muramidase and Carcinoma--Hepatocellular

Hepatocellular carcinoma (HCC) takes the predominant malignancy of hepatocytes with bleak outcomes owing to high heterogeneity among patients. Personalized treatments based on molecular profiles will better improve patients' prognosis. Lysozyme (LYZ), a secretory protein with antibacterial function generally expressed in monocytes/macrophages, has been observed for the prognostic implications in different types of tumors. However, studies about the explicit applicative scenarios and mechanisms for tumor progression are still quite limited, especially for HCC. Here, based on the proteomic molecular classification data of early-stage HCC, we revealed that the LYZ level was elevated significantly in the most malignant HCC subtype and could serve as an independent prognostic predictor for HCC patients. Molecular profiles of LYZ-high HCCs were typical of those for the most malignant HCC subtype, with impaired metabolism, along with promoted proliferation and metastasis characteristics. Further studies demonstrated that LYZ tended to be aberrantly expressed in poorly differentiated HCC cells, which was regulated by STAT3 activation. LYZ promoted HCC proliferation and migration in both autocrine and paracrine manners independent of the muramidase activity through the activation of downstream protumoral signaling pathways via cell surface GRP78. Subcutaneous and orthotopic xenograft tumor models indicated that targeting LYZ inhibited HCC growth markedly in NOD/SCID mice. These results propose LYZ as a prognostic biomarker and therapeutic target for the subclass of HCC with an aggressive phenotype.

Topics: Animals; Biomarkers, Tumor; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Proliferation; Gene Expression Regulation, Neoplastic; Humans; Liver Neoplasms; Mice; Mice, Inbred NOD; Mice, SCID; Muramidase; Neoplastic Processes; Prognosis; Proteomics

ABIN1, an important immune regulator, has been shown to be involved in various cellular functions, such as immunity, development, tissue homeostasis, and tumor progression. It inhibits TNF- and TLR-induced NF-κB signaling activation and the consequent gene expression. Despite its functional significance, the mechanism of ABIN1 in the regulation of various cellular functions remains unclear. In this study, we identified HDAC1, a key regulator of eukaryotic gene expression and many important cellular events, including cell proliferation, differentiation, cancer and immunity, as an interacting partner of ABIN1. The results showed that ABIN1 acted as a modulator to down-regulate HDAC1 ubiquitination via three different linkages, thereby stabilizing HDAC1 by inhibiting its lysosomal and proteasomal degradation. Interestingly, the inhibitory function of ABIN1 required direct binding with HDAC1. Moreover, the level of p53, which was a tumor suppressor and a well-studied substrate of HDAC1, was under the regulation of ABIN1 via the modulation of HDAC1 levels, suggesting that ABIN1 was physiologically significant in tumor progression. This study has revealed a new function of ABIN1 in mediating HDAC1 modification and stability.

Topics: A549 Cells; Carcinoma, Hepatocellular; DNA-Binding Proteins; Gene Expression Regulation, Neoplastic; Gene Knockout Techniques; HeLa Cells; Hep G2 Cells; Histone Deacetylase 1; Humans; K562 Cells; Liver Neoplasms; Lung Neoplasms; Muramidase; Neoplasms; Proteasome Endopeptidase Complex; Protein Stability; Tumor Suppressor Protein p53; Ubiquitination

The influences of caffeine, lysozyme and the joint application of them on the hepatoma cell line HepG2 proliferation inhibition and cell apoptosis were observed by 3-(4, 5-dimethyl-2-thiazyl)-2, 5-diphenyl-2H-tetrazolium bromide assay and Hoechst 33342, which showed the proliferation inhibition rate of the joint application on HepG2 cells was 47.21%, significantly higher than caffeine or lysozyme, and the joint application promoted the apoptosis of HepG2 cells obviously. Van't Hoff classical thermodynamics formula, the Föster theory of non-radiation energy transfer and fluorescence phase diagram were used to manifest that the process of lysozyme binding to caffeine followed a two-state model, which was spontaneous at low temperature driven by enthalpy change, and the predominant intermolecular force was hydrogen bonding or Van der Waals force to stabilize caffeine-lysozyme complex with the distance 5.86nm. The attenuated total reflection-Fourier transform infrared spectra indicated that caffeine decreased the relative contents of α-helix and β-turn, which inferred the structure of lysozyme tended to be "loose". Synchronous fluorescence spectra and ultraviolet spectra supported the above conclusion. The amino acid residues in the cleft of lysozyme were exposed and electropositivity was increased attributing to the loose structure, which were conducive to increasing caffeine concentration on the HepG2 cell surface by electrostatic interaction to show synergistic effect.

