chondroitin-sulfates and Carcinoma--Hepatocellular

Chronic liver diseases have both high incidence and mortality rates; therefore, a deeper understanding of the underlying molecular mechanisms is essential. We have determined the content and sulfation pattern of chondroitin sulfate (CS) and heparan sulfate (HS) in human hepatocellular carcinoma and cirrhotic liver tissues, considering the etiology of the diseases. A variety of pathological conditions such as alcoholic liver disease, hepatitis B and C virus infections, and primary sclerosing cholangitis were studied. Major differences were observed in the total abundance and sulfation pattern of CS and HS chains. For example, the 6-O-sulfation of CS is fundamentally different regarding etiologies of cirrhosis, and a 2-threefold increase in HS N-sulfation/O-sulfation ratio was observed in hepatocellular carcinoma compared to cirrhotic tissues.

Topics: Carcinoma, Hepatocellular; Chondroitin Sulfates; Glycosaminoglycans; Heparitin Sulfate; Humans; Liver Cirrhosis; Liver Neoplasms; Pilot Projects

Abnormal expression of chondroitin sulfate has been found in many types of cancer, while its biological functions in hepatocellular carcinoma (HCC) progression remain uninvestigated. Here, we report that chondroitin sulfate synthase 1 (CHSY1), the enzyme that mediates the polymerization step of chondroitin sulfate, is a critical mediator of malignant character in HCC that acts via modulating the activity of the hedgehog signaling. CHSY1 was up-regulated frequently in HCC where these events were associated with worse histologic grade and poor survival. Enforced expression of CHSY1 was sufficient to enhance cell growth, migration, invasion, and epithelial-mesenchymal transition, whereas silencing of CHSY1 suppressed these malignant phenotypes. Mechanistic investigations revealed that the increase of cell surface chondroitin sulfate by CHSY1 promoted sonic hedgehog binding and signaling. Inhibiting hedgehog pathway with vismodegib decreased CHSY1-induced migration, invasion, and lung metastasis of HCC cells, establishing the critical role of hedgehog signaling in mediating the effects of CHSY1 expression. Together, our results indicate that CHSY1 overexpression in HCC contributes to the malignant behaviors in cancer cells, we provide novel insights into the significance of chondroitin sulfate in hedgehog signaling and HCC pathogenesis.

Topics: Anilides; Animals; Carcinoma, Hepatocellular; Cell Movement; Cell Proliferation; Chondroitin Sulfates; Epithelial-Mesenchymal Transition; Female; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Glucuronosyltransferase; Hedgehog Proteins; Hep G2 Cells; Humans; Liver Neoplasms; Lung Neoplasms; Male; Mice, Inbred C57BL; Middle Aged; Multifunctional Enzymes; N-Acetylgalactosaminyltransferases; Neoplasm Grading; Neoplasm Invasiveness; Phenotype; Pyridines; RNA Interference; Signal Transduction; Transfection; Xenograft Model Antitumor Assays

In this study, we demonstrated a novel polyelectrolyte microparticle, doxorubicin(DOX)-superparamagnetic iron oxide (SPIO)-chondroitin sulfate (CS)/chitosan (CHI)microparticles (MPs), as a drug delivery system for hepatic cancer treatment. We also investigated the properties of these microparticles through composition determination, formulation tests, in vitro study, and in vivo study. The results showed that our DOX-SPIO-CS/CHI MPs had an average diameter of 1.43±0.54μm and exhibited a spherical shape. The encapsulation efficiency of this drug carrier was approximately 31±8.07%, according to our spectroscopic determination. The results of release profile test revealed the sustained-release behavior of DOX-SPIO-CS/CHI MPs, which released 51.5% of DOX within 48h of the testing period. According to the results of a cell viability assay and an animal study, the DOX-SPIO-CS/CHI MPs exhibited stronger cytotoxicity than did free DOX when it was administered to Hep G2 and Huh-6 human liver cancer cell lines in vitro and to nude mice of Hep G2/Huh-6-bearing mice model in vivo.

