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Immunohistochemical detection of human telomerase reverse transcriptase in human liver tissues
GASTROENTEROLOGY Vol. 118, No.4
A914 AASLD ABSTRACTS
82
84
IMMUNOHISTOCHEMICAL DETECTION OF HUMAN TELOMERASE REVERSE TRANSCRIPTASE IN HUMAN LIVER TISSUES. Yoshiiku Kawakami. Mikiya Kitamoto, Hiroshi Aikata, Shoichi Takahashi, Toshio Nakanishi, Goro Kajiyama, Wataru Yasui, Jun-ichi Nakayama, Fuyuki Ishikawa, Toshinori Ide, Hiroshima Univ Sch of Med, Hiroshima, Japan; Tokyo Institute of Technology, Japan.
MAJOR LIVER RESECTION INDUCES A SIMILAR DECREASE IN THE FISCHER RATIO AS IN HEPATIC ENCEPHALOPATHY WHICH IMPROVES BY ADMINISTRATION OF BACTERICIDAU PERMEABILITY-INCREASING PROTEIN (RBPI21). Robert 1. Nijveldt, Marinus J. Wiezer, Dirk J. Gouma, Thomas M. Gulik, Inne H. Borel-Rinkes, Hugo W. Tilanus, Camelis 1. Velde, Theo Wiggers, Frans A. Zoetrnulder, Miguel A. Cuesta, Sybren Meijer, Paul A. Leeuwen, Acad Hosp Vrije Univ, Amsterdam, Netherlands; Acad Med Ctr, Amsterdam, Netherlands; Univ Hosp Utrecht, Utrecht, Netherlands; Univ Hosp Digest, Rotterdam, Netherlands; Univ Hosp Leiden, Leiden, Netherlands; Univ Hosp Daniel den Hoed, Rotterdam, Netherlands; The Netherlands Cancer Institute, Amsterdam, Netherlands.
Background/Aims: Human telomerase reverse transcriptase (hTERT) has been identified as a catalytic subunit of telomerase, and is closely correlated with telomerase activity. We have previously demonstrated the significance of telomerase and hTERT mRNA in the human liver tissueespecially in hepatocarcinogenesis (GE 110: A1274, 112: A1344, 114: A1351, 116: A1I81). To clarify the exact location and frequence of teromerase postive cells, we performed in situ immunohistochemical detection of anti-hTERT antibody-reactive proteins in the liver tissue sections. Methods: Fifteen HCC nodules (well-differentiated: 6, moderately differentiated: 7, poorly differentiated:2) and 13 non-cancerous liver tissues (Chronic hepatitis: 6, liver cirrhosis:7) were employed in this study. Telomerase activity was measured by telomeric repeat amplification protocol (TRAP), and expression of hTERT was examined by RT-PCR and immunohistochemistry. Immunohistochemical staining was performed from paraffin embedded sections. After deparaffinization and microwave pretreatment in citrate buffer, the sections were treated with anti-hTERT antibody (anti-hTERT 1.0). The immuno-complex was visualized by a modificaiton of the immunogolobulin enzyme bridge technique (ABC method) using the SensiTek HRP kit (ScyTeck Laboratories, Logan, UT, USA). Since 3. 3'-diaminobezidine tetrahydrochloride was used as substrate, positive reaction was detected as a brown color. Results: HCC cell nuclei were postive to the specific antibody. whereas not to preabsorbed antibody. In well-differentiated HCC, hTERT positive cells were scarcely found, while those in moderately or poorly differentiated HCC were often observed. In non-cancerous Tissues, hTERT positive cells were scarcely observed in hepatocytes and infiltrating lymphocytes. The incidence of hTERT positive cells in well-differentiated HCC was similar to that in non-cancerous liver tissue. The intensity of immunostaining in moderately or poorly differentiated HCC was often higher than that in well-differentiated HCC or non-cancerous liver tissues. Conclusion: This findings suggest that the up-regulation of telomerase according to undifferentiation of HCC may due to the increase of hTERT positive HCC cells and the increase of the expression intensity. Thus, telomerase activity associated with increased hTERT expression may be involved in the development of a majority of hepatocarcinogenesis. 