by RNAs sequencing in HCC patients with /without portal vein tumor thrombus (PVTT) metastasis. The expression level of circ0003998 was further detected by in situ hybridization on tissues microarray (ISH-TMA) and qRT-PCR in 25 HCC patients with PVTT metastasis. Moreover, the 25 HCC patients with PVTT metastasis and 50 HCC patients without PVTT metastasis were recruited together to analyze the correlation between circ0003998 expression and HCC clinical characteristics. Transwell, migration and CCK8 assays, as well as nude mice model of lung or liver metastasis were used to evaluate the role of circ0003998 in epithelial to mesenchymal transition (EMT) in HCC. The regulatory mechanisms of circ0003998 in miR-143-3p and PCBP1 were determined by dual-luciferase reporter assay, nuclear-cytoplasmic fractionation, fluorescent in situ hybridization, RNA pull- down, microRNA sequence, western blot and RNA immunoprecipitation.ResultsCompared with adjacent normal liver tissues (ANL), circ0003998 expression was significantly upregulated in PVTT tissues and HCC tissues, and its expression correlates with the aggressive characteristics of HCC patients. Further assays suggested that circ0003998 promoted EMT of HCC both in vitro and in vivo. Mechanistically, our data indicated that circ0003998 may act as a ceRNA (competing endogenous RNA) of microRNA-143-3p to relieve the repressive effect on EMT-related stimulator, FOSL2; meanwhile, circ0003998 could bind with PCBP1-poly(rC) binding protein 1 (PCBP1) to increase the expression level of EMT-related genes, CD44v6.ConclusionCirc0003998 promotes EMT of HCC by circ0003998/miR-143-3p/FOSL2 axis and circ0003998 /PCBP1/CD44v6 axis.

MTOR up-regulation of SNRPA1 contributes to hepatocellular carcinoma development.

Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related death worldwide. Recent studies showed that snRNPs were implicated in human cancer development. The role of SNRPA1, which is a member of U2 snRNPs, in HCC, remains undocumented. Here, we found that SNRPA1 was highly expressed in HCC tissue compared with normal adjacent liver tissues. Up-regulation of SNRPA1 was correlated with the clinical stage of HCC and the overall survival of HCC patients. In vitro and in vivo results showed that knockdown of SNPRA1 inhibited the cell proliferation, colony formation and xenografted tumorigenesis of HCC cells. Apoptosis was induced by SNPRA1 down-regulation. Mechanistically, SNPRA1 was stimulated by mTOR activation. In addition, whole-genome microarray analysis identified that 262 genes were up-regulated and 462 genes were down-regulated by SNPRA1 knockdown in HCC cells. qPCR analysis suggested that the fibroblast growth factor-2 (FGF2), Alpha-fetoprotein (AFP), β-catenin, Ki-67 and cyclin B1 were down-regulated and caspase 3, p53 as well as p21 were up-regulated after SNRPA1 knockdown. Taken together, our findings implicate that SNPRA1 functions as an oncogene in HCC.

P0032INCIDENCE OF GALLSTONES AND ASSOCIATED COMPLICATIONS WITH USE OF THE SOMATOSTATIN ANALOGUE LANREOTIDE FOR POLYCYSTIC KIDNEY AND LIVER DISEASE

Abstract Background and Aims Recently, we performed a large randomized-controlled trial to assess the effects of the somatostatin analogue lanreotide on kidney function and kidney and liver volume in autosomal dominant polycystic kidney disease (ADPKD), the DIPAK-1 trial. Lanreotide had no effect on rate of kidney function decline, but slowed kidney and especially liver growth and is used for this indication in clinical practice1,2. A higher incidence of gallstones has been suggested with lanreotide use, but the exact size and implications of this problem are unknown. Therefore, we systematically studied the incidence of gallstones with lanreotide in ADPKD patients. Method In the DIPAK-1 trial, 305 ADPKD patients (age 18-60 years and eGFR 30-60 ml/1.73m2/min) were randomized to lanreotide or standard of care. MRIs were performed at baseline and end of study (120 weeks). For this post-hoc study, patients with a medical history of cholecystectomy (n=7) or absence of a MRI at the end of study (n=28) were excluded. For the remaining study population (n=270), all MRIs were assessed for presence of gallstones by two trained investigators blinded for study treatment and study period and in case of disagreeing results, an abdominal radiologist was consulted for a final assessment. For patients with gallstones additional follow up after the end of the trial was obtained. Results In 2 baseline and 8 end-of -study MRIs, it was not possible to identify the gallbladder, due to multiple adjacent liver cysts. Of the remaining 260 patients (age 48.5 years, female 49.6%, eGFR 50.0 ml/min/1.73m2), 14 patients (5,4%) had gallstones at baseline, of whom 5 were randomized to lanreotide and 9 to standard care. At the end of study, 32 patients (12,3%) had gallstones, of whom 21 patients had received lanreotide and 11 standard care. During the 120 week lasting study, new gallstones occurred in 17 out of 128 patients receiving lanreotide (13.3%) and 3 out of 132 patients receiving standard care (2.3%), rendering an adjusted Odds Ratio for gallstone formation with lanreotide use of 6.9 (95% Confidence Interval 1.9-24.7) p&amp;lt;0.005). The only other significant risk factor for gallstone formation was female gender (OR 3.5, CI 1.2-10.6, p&amp;lt;0.05). Age, body-mass-index and importantly liver volume were not associated with gallstone formation. Of the 32 patients with gallstones at the end of study, seven experienced gallstone-associated complications. Three experienced a biliary pancreatitis (1 during and 2 after the study). Another one had signs of a chronic pancreatitis as accidental finding on imaging during the study, without having had symptoms. Furthermore, another 3 patients underwent a cholecystectomy for symptomatic gallstones after the study. Importantly, all patients with gallstone-associated complications were part of the subgroup of patients that developed de novo gallstones during lanreotide use in the study. Conclusion Lanreotide increases the risk of gallstone formation in ADPKD sevenfold. Gallstones were associated with severe complications, most occurring after the end of the trial. The benefit of somatostatin analogues to reduce kidney and liver size in ADPKD should therefore be weighed against the higher incidence of these adverse events, and patients and doctors should be aware of the risk of (symptomatic) gallstones with use of these drugs.