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Hepatocellular carcinoma with chronic B-type hepatitis complicated by autoimmune hemolytic anemia:A case report

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World Journal of Gastroenterology 2007年第32期13卷 4401-4404页

作者： toshie okada Keiichi Kubota Junji Kita Masato Kato Tokihiko Sawada Department of Gastro-enterological Surgery Dokkyo University School of MedicineTochigiJapan

A 57-year-old man consulted a local hospital because of a persistent slight fever. At the age of 37 years he was diagnosed having B-type hepatitis, but left the liver dysfunction untreated. Twenty years later, he was diagnosed having chronic hepatitis B, hepatocellular carcinoma （HCC） and macrocytic anemia, and referred to our hospital for further investigation. A HCC with a maximum diameter of 5.2 cm was detected in segment 8. Results of blood tests included 1.8 mg/dL serum total bilirubin, 0.9 mg/dL bilirubin, less than 10 mg/dL haptoglobin, 7.9 g/dL hemoglobin, 130 fL MCV, and 14.5% reticuloo/tes. A bone marrow sample showed erythroid hyperplasia. The direct Coombs test gave a positive result. We diagnosed the anemia as autoimmmune hemolytic anemia （AIHA）, for which prednisolone could not be administered due to positivity for HBsAg and HBeAg. After preparation of washed blood cells for later transfusion, the patient underwent systematic resection of segment 8. The cut surface of the resected specimen demonstrated an encapsulated yellow-brownish tumor measuring 52 mmx 40 mm which was diagnosed pathologicaly as moderately differentiated HCC. On the 9th postoperative day, the patient＇s temperature rose to 38℃, and exacerbated hemolysis was observed. The maximum total bilirubin value was 5.8 mg/dL and minimum hemoglobin level was 4.6 g/dL. He tolerated this period without blood transfusion. Currently he is being followed up as an outpatient, and shows no signs of HCC recurrence or symptoms of anemia. AIHA associated with HBV infection has been described in only three previous cases, and the present case is the first in which surgery was performed for accompanying HCC.

关键词： Hepatocellular carcinoma B-type hepatitis Auto-immune hemolytic anemia

MK615 inhibits pancreatic cancer cell growth by dual inhibition of Aurora A and B kinases

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World Journal of Gastroenterology 2008年第9期14卷 1378-1382页

作者： toshie okada Tokihiko Sawada Tatsushi Osawa Masakazu Adachi Keiichi Kubota Second Department of Surgery Dokkyo University School of Medicine Kitakobayashi 880MibuShimotsugaTochigi 321-0293Japan Second Department of Surgery Dokkyo University School of Medicine Kitakobayashi 880MibuShimotsuga Tochigi 321-0293Japan Japan Apricot Co.Ltd.1-11-7 MidorichoTakasakiGunma 370-0073 Japan Japan Apricot Co. Ltd.1-11-7 MidorichoTakasakiGunma 370-0073 Japan Second Department of Surgery Dokkyo University School of MedicineKitakobayashi 880Mibu ShimotsugaTochigi 321-0293Japan

AIM:To investigate the anti-neoplastic effect of MK615, an anti-neoplastic compound isolated from Japanese apricot, against human pancreatic cancer cells in vitro. METHODS: Three human pancreatic cancer cell lines PANC-1, PK-1, and PK45H were cultured with MK615 at concentrations of 600, 300, 150, and 0 μg/mL. Growth inhibition was evaluated by cell proliferation assay, and killing activity was determined by lactate dehydrogenase (LDH) assay. Expression of Aurora A and B kinases was detected by real-time polymerase chain reaction (PCR) and Western blotting. Cell cycle stages were evaluated by flow cytometry. RESULTS: The growth inhibitory rates of MK615 at 150, 300, and 600 μg/mL were 2.3% ± 0.9%, 8.9% ± 3.2% and 67.1% ± 8.1% on PANC1 cells, 1.3% ± 0.3%, 8.7% ± 4.1% and 45.7 ± 7.6% on PK1 cells, and 1.2 ± 0.8%, 9.1% ± 2.1% and 52.1% ± 5.5% on PK45H cells, respectively (P <0.05). The percentage cytotoxicities of MK615 at 0, 150, 300, and 600 μg/mL were 19.6% ± 1.3%, 26.7% ± 1.8%, 25.5% ± 0.9% and 26.4% ± 0.9% in PANC1 cells, 19.7% ± 1.3%, 24.7% ± 0.8%, 25.9% ± 0.9% and 29.9% ± 1.1% in PK1 cells, and 28.0% ± 0.9%, 31.2% ± 0.9%, 30.4% ± 1.1% and 35.3 ± 1.0% in PK45H cells, respectively (P < 0.05). Real-time PCR and Western blotting showed that MK615 dually inhibited the expression of Aurora A and B kinases. Cell cycle analysis revealed that MK615 increased the population of cells in G2/M phase. CONCLUSION: MK615 exerts an anti-neoplastic effect on human pancreatic cancer cells in vitro by dual inhibition of Aurora A and B kinases.

