Motoi Kondo

Up-regulation of cyclooxygenase-2 in squamous carcinogenesis of the esophagus.

Cyclooxygenase-2 (COX-2) is overexpressed in various types of human malignancies including squamous cell carcinomas (SCCs) of the esophagus, but little is known about COX-2 expression in premalignant esophageal squamous dysplasia. To elucidate the role of COX-2 in esophageal carcinogenesis, we examined the expression of this enzyme in normal squamous epithelium (n = 42), squamous dysplasia [high-grade dysplasia (HGD, n = 41; low-grade dysplasia (LGD, n = 33)]; carcinoma in situ (n = 16), mucosal invasive carcinoma (n = 18), and advanced SCC (n = 45). Immunohistochemistry showed a significantly high COX-2 expression in HGD compared with other lesions. The COX-2 score, an index determined by intensity and positivity of COX-2 staining (maximum 3.0), was 0.29 +/- 0.04 in normal esophagus, 1.75 +/- 0.11 in LGD, 2.89 +/- 0.05 in HGD, 2.17 +/-0.18 in CIS, 1.95 +/- 0.22 in mucosal invasive carcinoma, and 1.81 +/- 0.08 in advanced SCC. Results of reverse transcription-PCR assays confirmed those obtained by immunohistochemistry. COX-2 expression correlated with proliferation activity assessed by the proliferating cell nuclear antigen index in dysplastic lesions (P = 0.001) but not in SCCs. COX-2 expression in SCC did not correlate with various clinicopathological parameters including prognosis. Our results indicate that COX-2 is a sensitive marker for HGD and suggest that COX-2 may be involved in early stages of squamous carcinogenesis of the esophagus.

JTE-522, a cyclooxygenase-2 inhibitor, is an effective chemopreventive agent against rat experimental liver fibrosis1 ☆

BACKGROUND & AIMS The aim of this study was to assess the effects of cyclooxygenase (COX)-2 inhibition on rat experimental liver fibrogenesis. METHODS We investigated the inhibitory effects of a selective COX-2 inhibitor, JTE-522, on liver fibrosis induced by a choline-deficient, l-amino acid-defined diet (CDAA). Inhibitory effect was also tested in a second model of thioacetamide (TAA)-induced liver fibrosis. RESULTS CDAA induced liver fibrosis and preneoplastic foci at 12 weeks and cirrhosis at 36 weeks. Hepatocellular carcinoma was noted in 13 of 15 rats (87%). JTE-522 significantly inhibited fibrosis and development of preneoplastic lesions in a dose-dependent manner and completely inhibited generation of cirrhosis and hepatocellular carcinoma at both low and high doses (10 and 30 mg/kg body wt/day, respectively). JTE-522 administrated only from 12 weeks to 36 weeks also prevented cirrhosis and formation of hepatocellular carcinoma. JTE-522 itself did not cause local or systemic gross or histopathologic changes at 36 weeks. Mechanistic studies indicated that the CDAA model displayed up-regulation of several biomarkers, including COX-2, arachidonate metabolite (prostaglandin E(2)), serum aspartate aminotransferase, and c-myc expression. The model also showed an increased proportion of activated hepatic stellate cells, proliferating cell nuclear antigen index, and CD45-positive inflammatory cells in the liver. JTE-522 effectively diminished these changes. JTE-522 exhibited similar antifibrosis effects in the TAA model. CONCLUSIONS Our results suggest that COX-2 is involved in CDAA- and TAA-induced liver fibrosis. Our data also indicate that JTE-522 is a potent chemopreventive agent of rat liver fibrosis with low toxicity.

Expression pattern of angiogenic factors and prognosis after hepatic resection in hepatocellular carcinoma: importance of angiopoietin‐2 and hypoxia‐induced factor‐1a

Abstract: Background: Hepatocellular carcinoma (HCC) is a hypervascular tumor and angiogenesis plays an important role in its progression. Angiogenesis is regulated by a balance between pro and antiangiogenic molecules. The aim of this study was to investigate the expressions of angiogenic factors and elucidate their roles in angiogenesis in HCC.

