Eiki Matsushita

Clinical Pilot Study of Intrahepatic Arterial Chemotherapy with Methotrexate, 5-Fluorouracil, Cisplatin and Subcutaneous Interferon-Alpha-2b for Patients with Locally Advanced Hepatocellular Carcinoma

To evaluate the efficacy of methotrexate (MTX)-5-fluorouracil (5-FU), cisplatin (CDDP), and interferon-α-2b(IFNα-2b) combination therapy, we conducted a clinical pilot study in patients with locally advanced hepatocellular carcinoma (HCC). Sixteen patients, who had received no prior treatment for the HCC, with portal tumor thrombosis in the main trunk or in the major branch were enrolled in the study. IFNα-2b (3 × 106 units) was injected subcutaneously 3 times per week. After a bolus administration of MTX (30 mg/m2), CDDP (75 mg/m2) and thereafter 5-FU (750 mg/m2) were given weekly by intrahepatic arterial infusion. In 15 eligible patients, there were 1 complete response (CR) and 6 partial responses (PR) with a response rate of 46.7%. Median survival of the 15 patients was 7 months, and the 2-year survival rate of CR and PR patients was 57.1%. There was severe transient hematologic toxicity. More than grade 2 nausea/vomiting was noted in >50%. In conclusion, the IFNα-2b combination chemotherapy demonstrated good response in patients with locally advanced HCC. This treatment should be tried in a controlled study.

Expression of interferon alpha/beta receptor in the liver of chronic hepatitis C patients

Interferon (IFN) demonstrates antiviral activity by binding to receptors on the cell surface. Expression of the IFN receptor in hepatocytes may be directly associated with a hepatitis C virus (HCV) infection and the response to IFN therapy. A competitive PCR method was developed to measure IFN alpha/beta (α/β) receptor mRNA in liver samples obtained by needle biopsy. Thirty‐one patients with chronic hepatitis C (21 without cirrhosis, 10 with cirrhosis) and six normal subjects were used. Eighteen of the 21 patients without cirrhosis received the IFN therapy. Competitive PCR was carried out using IFN α/β receptor gene‐specific primers and a specific competitor. Expression of the receptor was detected in all liver samples. There was no association between the expression level and serum alanine aminotransferase level, serum (2′–5′) oligo (A) synthetase level, amount of serum HCV RNA, or HCV genotype. The expression level in patients with chronic hepatitis was significantly higher than that in normal livers (P < 0.05) and in cirrhotic livers (P < 0.01). Seven of the 18 patients treated with IFN demonstrated a sustained response to IFN (sustained responders), and the remaining 11 did not (nonsustained responders). The expression level of IFN α/β receptor mRNA in the sustained responders was significantly higher than that in the nonsustained responders (P < 0.01). Thus, the expression of IFN α/β receptor mRNA may be one of the host factors influencing the response to IFN therapy. J. Med. Virol. 56:217–223, 1998.

Involvement of IL-10, an anti-inflammatory cytokine in murine liver injury induced by Concanavalin A

A mechanism of liver injury such as, viral hepatitis or autoimmune hepatitis is considered to involve the impairment of hepatocytes mainly mediated by T-cell immunity, but the roles of a variety of cytokines involved in regulation remain unclarified. We investigated the involvement of various cytokines, particularly, interleukin-10 (IL-10) which is considered to be an anti-inflammatory cytokine, in a murine model of experimental liver injury induced by Concanavalin A (Con A). The model of liver injury was made by intravenous injection of Con A (0.5 &mgr;g) through the caudal vein in 6-week-old female BALB/c mice weighting 20 g. By collecting blood before and at 1, 3, 6, 12 and 24 h after the injection of Con A, alanine aminotransferase (ALT) levels were sequentially measured, and liver tissue was sampled to examine liver injury. Furthermore, TNF-alpha, IL-4 and IL-10 levels were sequentially determined by enzyme-linked immunosorbent assay (ELISA). Serum ALT significantly increased between 3 and 24 h after the Con A injection, and spotty necrosis was histologically observed, suggesting mild liver injury. TNF-alpha and IL-4 increased soon after the injection of Con A. IL-10 increased bimodally soon after and at 12 h after the Con A injection. After neutralizing antibodies (1 &mgr;g) to IL-10 were intraperitoneally injected into the same model at 6 h before Con A treatment, serum ALT levels and the histology of the liver were examined 12 h after the Con A injection. ALT was significantly higher in the group treated with anti-IL-10 antibody (130.7+/-33.5 IU per I) than in the non-treated group (56.5+/-3.5 IU per I) (P<0.05). Histological examination showed spotty necrosis in the group treated with anti-IL-10 antibody. These results suggest that IL-10 has inhibitory effect on liver injury in a murine model of Con A-induced experimental liver injury mediated by cellular immunity.

Upregulation of Type I Interferon Receptor by IFN-gamma

Type I interferon (IFN) receptor has a multichain structure composed of at least two distinct subunits, IFNAR-1 and IFNAR-2. In the present study, we demonstrated that IFN-gamma induced the expression of mRNA for IFNAR-1 and IFNAR-2 in a human hepatoma cell line, HepG2 cells. The induction was dose and time dependent. Because of this result, we examined the effect of combined treatment with type I IFN and IFN-gamma. The intracellular 2-5A-synthetase activity induced by combined treatment was significantly higher than that by type I IFN alone. This study suggests that combined treatment with type I IFN and IFN-gamma may be more effective than that of type I IFN alone and that the upregulation of type I IFN receptor may be one of the reasons. Our findings may have some relevance to the clinical use of IFN.