83 EARLY DETECTION OF HEMOBILIA FOLLOWING PERCUTANEOUS ETHANOL INJECTION THERAPY FOR HEPATOCELLULAR CARCINOMA: "HEMOBILIA SIGN" OF GALLBLADDER. Shuntaro Obi, Yasushi Shiratori, Shuichiro Shiina, Keisuke Hamamura, Naoya Kato, Masatoshi Imamura, Takuma Teratani, Shinpei Sato, Yutaka Komatsu, Takao Kawada, Masao Ornata, Univ of Tokyo, Tokyo, Japan. OBJECTIVE Hemobilia often results from an iatrogenic injury caused by therapeutic procedures. This study was carried out to evaluate the efficacy of the ealry diagnosis of hemobilia by ultrasound using 365 consective patient with hepatocellular carcinoma undergoing percutaneous ethanol injection therapy. SUBJECTS AND METHODS Retrospective an prospective study for the early detection of hernobilia to avoid serious following events was conducted from 1995 to 1996. Retrospective analysis using clinical, biochemical and imaging data in 172 patients receiving ethanol injection therapy in 1995 showed that changes in the gallbladder viewed by ultrasonography gave an useful early sign of hemobilia, namely the rapid appearance of echogenic material in the gallbladder lumen. Thus, a prospective study for the early detection of hemobilia was conducted based on this phenomenon in January 1996, and an interventional procedure was halted as soon as the detection of hemobilia. RESULTS The incidence of hemobilia in "the 1996" was not different to that in "the 1995" (8/172 (4.7%) vs 7/193 (3.6%)). However, the mean duration between ethanol injection therapy and the diagnosis of hemobilia was only O.3:tOA days in the "the 1996" compared to 2.8:!::2.1days in the "the 1995" (p
Liver insufficiency as seen in hepatitis and cirrhosis may be accompanied by a hepatic encephalopathy and is characterized by a typical plasma amino acid pattern consisting of a decrease in the Fischer ratio (branched chain amino acids (BCAA)/ aromatic amino acids (AAA)). Little is known about liver resection and its amino acid pattern, although endotoxin plays a major role in the origin of postresection hepatic failure. Therefore plasma amino acid profiles were determined after major liver resection (hemihepatectomy) in patients receiving placebo or the natural endotoxin binder rBPlzl in a phase II clinical trial. Forty-eight patients undergoing major liver resection were randomised in this study, from which 35 patients were evaluable: 18 patients received placebo and 17 patients received rBPlzl (11 patients with high dose rBPl zl , 6 patients with low dose rBPlzl ). Plasma amino acid profiles were determined preoperatively (preop), on the first (dayl) and third (day3) day after the operation. Following major liver resection, Fischer ratio decreased on dayl (p
A914 AASLD ABSTRACTS
82
84
IMMUNOHISTOCHEMICAL DETECTION OF HUMAN TELOMERASE REVERSE TRANSCRIPTASE IN HUMAN LIVER TISSUES. Yoshiiku Kawakami. Mikiya Kitamoto, Hiroshi Aikata, Shoichi Takahashi, Toshio Nakanishi, Goro Kajiyama, Wataru Yasui, Jun-ichi Nakayama, Fuyuki Ishikawa, Toshinori Ide, Hiroshima Univ Sch of Med, Hiroshima, Japan; Tokyo Institute of Technology, Japan.
MAJOR LIVER RESECTION INDUCES A SIMILAR DECREASE IN THE FISCHER RATIO AS IN HEPATIC ENCEPHALOPATHY WHICH IMPROVES BY ADMINISTRATION OF BACTERICIDAU PERMEABILITY-INCREASING PROTEIN (RBPI21). Robert 1. Nijveldt, Marinus J. Wiezer, Dirk J. Gouma, Thomas M. Gulik, Inne H. Borel-Rinkes, Hugo W. Tilanus, Camelis 1. Velde, Theo Wiggers, Frans A. Zoetrnulder, Miguel A. Cuesta, Sybren Meijer, Paul A. Leeuwen, Acad Hosp Vrije Univ, Amsterdam, Netherlands; Acad Med Ctr, Amsterdam, Netherlands; Univ Hosp Utrecht, Utrecht, Netherlands; Univ Hosp Digest, Rotterdam, Netherlands; Univ Hosp Leiden, Leiden, Netherlands; Univ Hosp Daniel den Hoed, Rotterdam, Netherlands; The Netherlands Cancer Institute, Amsterdam, Netherlands.