Partial Contribution of Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL)/TRAIL Receptor Pathway to Antitumor Effects of Interferon-α/5-Fluorouracil against Hepatocellular Carcinoma

Purpose: Our purpose was to explore the contribution of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)/TRAIL receptor pathway to antitumor effects of IFNα and 5-fluorouracil (5-FU) combination therapy for hepatocellular carcinoma (HCC). Experimental Design: Susceptibility of HCC cell lines to TRAIL and/or 5-FU was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The effects of 5-FU, IFNα, or both on the expression of TRAIL receptors (R1, R2, R3, and R4) on HCC cells or TRAIL in peripheral blood mononuclear cells (PBMC) were examined by flow cytometry. IFNα-induced cytotoxic effects of PBMC on HCC cell lines were examined by 51Cr release assay. TRAIL expression in peripheral blood mononuclear cells and liver tissue from patients was examined by real-time reverse transcription-PCR or immunohistochemistry. Results: HLE and HepG2 were sensitive to TRAIL, but HuH7, PLC/PRF/5, and HLF were resistant. 5-FU had synergistic effect on TRAIL in HLF and additive effect in four other HCC cell lines. TRAIL receptors on HCC cells were up-regulated by 5-FU, and IFNα induced TRAIL on CD4+ T cells, CD14+ monocytes, and CD56+ NK cells. Treatment of effector cells by IFNα and target HCC cells by 5-FU enhanced the cytotoxicity of CD14+ monocytes and CD56+ NK cells against HCC cells via a TRAIL-mediated pathway. TRAIL mRNA overexpression was noted in PBMC of HCC patients who clinically responded to IFNα/5-FU combination therapy, and TRAIL+ mononuclear cells were found in cancer tissue of a responder. Conclusion: Our results suggest that modulation of TRAIL/TRAIL receptor-mediated cytotoxic pathway might partially contribute to the anti-HCC effect of IFNα and 5-FU combination therapy.

Interferon-β Is More Potent Than Interferon-α in Inhibition of Human Hepatocellular Carcinoma Cell Growth When Used Alone and in Combination With Anticancer Drugs

AbstractBackground:The prognosis of advanced hepatocellular carcinoma (HCC) is extremely poor, but promising effects of chemotherapies combined with interferon (IFN) have been reported. Methods:To develop more effective combination therapies for HCC, we compared the antiproliferative effects of IFN-α and IFN-β in combination with various cytotoxic drugs on hepatoma cell lines using MTT assay and isobologram analysis. Results:IFN-β was more potent than IFN-α in inhibiting the cell growth of all cell lines (P < .05, two-way ANOVA). PLC/PRF/5 was more sensitive to either IFN, than HLE and HuH7. Cell growth of all cell lines was inhibited in a dose-dependent manner by 5-fluorouracil (5-FU), cisplatin (CDDP), and doxorubicin (DOX), but the sensitivities of these cells were considerably different. As for IFN-α, synergistic effects were observed when combined with 5-FU and DOX on PLC/PRF/5 cells only, whereas IFN-β showed synergistic effects with 5-FU and CDDP on HuH7 and PLC/PRF/5 cell lines. Conclusion:The spectra of the antiproliferative activity and synergistic effect of IFN-β when combined with anticancer drugs are more potent than those of IFN-α. Combinations of IFN-β and anticancer drugs may provide a better treatment of HCC when combinations with IFN-α are ineffective.

Association Between Recurrence of Hepatocellular Carcinoma and α-Fetoprotein Messenger RNA Levels in Peripheral Blood

PurposeIntra- and extrahepatic recurrence is common, even after curative resection for hepatocellular carcinoma (HCC), suggesting preoperative or intraoperative tumor cell dissemination. Reverse transcription — polymerase chain reaction (RT-PCR) for α-fetoprotein (AFP) is used to detect circulating liver cancer cells. We previously developed a quantitative method that allows estimation of the AFP mRNA level by real-time PCR. In the present study, we used this method to measure the AFP mRNA level before and after resection of HCC, then correlated the findings with various clinicopathological characteristics and prognosis.MethodsWe prospectively examined peripheral blood samples from 38 patients with HCC, and bone marrow aspirate from 25 of these patients. As a control, we examined bone marrow from 20 patients with benign diseases. The follow-up period ranged from 32 to 66 months. Real-time RT-PCR was used to detect AFP mRNA levels in the samples.ResultsAFP was expressed in 9 (23.7%) of the 38 peripheral blood samples. The detection of AFP mRNA was significantly correlated with extrahepatic metastasis after primary surgery, and a shorter disease-free survival time (P = 0.0245 each). Bone marrow samples were defined as positive if they expressed AFP mRNA at levels higher than the maximum expressed level in the controls, because only 1 (5%) of the 20 control bone marrow samples had low AFP mRNA expression. Using this cutoff level, 12 (48%) of the 25 patients with HCC had positivity for AFP mRNA. The results of bone marrow RT-PCR did not correlate with the clinocopathological characteristics of prognosis.ConclusionsUsing real-time PCR to measure the AFP mRNA level in blood, but not bone marrow, could be useful for predicting postoperative tumor recurrence.

Development of a multiple-marker RT-PCR assay for detection of micrometastases of hepatocellular carcinoma.