Topics: Animals; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Survival; Chitosan; Chondroitin Sulfates; Doxorubicin; Drug Carriers; Female; Ferric Compounds; Hep G2 Cells; Humans; Liver Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Microscopy, Electron, Transmission; Microspheres; Neoplasm Transplantation; Particle Size; Polysaccharides; Spectrophotometry

The characteristics of glycosaminoglycans (GAGs) in many carcinomas have been reported to be different from those in normal tissues, which can be used as prognostic indices in some cancers. However, the difference in GAG characteristics among various differentiation status or histological types of the same cancer has not been described. The aim of this study was to investigate the relationship between GAG characteristics and human primary hepatic carcinomas of divers differentiation status or histological type.. GAGs from intrahepatic cholangiocarcinomas and differently differentiated hepatocellular carcinomas were extracted, purified and enzymatically digested. Their content, relative molecular size distribution and disaccharide composition were analyzed and compared using electrophoresis and high-performance liquid chromatography.. A progressive increase in the content of chondroitin sulfate, low molecular size GAGs, and nonsulfated and disulfated chondroitin sulfate disaccharide units, together with a gradual decrease in heparan sulfate, have been found as the differentiation status of hepatocellular carcinoma became poorer. A significant increase in hyaluronic acid, which only slightly increased in hepatocellular carcinoma, was found in intrahepatic cholangiocarcinomas.. The alterations in GAG characteristics in primary hepatic carcinoma were associated with both the differentiation status and the histological type of the tumor.

Topics: Bile Duct Neoplasms; Bile Ducts, Intrahepatic; Biomarkers, Tumor; Carcinoma, Hepatocellular; Cell Differentiation; Cholangiocarcinoma; Chondroitin Sulfates; Glycosaminoglycans; Humans; Liver Neoplasms

Synthesis of apolipoprotein (apo)E in hepatocytes leads to both secretion and retention in cell surface pools. Inclusion of Brefeldin A to HepG2 cells prompted a rapid decrease of cell surface apoE to about 37% of control values after a 3-h incubation. The t(1/2) for this dynamic pool was estimated to be 15 min. In contrast, a stable fraction of apoE (t(1/2) > 20 h) was found in association with the extracellular matrix (ECM). Increased content of apoE on the ECM correlated with decreased binding of VLDL. Decreased apoE on the cell surface correlated with increased binding of VLDL to cells. Collectively, this suggests that glycosaminoglycan-bound apoE can occlude binding sites for apoE-containing lipoproteins on glycosaminoglycans. In solid-phase assays, heparin, suramin, and chondroitin sulfates A and B efficiently inhibited the binding of apoE to heparan sulfate proteoglycans, but were unable to displace apoE from this glycosaminoglycan. Finally, decreasing cell surface apoE with suramin subsequently decreased the apoE content on secreted apoB-containing lipoproteins without affecting the overall secretion of apoE or apoB to the extracellular medium. In summary, cell surface apoE comprises both dynamic fractions, which can be donated to newly secreted lipoproteins, and stable fractions, which may act to minimize the unproductive binding of lipoproteins to the ECM.

Topics: Apolipoproteins E; Binding Sites; Brefeldin A; Carcinoma, Hepatocellular; Cell Membrane; Chondroitin Sulfates; Dermatan Sulfate; Extracellular Matrix; Glycosaminoglycans; Half-Life; Heparin; Humans; Kinetics; Lipoproteins; Lipoproteins, VLDL; Liver Neoplasms; Suramin; Tumor Cells, Cultured

The optimum conditions for iron colloid enhanced fast spin echo (FSE) in the detection of hepatocellular carcinoma have not been clearly established as yet. MRI was performed on 14 patients with hepatocellular carcinoma (45 nodules) before and after administration of chondroitin sulphate iron colloid (CSIC). One type of conventional spin echo (CSE) (TR/TE = 1800/80) was then quantitatively and qualitatively compared with three types of FSE (FSE 1800 (TR/effective TE/echo factor = 1800/90/7); FSE 7 (3500/90/7); and FSE 11 (3500/99/11)). The liver signal-to-noise ratio (SNR) was significantly decreased after CSIC administration in all sequences, while the tumour-to-liver contrast-to-noise ratio (CNR) was significantly increased. Although the decreased ratio of the liver SNR was smaller on the three FSE sequences compared with CSE, the increased ratio of the tumour-to-liver CNR was higher on the FSE sequences. The highest increase of the tumour-to-liver CNR was on the FSE 7 sequence. The number of detectable tumours, both before and after the administration of CSIC, was largest on FSE 7. In conclusion, FSE with longer TR and TE, and decreased echo factor, was especially useful for CSIC enhanced liver MRI.

