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Coexpression of transforming growth factor alpha and epidermal growth factor receptor in human hepatocellular carcinoma
A1098
AASLD ABSTRACTS
OCOMPARISON BETWEEN THE EFFECTS OF INTERFERON a AND INTERFERON fl ON LIPID METABOLISM. K.Kioka, K.So, Y.Moriyoshi, H.Nebiki, t(Okawa, H.Oka, H.Yamada, S.Harihara, *T.Kuroki, *K.Kobayashi. Department of Gastroenterology, Osaka City General Hospital and "Third Department of Internal Medicine, Osaka City University Medical School, Osaka, Japan There have been reports sug~esling that interferon(IFN) affects lipid metabolism in animals. However, despite extensive clinical use of IFN in treatment of chronic hepatitis and malignant tumors, changes in lipid metabolism associated with IFN administration to humans have been only minimally studied. In addition, no comparison has been made of the effects of IFN alpha( a ) and IFN beta(/3 ) on lipid metabolism. In this study, 44 patients with chronic hepatitis C were divided into a group treated with IFN a ( a group, 22 patients) and one treated with IFN /3 (/3 group, 22 patients), and changes in lipid metabolism during IFN therapy were evaluated. In the c~ group, the serum free fatty acid level (FFA) increased significantly (p<0.01.) from 0.29±0.16 mEq/l before IFN administration to 0 . 4 2 + 0 . 2 3 mEq/1. during administration. In the fl group, sis0, it increased significanfly (p<0.01) from 0.35 ± 0.18 mEq/1 before IFN administration to 0.55 ± 0.34 mEq/1 during administration. I n t h e a group, the serum triglyceride level (TG) increased significantly (p<0.01) from 110.0 ± 52.3 mg/dl before IFN administration to' 130.5 ± 48.1 mg/dl during administration. In the/3 group, also, it increased significantly (p<0.01) from 127.0 ± 68.7 mg/dl before administration to 298.4 ± 267.7 mg/dI during administration. However, no significant changes were observed in serum total cholesterol level (T-CHO) or serum glucose level during IFN administration in either group. Conclusions. These fIndings suggest that IFN administration increases serum TG and FFA in patients with chronic hepatitis C. No difference in the effects on I!pid metabolism was observed between the a group and fl group. Since IFN administration markedly increase serum TG in some patients, and hypertriglyceridemia of more than 1000mg/dl is associated with a high risk of pancreatitis, careful observation of patients receiving IFN will be required for increase in serum TG and other side effects.
SERUM LEVELS OF TRANSFORMING GROWTH FACTOR ALPHA IN PATIENTS WITH CHRONIC HEPATITIS C. S.Kira,T.Nakanishi,R.Nakashio,M.Kitamoto,Y.Watanabe, G.Kajiyama,S.Suemori*.First Dept. of Internal Medicine,Hiroshima University School of Medicine,Hiroshima,Japan.*Gastrointestinal Unit,Yokohama City Seibu Hospital,St.Marianna University School of Medicine, Yokohama,Japan.
Previous studies have indicated that transforming growth factor alpha (TGF-alpha) play an important role in liver regeneration and hepatocarcinogenesis.We also have reported that serum TGF-alpba levels was el~va~l in patients with chronic hepatitis and hepatocellular carcinoma.Aim:The aim of this study was to assess the changes of serum TGF-alpha levels and efficacy of interferon (IFN) therapy in patients with chronic hepatitis C. Materials and Methods:Using enzyme-linked immunosorhent assay kit (Oncogene S~ienee Co.),we studied the serum 'levels of TGF-alpha (before,at the end of and 6 months after IFN therapy) in 14 patients with chronic hepatitis C who had been pathologically diagnosed as chronic aggresive hepatitis(CAH)2a by aspiration biopsy and had undergone IFN therapy.Seven patients with complete response had maintained normal ALT value for over 6 months after IFN therapy.And seven patients with non response had maintained abnormal high ALT value during the follow up. Results: In non response cases, the mean (+SD) TGF-alpha levels was 283.27 +32.27pg/ml before therapy,296.81+--29.64 pg/ml at the end of therapy and 284.87+42.36 pg/ml 6 months after therapy.In contrast,in Complete response cases,the mean (+SD) TGF-alpha levels was 209.80 +68.86 pg/ml before therapy,152.28+ 65.16 pg/ml at the end of therapy and 117.32+ 68.12 pg/ml 6 months after therapy.in complete response cases,serum TGF-a!pha levels at the conclusion of and 6 months after IFN therapy were lower than that before IFN therapy(P<0.05).There was significant statistical correlation between serum TGF-alpha le~,els and Serum ALT values. Conclusion:These results Suggest that serum TGF alpha levels may reflect the degree of liver damage and TGF-alpha may play a role in liver regeneration.