Topics: Apoptosis; Caffeine; Carcinoma, Hepatocellular; Cell Proliferation; Drug Synergism; Hep G2 Cells; Humans; Liver Neoplasms; Muramidase

Immunohistochemical and ultrastructural studies were performed on the sinusoidal lining cells of eight canine hepatocellular carcinomas. The sinusoidal endothelial cells of the tumors had a positive reaction for both Factor VIII-related antigen and peanut agglutinin, but did not bind with Ulex europaeus agglutinin-1. Desmin- and lysozyme-positive cells were present along the sinusoids and perisnusoidal spaces of the tumor tissues, respectively, but were fewer in number compared with those of normal canine liver. Alpha-Smooth muscle actin-positive cells outlining the sinusoids were frequently observed. Electron microscopy revealed that basement membranes were often formed beneath the sinusoidal endothelial cells, with rare fenestration. Macrophages were present around or within the sinusoids and tended to increase in number relative to the degree of tumor differentiation. Myofibroblast-like cells with various morphological features, consistent with alpha-smooth muscle actin-positive cells, were frequently found in the perisinusoidal space. The present study indicates that the sinusoidal lining cells of canine hepatocellular carcinoma have some phenotypic characteristics.

Topics: Actins; Animals; Carcinoma, Hepatocellular; Desmin; Dog Diseases; Dogs; Female; Immunohistochemistry; Lectins; Liver Neoplasms; Macrophages; Male; Microscopy, Electron; Muramidase; Peanut Agglutinin; von Willebrand Factor

Plasma lysozyme levels are elevated in several different pathological conditions. In our study we show that well differentiated human hepatoma cells Hep3B and HepG2 are active synthesis sites of lysozyme and that this synthesis can be modulated by acute phase mediators. The production and modulation of lysozyme synthesis was studied by means of Northern-blot analysis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and a specific bioassay after treatment of the cells with interleukin-1 beta, interleukin-6 and tumor necrosis factor-alpha. Hep3B and HepG2 cells constitutively synthesize high amounts of lysozyme. Lysozyme synthesis and secretion were found to be augmented by interleukin-1 beta and tumor necrosis factor-alpha in both cell lines. Interleukin-6 caused an increase in lysozyme production in Hep3B but a decrease in the HepG2 cells. As expected, the synthesis of albumin was decreased in both cell lines. Furthermore we demonstrated that HepG2 and Hep3B cells produce a biologically active form of the enzyme as measured by a specific bioassay. The results demonstrate that lysozyme is constitutively synthesized by Hep3B and HepG2 hepatoma cell lines and that lysozyme synthesis is modulated by acute-phase mediators. Well differentiated human hepatoma cells may respond differently to different cytokines.

Topics: Albumins; Blotting, Northern; Carcinoma, Hepatocellular; Cytokines; Gene Expression Regulation, Enzymologic; Humans; Interleukin-1; Interleukin-6; Liver Neoplasms; Muramidase; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

Hematoxylin and eosin (H-E) stained liver sections of 47 autopsy cases of hepatic malignancies were examined. There were 43 cases of hepatocellular carcinoma (subtypes of 30 trabecular, 7 solid, 5 pseudoglandular, and one scirrhous carcinoma), 3 of cholangiocellular carcinoma and one of mixed carcinoma. After immunohistochemical staining, benign hepatocytes reacted positively with anti-epithelial membrane antigen (EMA). Hepatocellular carcinoma cells reacted more weakly than benign hepatocytes. It was noted that the microtubular structure, which could not be demonstrated even by alcian blue or cationic ferric hydroxide colloid stabilized with cacodylate (Fe-CaC), was clearly detected with anti-EMA. The EMA-positive microtubular structures may indicate terminal cholangiolar differentiation. Based on EMA, seven more cases formerly classified as hepatocellular carcinoma by H-E were reclassified as mixed carcinoma, totaling eight (17.0%). The histologic classification of "mixed carcinoma" has been 1.5 to 2.0% of primary liver cancers in Japan, but we suggest there may be more cases of "mixed carcinoma" identified in the future. In conclusion, we emphasize that EMA staining is useful for more accurate classification of hepatic tumors.