Topics: Adult; Aged; Carcinoma, Hepatocellular; Chondroitin Sulfates; Colloids; Contrast Media; Female; Humans; Iron; Liver Neoplasms; Magnetic Resonance Imaging; Male; Middle Aged; Sensitivity and Specificity

Time-course changes in contrast enhancement of chondroitin sulphate iron colloid (CSIC), an MR contrast agent, were determined in 12 patients with 20 lesions of classical hepatocellular carcinoma (HCC). Spin echo T1 weighted (T1WI) and T2 weighted images (T2WI) were obtained before administration of CSIC and 1, 6 and 24 h after injection. The signal-to-noise ratio (SNR) in the tumour region and the liver and the tumour-to-liver contrast-to-noise ratio (CNR) were calculated, and time-course changes of these ratios were determined. SNRs for tumour before the administration of contrast medium did not differ significantly from SNRs after administration on T1WI or T2WI. SNRs for the liver on both T1WI and T2WI were significantly lower at each time point after administration than before administration. The tumour-to-liver CNRs for both T1WI and T2WI were significantly higher after administration than before administration. The maximum CNR was observed 6 h after administration on both T1WI and T2WI. The contrast enhancement was maintained for at least 24 h, with a peak at 6 h after administration. The prolonged enhancement obtained with CSIC has extended the time during which effective contrast is maintained.

Topics: Aged; Carcinoma, Hepatocellular; Chondroitin Sulfates; Colloids; Contrast Media; Humans; Image Enhancement; Iron; Liver Neoplasms; Magnetic Resonance Imaging; Male; Middle Aged; Time Factors

To demonstrate the usefulness of iron colloid-enhanced MR images in differentiating hepatocellular carcinoma (HCC) from hyperplastic nodules (HN), microautoradiographs of chemically induced rat liver tumours were prepared 4 h after intravenous injection of chondroitin sulphate iron colloid (CSIC) labelled with 59Fe by the dipping technique. 20 Wistar rats were allocated into three groups: (1) a normal group, 10; (2) an HN group, 5; and (3) a liver cancer (LC) group, 5. In the I.C group, a diet containing 0.06% 3'-methydiaminobenzine tetrahydro-chloride (DAB) was administered for 3 months. In the HN group, a diet containing 0.025% acetylaminofluorene (AAF) was administered for 4 months. Non-labelled CSIC was intravenously injected into five rats in the normal group, and pseudomicroautoradiographs were prepared using the same technique (normal cold group). 50 sites for examination were randomly selected for each of the normal liver tissue, HN, well-differentiated HCC (HCC-W), and moderately to poorly-differentiated HCC (HCC-MP). The number of Kupffer cell-like macrophages and the photosensitized area ratio (PAR) per field of view were calculated. There was no significant difference in either the number of Kupffer cell-like macrophages or the PAR between HN and normal liver tissue. Although there was no significant difference in the number of these cells between groups HN and HCC-W, the PAR in group HCC-W was significantly lower than that in group HN (p = 0.045). In HCC-MP, both their number (p = 0.003) and the PAR (p = 1.18 x 10(-9)) were significantly lower than in group HCC-W. However, the PAR in HCC-MP was significantly higher than those in the normal cold group (p = 0.019). Iron colloid-enhanced MRI is useful for differentiating HCC from HN, and for diagnosing the degree of HCC differentiation.

Topics: Animals; Carcinoma, Hepatocellular; Chondroitin Sulfates; Colloids; Diagnosis, Differential; Hyperplasia; Liver Neoplasms; Magnetic Resonance Imaging; Rats; Rats, Wistar

Dual contrast magnetic resonance imaging (DCMR) with combined use of a negative contrast medium, chondroitin sulfate iron colloid (CSIC), and a positive contrast medium, Gd-DTPA, was attempted in 20 cases of hepatocellular carcinoma. Spin echo T1 weighted and T2 weighted images (T1WI, T2WI), and T1 weighted images 15 min after intravenous injection of Gd-DTPA (0.1 mmol kg-1) were obtained. Within 1 week, these MR studies were repeated within 1 h of intravenous injection of CSIC (23.6 mumol Fe kg-1) under similar conditions. DCMR and the other five imaging techniques were visually evaluated and compared in terms of tumour detectability, tumour spread and qualification of tumours (depiction of inner structure). DCMR was significantly better than Gd-DTPA enhanced T1WI in tumour detectability, and better than Gd-DTPA enhanced T1WI or CSIC enhanced T1WI in depicting tumour spread. In the qualification of tumours, DCMR was significantly better than all the other five imaging techniques. None of the patients in this study showed adverse reactions or significant changes in biochemistry. DCMR is an imaging technique which is able to utilize the characteristics of these contrast agents collectively, and exhibits advantages in grasping the inner structure of tumours, especially in hepatocellular carcinoma.

