Association study of the cytochrome P450 gene polymorphisms in Korean patients with hepatocellular carcinoma

Category

OCTREOTIDE

AND OPIOIDS

PROLIFERATION CARCINOMA INCREASE

REDUCE

THE RATE OF

OF HUMAN HEPATOCELLULAR

15DEOXY-D12,14-PGJ2 INDUCTION

cell toxicity, NF-kB suppression or XIAP down-regulation, via a PPARg-independent mechanism in these HCC cells.

appeared to be

(HCC) CELL LINE HEPGS, WITH PARALLEL

Objective: Somatostatin and opioids share similar signaling pathways and their receptors share great homology. We investigated the result of octreotide and opioid treatment on the proliferation rate of HCC cell line HepG2 and on the production of NO by this cell line. Methods: Somatostatin and opioid receptors were detected by PCR. The presence of functional binding sites for somatostatin was further examined by ligand binding assays with the use of radiolabeled Somatostatin. HepG2 cells were cultured with or without the addition of several concentrations of octreotide or opioids and proliferation was measured by tetrazolium salt assay (MTT). Nitrites and Nitrates (end products of NO metabolism) were measured by a calorimetric method which is a modification of the Griess reaction. Results: Octreotide inhibited the proliferation of HepG2 cells with an IC50 of 1 07 x 10m9 M. In parallel, NO release was increased by octreotide (EC50 4,37 x 10m9 M). EKC (6, k, ~1, ~2 receptors ligand), aS1 (K receptor ligand) and DADLE (6 and k receptor ligand) also inhibited HepG2 proliferation (IC.50 2,7 x 10m9 M, 1,64 x lo-” M and 2,62 x lo-’ M respectively) with a concurrent increase of NO release. Conclusions: Octreotide and opioids were found to inhibit the growth of HepG2 cells. This is the first report depicting the direct action of opioids on HCC. In addition, the increase of NO secretion maybe a common pathwayused by octreotide and opioids for their antiproliferative actions. Possible interactions between these two receptor systems and their functional utility in liver biology should be further studied.

346

103

OF NITRIC OXIDE (NO) PRODUCTION

G. Notas’, M. Kampa2, A. Vasilaki3, C. Xidakis’, V. Valatas’, K. Thermos3, G. Kolios’, E. Castanas2, E. Kouroumalis’. Department, Liver Research Laboratory, 2Laboratory ‘Gastroenterology of Experimental Endocrinology, ‘Laboratory Of Pharmacology, University of Crete, School of Medicine, Heraklion, Greece

I

4

REGULATES

AND NF-KB ACTIVATION

INDEPENDENT

MECHANISM

APOPTOSIS VIA A PPARG-

I 347

ASSOCIATION

STUDY OF THE CYTOCHROME

POLYMORPHISMS HEPATOCELLULAR

IN KOREAN

P450 GENE

PATIENTS WITH

CARCINOMA

H.J. Park’, J.I. Lee2, J.H. Jung3, T.H.Han’, D.K. Lee’, S.H. Park’, & Hepatology, J.H. Kim’, W.K. Jang’, C.K. Park’. ‘Gastroenterology Hallym University Sacred Heart Hospital, Anyang, South Korea; 2Gastroenterology & Hepatology, Kyung-Hee University College Of Medicine, Seoul, South Korea; ‘Pharmacology, Kyung-Hee University College Of Medicine, Seoul, South Korea Background: Hepatocellular carcinoma(HCC) is linked etiologically to viruses(HBV and HCV), chemical carcinogens(e.g., aflatoxins), and other environmental factors causing chronic liver injury. Genetic polymorphisms in enzymes involved in carcinogen metabolism may influence susceptibility to cancer. The aim of this study was to examine whether cytochrome P450 2El(CYP 2El) and/or cytochrome P450 lAl(CYP 1Al) genetic polymorphisms were related to susceptibility to HCC. Methods: Genomic DNA from the blood of 79 patients with HCC and 192 control subjects without evidence of cancer, was analyzed by CYP 2El- and CYP lAl-specific PCR, followed by enzyme digestion and gel electrophoresis. Results: There was no significant difference between the patients with HCC and controls in the frequencies of CYP 2El genotypes and alleles with PstI digestion (cl/cl, cllc2, c2/c2 68.8%, 28.6%, 2.6% vs 77.5%, 19.7%, 2.8%). There was no difference between the patients with HCC and controls in the frequencies of CYP 1Al genotypes and alleles with MspI digestion(mUm1, mUm2, m2/m2 47.3%, 32.4%, 20.3% vs 360%, 46.0%, 18.0%). No association with the risk of HCC was observed for the PstI polymorphism of CYP 2El or MspI polymorphism of CYP 1Al according to smoking. No association with the risk of HCC was observed for the PstI polymorphism of CYP 2El or MspI polymorphism of CYP 1Al according to alcohol ingestion. Conclusions: No association with the risk of HCC was observed for the PstI polymorphism of CYP 2El or MspI polymorphism of CYP 1Al.