关键词： Japanese apricot Aurora kinases Pancreatic cancer Cell cycle

New anti-proliferative agent,MK615,from Japanese apricot “Prunus mume” induces striking autophagy in colon cancer cells in vitro

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World Journal of Gastroenterology 2007年第48期13卷 6512-6517页

作者： Shozo Mori Tokihiko Sawada toshie okada Tatsushi Ohsawa Masakazu Adachi Kubota Keiichi Second Department of Surgery Dokkyo University School of Medicine Japan Apricot Co. Ltd

AIM: To investigate the anti-neoplastic effects of MK615, an extract from the Japanese apricot (Prunus mume), against colon cancer cells. METHODS: Three colon cancer cell lines, SW480, COLO, and WiDr, were cultured with MK615. Growth inhibition was evaluated by cell proliferation assay and killing activity was determined by lactate dehydrogenase assay. Induction of apoptosis was evaluated by annexin Ⅴ flow cytometry. Morphological changes were studied by light and electron microscopy, and immunofluorescence staining with Atg8. RESULTS: MK615 inhibited growth and lysed SW480, COLO and WiDr cells in a dose-dependent manner. Annexin Ⅴ flow cytometry showed that MK615 induced apoptosis after 6 h incubation, at which point the occurrence of apoptotic cells was 68.0%, 65.7% and 64.7% for SW480, COLO, and WiDr cells, respectively. Light and electron microscopy, and immunofluorescence staining with Atg8 revealed that MK615 induced massive cytoplasmic vacuoles (autophagosomes) in all three cell lines. CONCLUSION: MK615 has an anti-neoplastic effect against colon cancer cells. The effect may be exerted by induction of apoptosis and autophagy.

关键词： Colon cancer Japanese apricot Prunus mume Autophagy Apoptosis MK615

MK615 decreases RAGE expression and inhibits TAGE-induced proliferation in hepatocellular carcinoma cells

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World Journal of Gastroenterology 2010年第42期16卷 5334-5341页

作者： Yuhki Sakuraoka Tokihiko Sawada toshie okada Takayuki Shiraki Yoshikazu Miura Katsuya Hiraishi Tatsushi Ohsawa Masakazu Adachi Jun-ichi Takino Masayoshi Takeuchi Keiichi Kubota Second Department of Surgery Dokkyo Medical UniversitySchool of MedicineKitakobayashi 880MibuShimotsugaTochigi 321-0293Japan Public Health Dokkyo Medical UniversitySchool of MedicineKitakobayashi 880MibuShimotsugaTochigi 321-0293Japan AdaBio Co.Ltd.21-1 Kenzaki-machiTakasakiGunma 370-0883Japan Department of Life Pharmacy Faculty of Pharmaceutical SciencesHokuriku UniversityHo-3Kanagawa-machiKanazawaIshikawa 920-1181Japan

AIM:To investigate the proliferative effect of advanced glycation end-products(AGEs) and the role of their cellular receptor(RAGE) on hepatocellular carcinoma(HCC) cells,and the inhibitory effects of MK615,an extract from Japanese apricot,against AGEs were also ***:Two HCC cell lines,HuH7 and HepG2,were *** of RAGE was investigated by poly-merase chain reaction,Western blotting,and flow cytemetry(FACS).The effect of MK615 on RAGE expression was also evaluated by *** proliferative effects of a control(unglycated bovine serum albumin),glucosederived AGEs(Glc-AGE),and glyceraldehyde-derived AGEs(Glycer-AGE),and the anti-proliferative effect of MK615 against AGEs,were evaluated using MTT ***:Expression of RAGE was confirmed at both the mRNA and protein levels in both HuH7 and *** revealed that the level of RAGE expression was higher in HuH7 than in *** with 0.1 μg/mL MK615 decreased the expression level of RAGE from 24.3% to 3.7% in HuH7 and from 6.2% to 4.8% in *** growth indices for the control,Glc-AGE,and Glycer-AGE were 1.06 ± 0.08,0.99 ± 0.04,and 1.38 ± 0.05,respectively,in HuH7(P = 0.037),and were 1.03 ± 0.04,1.04 ± 0.03,and 1.07 ± 0.05,respectively,in HepG2(P > 0.05).When the cells were cultured simultaneously with Glycer-AGE and MK615,MK615 abrogated the proliferative effect of Glycer-AGE in ***:Only Glycer-AGE has a proliferative effect on HuH7,which expresses a higher level of ***615 suppresses the proliferative effect of GlycerAGE on HuH7 by decreasing the expression of RAGE.

关键词： Hepatocellular carcinoma Receptor of advanced glycation end-product Advanced glycation endproducts MK615 Toxic advanced glycation end-products