Topics: Adenoma, Bile Duct; Adult; Aged; Aged, 80 and over; alpha-Fetoproteins; Carcinoma, Hepatocellular; Female; Humans; Immunohistochemistry; Liver Neoplasms; Male; Membrane Glycoproteins; Middle Aged; Mucin-1; Muramidase

The sinusoids of 30 human hepatocellular carcinomas of various types were examined by electron microscopy and histochemically for binding to the Ulex europaeus lectin (UEA1). A population of sinusoidal macrophages was identified with an antibody to lysozyme (muramidase). The UEA1 binding was negative in normal sinusoids but positive in the tumor vessels. Macrophages resembling Kupffer cells were found within the tumor vessels but in smaller numbers than in either normal or cirrhotic liver tissue. Fibrolamellar and sclerosing carcinomas contained the smallest numbers. Ultrastructurally, endothelial cells of tumor vessels were thicker than normal, with fewer fenestrations. They contained bundles of microfilaments and showed basement membrane formation. Subendothelial myoid cells were found. These findings indicate that the sinusoidal vessels of hepatocellular carcinomas show features of true capillaries and precapillary blood vessels. The degree of this difference from normal hepatic sinusoids may reflect the relative immaturity of the cancer cells.

Topics: Adolescent; Adult; Aged; Carcinoma, Hepatocellular; Female; Humans; Immunoenzyme Techniques; Lectins; Liver Neoplasms; Macrophages; Male; Middle Aged; Muramidase; Plant Lectins; Staining and Labeling

We have immunohistochemically localized fibronectin, lysozyme and alpha-fetoprotein (AFP) in 21 human hepatocellular carcinoma (HCC) tissues obtained by surgical resection at both light and electron microscopic levels. Three distinct distribution patterns of fibronectin (sinusoidal, periacinar, and pericellular patterns) were observed. The sinusoidal and periacinar patterns were mainly observed in HCC of pseudoglandular or trabecular patterns and of Edmondson's grade I or II, whereas the pericellular pattern was observed in HCC of compact or trabecular patterns and of Edmondson's III grade, suggesting that the pericellular fibronectin was rather associated with undifferentiated HCC. Electron microscopic observation of the pericellular fibronectin showed fibronectin to be present in the dilated intercellular spaces where microvilli were moderately developed. We observed intracytoplasmic staining of fibronectin in 2 of the 21 HCC cases. By immunoelectron microscopy, fibronectin was observed in the endoplasmic reticulum (ER) of some HCC cells. In the 21 HCC cases, lysozyme-positive cancer cells were observed in 10 cases, and AFP in 6 cases. At the ultrastructural level, lysozyme was identified in the ER and the perinuclear spaces of HCC cells, suggesting that lysozyme was synthesized by these cells. Lysozyme-positive cases tended to be more frequently observed in cases with the pericellular pattern of fibronectin rather than those with sinusoidal or periacinar patterns.

Topics: Adult; Aged; alpha-Fetoproteins; Carcinoma, Hepatocellular; Endoplasmic Reticulum; Female; Fibronectins; Humans; Immunoenzyme Techniques; Liver Neoplasms; Male; Middle Aged; Muramidase

Hepatocellular adenomas are usually visualized as defects on technetium-99m-sulfur colloid liver scans, a fact which has been attributed to the absence of phagocytic Kupffer cells in the tumors. To determine whether this is true, seven hepatocellular adenomas were subjected to immunoperoxidase staining for lysozyme, a marker of mononuclear phagocytes. The Kupffer cells were counted in the tumors and surrounding non-neoplastic liver. All hepatocellular adenomas studied were found to contain Kupffer cells. Three tumors had fewer Kupffer cells than the surrounding liver. Three had about the same number as the surrounding liver, and one had more Kupffer cells than the non-neoplastic liver. Thus, the lack of phagocytosis of colloid in liver scans is probably due to something other than a deficiency of Kupffer cells in the hepatocellular adenomas.

Topics: Adult; Carcinoma, Hepatocellular; Female; Histocytochemistry; Humans; Immunoenzyme Techniques; Kupffer Cells; Liver Neoplasms; Muramidase; Radionuclide Imaging

Topics: Animals; Carcinoma, Hepatocellular; Cell Adhesion; Cells, Cultured; Cytotoxicity, Immunologic; Fibrosarcoma; Guinea Pigs; Liver Neoplasms; Liver Neoplasms, Experimental; Macrophages; Muramidase; Mycobacterium bovis; Phytohemagglutinins; Solubility; Tuberculin

Topics: Animals; Antigens, Neoplasm; Carcinoma, Hepatocellular; Cell Membrane; Chromatography, DEAE-Cellulose; Electrophoresis, Polyacrylamide Gel; Fluorescent Antibody Technique; Galactosidases; Glucosidases; Hyaluronoglucosaminidase; Liver Neoplasms; Muramidase; Neoplasm Proteins; Neoplasms, Experimental; Neuraminidase; Organ Size; p-Dimethylaminoazobenzene; Papain; Peptide Hydrolases; Rats; Solubility; Time Factors; Trypsin