Background/Aims: Human telomerase reverse transcriptase (hTERT) has been identified as a catalytic subunit of telomerase, and is closely correlated with telomerase activity. We have previously demonstrated the significance of telomerase and hTERT mRNA in the human liver tissueespecially in hepatocarcinogenesis (GE 110: A1274, 112: A1344, 114: A1351, 116: A1I81). To clarify the exact location and frequence of teromerase postive cells, we performed in situ immunohistochemical detection of anti-hTERT antibody-reactive proteins in the liver tissue sections. Methods: Fifteen HCC nodules (well-differentiated: 6, moderately differentiated: 7, poorly differentiated:2) and 13 non-cancerous liver tissues (Chronic hepatitis: 6, liver cirrhosis:7) were employed in this study. Telomerase activity was measured by telomeric repeat amplification protocol (TRAP), and expression of hTERT was examined by RT-PCR and immunohistochemistry. Immunohistochemical staining was performed from paraffin embedded sections. After deparaffinization and microwave pretreatment in citrate buffer, the sections were treated with anti-hTERT antibody (anti-hTERT 1.0). The immuno-complex was visualized by a modificaiton of the immunogolobulin enzyme bridge technique (ABC method) using the SensiTek HRP kit (ScyTeck Laboratories, Logan, UT, USA). Since 3. 3'-diaminobezidine tetrahydrochloride was used as substrate, positive reaction was detected as a brown color. Results: HCC cell nuclei were postive to the specific antibody. whereas not to preabsorbed antibody. In well-differentiated HCC, hTERT positive cells were scarcely found, while those in moderately or poorly differentiated HCC were often observed. In non-cancerous Tissues, hTERT positive cells were scarcely observed in hepatocytes and infiltrating lymphocytes. The incidence of hTERT positive cells in well-differentiated HCC was similar to that in non-cancerous liver tissue. The intensity of immunostaining in moderately or poorly differentiated HCC was often higher than that in well-differentiated HCC or non-cancerous liver tissues. Conclusion: This findings suggest that the up-regulation of telomerase according to undifferentiation of HCC may due to the increase of hTERT positive HCC cells and the increase of the expression intensity. Thus, telomerase activity associated with increased hTERT expression may be involved in the development of a majority of hepatocarcinogenesis. 83 EARLY DETECTION OF HEMOBILIA FOLLOWING PERCUTANEOUS ETHANOL INJECTION THERAPY FOR HEPATOCELLULAR CARCINOMA: "HEMOBILIA SIGN" OF GALLBLADDER. Shuntaro Obi, Yasushi Shiratori, Shuichiro Shiina, Keisuke Hamamura, Naoya Kato, Masatoshi Imamura, Takuma Teratani, Shinpei Sato, Yutaka Komatsu, Takao Kawada, Masao Ornata, Univ of Tokyo, Tokyo, Japan. OBJECTIVE Hemobilia often results from an iatrogenic injury caused by therapeutic procedures. This study was carried out to evaluate the efficacy of the ealry diagnosis of hemobilia by ultrasound using 365 consective patient with hepatocellular carcinoma undergoing percutaneous ethanol injection therapy. SUBJECTS AND METHODS Retrospective an prospective study for the early detection of hernobilia to avoid serious following events was conducted from 1995 to 1996. Retrospective analysis using clinical, biochemical and imaging data in 172 patients receiving ethanol injection therapy in 1995 showed that changes in the gallbladder viewed by ultrasonography gave an useful early sign of hemobilia, namely the rapid appearance of echogenic material in the gallbladder lumen. Thus, a prospective study for the early detection of hemobilia was conducted based on this phenomenon in January 1996, and an interventional procedure was halted as soon as the detection of hemobilia. RESULTS The incidence of hemobilia in "the 1996" was not different to that in "the 1995" (8/172 (4.7%) vs 7/193 (3.6%)). However, the mean duration between ethanol injection therapy and the diagnosis of hemobilia was only O.3:tOA days in the "the 1996" compared to 2.8:!::2.1days in the "the 1995" (p
Liver insufficiency as seen in hepatitis and cirrhosis may be accompanied by a hepatic encephalopathy and is characterized by a typical plasma amino acid pattern consisting of a decrease in the Fischer ratio (branched chain amino acids (BCAA)/ aromatic amino acids (AAA)). Little is known about liver resection and its amino acid pattern, although endotoxin plays a major role in the origin of postresection hepatic failure. Therefore plasma amino acid profiles were determined after major liver resection (hemihepatectomy) in patients receiving placebo or the natural endotoxin binder rBPlzl in a phase II clinical trial. Forty-eight patients undergoing major liver resection were randomised in this study, from which 35 patients were evaluable: 18 patients received placebo and 17 patients received rBPlzl (11 patients with high dose rBPl zl , 6 patients with low dose rBPlzl ). Plasma amino acid profiles were determined preoperatively (preop), on the first (dayl) and third (day3) day after the operation. Following major liver resection, Fischer ratio decreased on dayl (p