IN HEPATOCELLULAR

CARCINOMA

H. Okano, K. Shiraki, H. Inoue, T. Kawakita, Y. Saitou, N. Enokimura, N. Yamamoto, M. Deguchi, K. Sugimoto, T. Sakai, K. Murata, T. Nakano. First Department Of Internal Medicine, School Of Medicine, Mie University, Tsu, Japan The peroxisome proliferator-activated receptor-g (PPARg) -high affinity ligand, 15deoxy-D-12, 14.PGJ2 (15d-PGJ2), is toxic to malignant cells through cell cycle arrest or apoptosis induction. In this study, we investigated the effects of 15d-PGJ2 or apoptosis induction or expression of apoptosis related proteins in hepatocellular carcinoma (HCC) cells. 15dPGJ2 induced apoptosis in SK-Hepl and HepG2 cells at 50 microM. Pretreatment with the Pan-caspase-inhibitor, benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethyl ketone (Z-VAD-fmk) only partially blocked apoptosis induced by 15d-PGJ2 at 40 microM. This indicated that 15d-PGJ2 induction of apoptosis is associated with a caspase-3-independent pathway. 15d-PGJ2 also induced down-regulation of X-chromosome-linked inhibitor of apoptosis (XIAP), Bclx and Apaf-1 in SK-hepl cells, but not in HepG2. However 15d-PGJ2 sensitized both HCC cell lines to TRAILinduced apoptosis. In SK-Hepl cells, cell toxicity, Nl-kB suppression and XIAP down-regulation were induced by 15d-PGJ2 treatment under conditions where PPARg was down-regulated. These results suggest that the effect of 15d-PGJ2 was through a PPARg-independent mechanism. Although cell toxicity was induced when PPARg was down-regulated in HepG2 cells, Nl-kB suppression and XIAP down-regulation were not induced. In conclusion, 15d-PGJ2 induces apoptosis of HCC cell lines via caspasedependent or -independent pathways. The abilities of 15d-PGJ2 to induce

I

348

PROSPECTIVE

ASSIGNEMENT

NON-RESECTABLE DIFFERENT

OF PATIENTS WITH

HEPATOCELLULAR

CARCINOMA

TO

THERAPIES

R.R. Plentz’, K.L. Rudolph’, J. Bleck’, M. Gebel’, T. Kirchhoff2, S. Kubicka’ , M.P. Manns I. ’Gastroenterology, Hepatology And Endocrinology, Medical School Hannoves Hannoves Get-many; 2Radiology, Medical School Hannoves Hannoves Get-many Introduction: Hepatocellular carcinoma (HCC) is the most common type of primary liver carcinoma in developed countries. Most patients present at a stage when surgical resection is no longer possible. As non-surgical therapies there are number of locoregional treatments, including transcatheter arterial chemoembolisation (TACE), percutaneous ethanol injection (PEI) and systemic therapy for example with octreotide. Material and Methods: 150 patients with inoperable HCC were prospectively assigned to different therapies using following criteria: 1. TACE followed of PEI with locally limited tumor and sufficient liver function (CHE > 2.0, CHILD A-B, Quick > 50%, Prothrombin activity < 50 set, platelet count > 50000, creatinin activity < 100 kmol/l and unblocked portal venous system). 2. Exclusive PEI with locally limited tumor and TACE exclusion details. 3. Exlusive TACE with disseminated multilobular tumor and TACE including details. 4. Octreotide therapy with disseminated multilobular tumor and insufficient liver function. Results: 44 patients were treated with PEI, 33 patients were treated with TACE, 32 patients were treated with TACE and PEI, and 41 patients were treated with octreotid. We observed no incidence of severe side effects