Topics: Carcinoma, Hepatocellular; Chondroitin Sulfates; Colloids; Contrast Media; Drug Combinations; Female; Gadolinium DTPA; Humans; Iron; Liver Neoplasms; Magnetic Resonance Imaging; Male; Meglumine; Middle Aged; Organometallic Compounds; Pentetic Acid

The association between contrast enhancement by chondroitin sulfate iron colloid (CSIC) and the histological grade of hepatocellular carcinoma (HCC) was evaluated in 24 patients diagnosed by histological examination of surgical specimens (26 nodules: 11 well-differentiated and 15 poorly-moderately-differentiated nodules). In the well-differentiated HCC nodules, the tumor-liver contrast to noise ratio (CNR) was not significantly increased after i.v. CSIC injection on both T1-weighted and T2-weighted images. In the moderately-poorly-differentiated HCC, CNR was significantly increased after CSIC administration on both T1-weighted and T2-weighted images (p < 0.01). MR imaging using CSIC may be useful for diagnosing the degree of HCC differentiation.

Topics: Carcinoma, Hepatocellular; Chondroitin Sulfates; Contrast Media; Female; Humans; Kupffer Cells; Liver; Liver Neoplasms; Magnetic Resonance Imaging; Male; Middle Aged

Chondroitin sulfate iron colloid (CSIC) was used as an MR contrast agent for the detection of hepatocellular carcinoma (HCC). The findings of 25 surgically confirmed HCCs in 19 patients were retrospectively analyzed. T1-, T2- and proton density-weighted spin echo MR images were obtained before and after i.v. injection of 23.6 microM Fe/kg of CSIC. Unenhanced and CSIC-enhanced MR images and images obtained by CT during arterial portography (CT-AP) were correlated with surgical pathology findings. The sensitivities of CSIC-enhanced and unenhanced MR imaging, and CT-AP were 92%, 80%, and 88%, respectively. No significant differences were noted. Portal flow abnormalities demonstrated by CT-AP did not affect the detection of HCC by CSIC-enhanced MR imaging. CSIC-enhancement at MR imaging was a disadvantage in the detection of lesions less than 1 cm in diameter. CSIC-enhanced MR imaging is a supplemental method for the detection of HCC.

Topics: Carcinoma, Hepatocellular; Chondroitin Sulfates; Colloids; Contrast Media; Female; Humans; Iron; Liver Neoplasms; Magnetic Resonance Imaging; Male; Middle Aged; Portography; Retrospective Studies; Tomography, X-Ray Computed

Using a 1.5 T MR imaging unit, T1- and T2-weighted images were obtained before and after i.v. administration of chondroitin sulfate iron colloid (CSIC) in order to differentiate hepatocellular carcinoma (n = 20) from adenomatous hyperplasia without atypia (n = 16). Differentiation was made from the tumor-liver contrast to noise ratio (CNR) and visual evaluation of the nodule, with reference to signal intensity relative to that of the surrounding liver. The CNR of adenomatous hyperplasia was on T1-weighted images significantly decreased after CSIC administration (p < 0.01). On T2-weighted images, there was no significant difference in CNR after CSIC administration. On the other hand, the CNR of hepatocellular carcinoma was significantly increased after CSIC administration on both T1- and T2-weighted images (p < 0.01). CSIC reflects intratumor reticuloendothelial cellular functions, and is therefore useful in differentiating hepatocellular carcinoma from adenomatous hyperplasia without atypia.

Topics: Carcinoma, Hepatocellular; Chondroitin Sulfates; Colloids; Contrast Media; Female; Humans; Iron; Liver Neoplasms; Magnetic Resonance Imaging; Male; Middle Aged; Precancerous Conditions

The inhibitory effect of sulfated polysaccharides on blood-borne metastasis was examined. As a model of blood-borne metastasis, the ascitic form of hepatoma AH-109A tumor was injected intravenously into Donryu strain rats. Examination of the pulmonary metastatic nodules developed 2 weeks later showed inhibitory effect of the five sulfated polysaccharides tested. Xylan sulfate was the most inhibitory, and exerted its inhibitory effect when the tumor cells were in the pulmonary capillary beds. However, fromthe rapid disappearance of radioactivity from the lungs after injection of 125IUDR-labeled AH-109A cells, tumor cells seemed to be retained in the lungs for only a very short time. Measurement of the anticoagulative and fibrinolytic activities of three sulfated polysaccharides showed that the inhibitory effect of these compounds on blood-borne metastasis was proportional to their anticoagulative and fibrinolytic activities, xylan sulfate showing the highest activities. These results suggest that sulfated polyaccharides may inhibit blood-borne pulmonary metastasis by inhibiting the lodging of tumor cells in the pulmonary capillary beds.

Topics: Animals; Blood Coagulation; Carcinoma, Hepatocellular; Chondroitin Sulfates; Female; Fibrinolysis; Idoxuridine; Lipoprotein Lipase; Liver Neoplasms; Lung Neoplasms; Neoplasm Metastasis; Polysaccharides; Rats; Sulfuric Acid Esters