A case of progressive multiple focal nodular hyperplasia with alteration of imaging studies

Focal nodular hyperplasia (FNH) of the liver is a lesion characterized by a well circumscribed region of hyperplastic liver tissue with stellate fibrosis. The pathogenesis of the lesion is unknown but various authors consider that FNH may be a response to a preexisting vascular abnormality. We experienced a case of progressive multiple FNH, in which the hemodynamic change as shown by imaging modalities, may support this hypothesis. The patient, a 38-yr-old woman, was found by chance to have multiple portal venous shunts and multiple FNH in both lobes of her liver. Because of their benign characteristics, we followed the nodules periodically without any special treatment. After about 4 yr, the nodules increased both in size and number. In addition, digital subtraction angiography showed that the diameter of the artery had become larger. The hemodynamic change revealed by imaging studies in this case supports the hypothesis that one of the pathogens of FNH is a secondary hepatocellular response to arterial hyperperfusion caused by some vascular malformations.

The Role of Hepatitis C Virus in Hepatocellular Carcinoma in Japan

Increasing numbers of patients with hepatocellular carcinoma (HCC) have been reported in Japan. In this paper, we investigated the role of hepatitis C virus (HCV) in HCC and the reason for the increase, using patients admitted to our university hospital from 1945 to 1992. 99 (73%) of 135 patients with HCC were positive for anti-HCV. Prospective studies demonstrated that 22 of 158 (14%) patients with chronic hepatitis C, and 31 of 70 (44%) cirrhotic patients with anti-HCV developed to HCC during the follow-up period (10.1 +/- 3.3 and 7.3 +/- 3.5 years, respectively). Prolonged survival of cirrhotic patients during past decades would also contribute to the increasing number of HCC cases as well as the number of HCV infections in Japan.

Gastrointestinal amyloidosis secondary to hypersensitivity vasculitis presenting with intestinal pseudoobstruction

Hypersensitivity vasculitis is characterized byinflammation and necrosis of small blood vesselssecondary to allergic or hypersensitivity mechanisms (1,2). Gastrointestinal involvement with edema and bleeding also has been reported (3-5).Long-standing inflammation, such as rheumatic disease,infectious disease, inflammatory bowel disease, familialMediterranean fever, and malignancy, may lead tosystemic amyloidosis (6). Gastrointestinal involvementmay induce anorexia, nausea and vomiting, diarrhea,constipation, bleeding, malabsorption, andpseudoobstruction (6, 10-12). In this report we discussa patient with hypersensitivity vasculitis with severeintestinal bleeding who developed systemic amyloidosiswith intestinal pseudoobstruction 29 months after onset.Secondary amyloidosis due to hypersensitivity vasculitis has not been previously reported,and the causal relationship is discussed in thisreport.

Interferon-alpha modulates resistance to cisplatin in three human hepatoma cell lines

Abstract: We investigated the expression of the drug resistance-related genes, multidrug resistance gene 1 (MDR1), multidrug resistance associated protein gene (MRP), and the DNA topoisomerase IIα, DNA topoisomerase IIβ, and glutathione-S-transferase π gene (GST-π) in three human hepatoma cell lines (HepG 2, HuH 7, SK-Hep-1) with or without drug treatment with interferon-α (IFN-α) and cisplatin (CDDP), by a reverse transcription-polymerase chain reaction (RT-PCR) method and a competitive PCR method. The signals of the MDR1, MRP, topoisomerase IIα, and topoisomerase IIβ genes in HepG2 were weakened when IFN-α was added to CDDP. In SK-Hep-1, the administration of CDDP alone increased the signals of MDR1 while the addition of IFN-α decreased the signals, and the signals of GST-π were decreased by IFN-α plus CDDP. In summary, our results concerning the expression of drug resistance-related genes in three human hepatoma cell lines demonstrate that IFN-α may modulate the mechanism of resistance to CDDP in liver cancer.

Floating density of hepatitis C virus particles and response to interferon treatment

Hepatitis C virus (HCV) particles can be classified into two major fractions according to their floating density in serum. However, the genomic heterogeneity of each fraction and the relationship between this viral characteristic and interferon (IFN) response in patients with chronic hepatitis are not known. In this study, floating density and single strand conformation polymorphism (SSCP) of the hypervariable region 1 (HVR1) of HCV were examined in 16 patients with chronic hepatitis prior to IFN treatment. The ratio of HCV RNA titers in the top (T) and bottom (B) fractions, or T:B ratio, was 10:1 in 4 patients, 1:1 in 7, and 1:10 in 5. Three of the 4 patients with a 10:1 ratio showed a sustained response to IFN, while none of the 5 patients with a 1:10 ratio demonstrated a sustained response (P < 0.05). All 4 patients with a 10:1 ratio had 1 or 2 SSCP bands, and 4 of the 5 patients with a 1:10 ratio had 4 or 5 bands (P < 0.01). Furthermore, the number of SSCP bands in the top fraction from 6 sustained responders (1.8 ± 0.3) was significantly smaller than from 10 non sustained responders (4.1 ± 0.8) (P < 0.05). Thus, patients with a high T:B ratio and low heterogeneity in HVR1 demonstrated sustained responses to IFN, while those with low T:B ratios and high heterogeneity did not. J. Med. Virol. 55:12–17, 1998.

Importance of achieving complete necrosis during the first treatment for hepatocellular carcinoma to prevent bone metastasis: A prospective study

Background and Aims: Recent advances in the treatment of hepatocellular carcinoma (HCC) have changed the importance of bone metastasis during the follow up of such patients. In the present study, we investigated risk factors for bone metastasis after treatment for HCC.