We investigated the expression of several candidate gene markers: MAGE-1, MAGE-3, cytokeratin-20 (CK-20), and α-fetoprotein (AFP) in tumor tissue and blood specimens from patients with hepatocellular carcinoma to develop a multiple marker reverse transcriptase-polymerase chain reaction (RT-PCR) assay for detection of micrometastasis in circulation. In 24 tumor specimens, the positivity for MAGE-1, MAGE-3, AFP, and CK-20 genes was 71, 67, 88, and 79% respectively, and all specimens expressed at least one marker. Although AFP and CK-20 transcripts were also detected in corresponding noncancerous liver specimens, none of the 22 corresponding normal specimens or seven normal livers were positive for MAGE-1 or MAGE-3 transcripts. In addition, MAGE-1 and MAGE-3 gene transcripts were not detected in any peripheral blood specimens from 31 normal healthy volunteers. MAGE-1, MAGE-3, and AFP transcripts were detected in 9 (12.7%), 3 (4.8%), and 10 (15.9%) of 71 blood specimens from 11 hepatocellular carcinoma patients, respectively, while 19 specimens (26.8%) were positive for at least one marker. Our results indicate that a multimarker RT-PCR assay with cancer-specific markers such as MAGE-1 and MAGE-3 in combination with a liver-specific AFP marker may be a promising diagnostic tool for monitoring hepatocellular carcinoma patients with better sensitivity and specificity.

Successful treatment of multiple hepatocellular carcinoma with tumor thrombi in the major portal branches by intraarterial 5-fluorouracil perfusion chemotherapy combined with subcutaneous interferon-alpha and hepatectomy

We experienced a patient who received successful treatment for multiple hepatocellular carcinoma (HCC) nodules, with tumor thrombi in the major portal branches, with intraarterial 5-fluorouracil perfusion chemotherapy combined with subcutaneous interferon-alpha administration. The patient was a 50-year-old man with hepatitis C virus and HCC. The tumors consisted of a 5-cm main nodule in the right lobe (segment 8) and multiple intrahepatic metastases. The tumor also involved portal vein thrombosis throughout the right portal branch. After two cycles of interferon-alpha/5-fluorouracil combination chemotherapy, tumor markers demonstrated a decreasing tendency. Nine months after the initiation of this therapy, the tumors were limited to the right lobe and were surgically removed by S8 subsegmentectomy, S5 partial hepatectomy, and portal thrombectomy. The serum levels of both alpha-fetoprotein and protein induced by vitamin K absence II fell to normal levels after hepatic resection. Fifty-eight months after the first treatment, he is alive with several recurrent nodules in the liver. In conclusion, the interferon-alpha/5-fluorouracil combination therapy is a useful treatment for HCC in patients who have multiple intrahepatic metastases and portal vein thrombosis. In addition to this therapy, combined modality therapy including, for example, surgical resection, can sometimes have a dramatic therapeutic effect, shown by tumor markers reverting to normal levels.

A Successful Resection and Long-Term Survival of a Patient with Intrahepatic Recurrences of Combined Hepatocellular-Cholangiocarcinoma : Report of a Case

Abstract.Because of the low incidence rate of combined hepatocellular–cholangiocarcinoma (combined HCC-CC), the clinicopathological features of a recurrent tumor of this disease remain to be elucidated. We describe a 47-year-old Japanese woman with a 5-cm diameter mass lesion in the liver. A hepatectomy and dissection of the local lymph nodes were performed and a histological examination of the resected specimen showed combined HCC-CC. After a follow-up of 15 months, intrahepatic recurrence was observed, and a hepatectomy was performed again followed by hepatic arterial infusion chemotherapy. A histological examination revealed that the recurrent tumors consisted of only cholangioacarcinoma. A disease-free survival was obtained for about 7 years after the recurrence. To improve the poor prognosis of combined HCC-CC, clinicopathological features of this disease and the therapy selection for recurrent tumors should be discussed.

Liver Abscess Formation After Microwave Coagulation Therapy Applied for Hepatic Metastases from Surgically Excised Bile Duct Cancer : Report of a Case

We report the case of a liver abscess associated with microwave coagulation therapy (MCT). A 67-year-old man underwent a pylorus-preserving pancreatoduodenectomy for bile duct cancer in August 1998. In June 1999, abdominal computed tomography (CT) showed three low-density areas in his liver. With a diagnosis of multiple metastatic liver tumors, we applied MCT to each liver tumor. The patient presented with fever after therapy. Abdominal CT showed a hepatic abscess, which was treated by percutaneous transhepatic abscess drainage under ultrasonography. A bacteriological examination of the pus material revealed the presence of Enterococcus faecalis and Candida albicans. Treatment resulted in a rapid resolution of symptoms. Our case indicated that care should be exercised in using MCT for the treatment of liver tumors in patients who have undergone reconstructive surgery of the biliary tract.