GASTROENTEROLOGY,Vol. 108, No. 4
COEXPRESSION OF TRANSFORMING GROWTH FACTOR ALPHA AND EPIDERMAL GROWTH FACTOR RECEPTOR IN HUMAN HEPATOCELLULAR CARCINOMA. S,Kira,R.Nakashio,M.Kitamoto,Y.Watanabe,T.Nakanishi, G.Kajiyama. First Dept. of Internal Medicine,Hiroshima University School of Medicine,Hiroshima,Japan. Previous studies have indicated that transforming growth factor alpha (TGF-alpha) acts on epidermal growth factor receptor (EGFR) and play an important role in liver regeneration and hepatocarcinogenesis.We aslo have reported the elevation of serum TGF-alpha levels in patients with hepatocellular carcinoma(HCC) by enzyme-linked immunosorbent assay. Aim: To compare the immunoaclfivity with the proliferation and the degree of historogic differentiation of HCC,the coexpression of TGF-alpha and EGFR was examined in the same area of HCC by immunohistochemical studies.Materials and Methods: Specimens of 48 nodules of HCC less than 5cm in diameter(17:well differentiated,25:moderately differentiated, 6:poorly diffe~rentiated ) were obtained from 44 Japanese patients who had undergone surgical resection.Immunoreactiv e TGF-alpha and EGFR in the same area of paraffin-embedded sections were detected by using monoclonal antibody(Oneogene Science Co.,Bio Genex Co.). Results:In adjacent non tumorous tissue,TGF-alpha was weakly expressed in all cases and EGFR was expresse'd in only 10 of 48(20.8%)cases.The expression of TGF-a!pha and EGFR in HCC was as follows; " TGF-alpha EGFR well (17) moderately (25) poorly (6) total (÷) • (+) 6/17 7/25 1/6 (14) (+) (-)' 7/17 7•25 1/6 (15) (+) (+) 0/17 3/25 4/6 ( 7) ('4") (:) 4/17 8'/25 0/6 (12) ( + )~: strongly 15ositive,(+) : weakly positive, (-) : negative In the cases Showing strongly positive staining of TGF-alpha,EGFR was expressed at almost equal frequency.Otherwise,in the cases showing weakly positive staining of TGF-alpha,the expression of EGFR differs statistically ~ccording to the degrees of histologic differentiation of HCC(p<0.05).In comparison with well or morerately differenciated HCC,poorly differentiated HCC expresses EGFR highly with the enhaficed proliferative responce to weak TGF-alpha expression.Regarding the tumor diameter and cumulative recurrence rates,significant difference was found between EGFR-positive groups and EGFR-negative groups(p<0.05). Conclusion: These results suggest that TGF-alpha / EGFR may play a role in the differentiation and the proliferation of HCC
TRANSFORMING GROWTH FACTOR ALPHA ANTISENSE DNA OLIGONUCLEOTIDES APPROACH TO INHIBIT GROWTH OF HUMAN HEPATOCELLULAR CARCINOMA DERIVED CELL LINES. S.Kira,T.Nakanishi,R.Nakashio,M.Kitamoto,Y.Watanabe, G.Kajiyama,S.Suemori*.First Dept. of Internal Medicine,Hiroshima University School of Medicine'Himshima'Japan'*Gastr°intestinal Unit,Yokohama City Seibu Hospital,St.Marianna University School of Medicine, Yokohama,Japan.
Previous studies have indicated that transforming growth factor alpha (TGF-alpha) play an important role in liver regeneration and hepatocarcinogenesis.We also have reported that serum TGF-alpha levels was elevated in patients with chronic hepatitis and hepatocellular carcinoma(HCC). Aim:To better understand the role of TGF-alpha in development of HCC,we investigated the specific inhibition of TGF-alpha gene by TGF-alpha DNA ann-sense posphorothioate oligonucleotides (SODN,Oncogene Science Co.) in HCC derived cell lines. Materials and Methods: 1) Human HCC derived cells (Hep G2,HLF)were plated at 1.0xl04cells/well in 96 well plates and grown in 100gl of WEM supplemented with 10% FCS.Human TGF-alpha SODN and control oligonucleotides were added directly into the medium 48hrs after plating cells.3H thymidine incorporauon was measured 24hrs after addition.2)TGF-alpha in culture medium was measured by using enzyme-linked immunosorbant assay kit (Oncogene Science Co.). Results: TGF-alpha in culture medium was 6.71 _l.57pg/24hr/g protein (HepG2), 4.99 +-1.58pg/24hr/g protein (HLF).3H thymidine incorporation ratio to non addition controls was as follows; TGF-alpha S-ODN Control oligonucleotides 0.5 gtM 1.01.tM 2.0p.M 0.5 ~tM 1.0gM 2.0p.M HepG2 96% 76% 72% 1 0 8 % 106% 94% HLF 96% 82% 89% 125% 110% 140% Conclusion:These results suggest that TGF-alpha could be an autocrine factor in hepatocellular carcinoma derived cell lines and TGFalpha may play a role in development of hepatocellular carcinoma.
AASLD ABSTRACTS
OCOMPARISON BETWEEN THE EFFECTS OF INTERFERON a AND INTERFERON fl ON LIPID METABOLISM. K.Kioka, K.So, Y.Moriyoshi, H.Nebiki, t(Okawa, H.Oka, H.Yamada, S.Harihara, *T.Kuroki, *K.Kobayashi. Department of Gastroenterology, Osaka City General Hospital and "Third Department of Internal Medicine, Osaka City University Medical School, Osaka, Japan There have been reports sug~esling that interferon(IFN) affects lipid metabolism in animals. However, despite extensive clinical use of IFN in treatment of chronic hepatitis and malignant tumors, changes in lipid metabolism associated with IFN administration to humans have been only minimally studied. In addition, no comparison has been made of the effects of IFN alpha( a ) and IFN beta(/3 ) on lipid metabolism. In this study, 44 patients with chronic hepatitis C were divided into a group treated with IFN a ( a group, 22 patients) and one treated with IFN /3 (/3 group, 22 patients), and changes in lipid metabolism during IFN therapy were evaluated. In the c~ group, the serum free fatty acid level (FFA) increased significantly (p<0.01.) from 0.29±0.16 mEq/l before IFN administration to 0 . 4 2 + 0 . 2 3 mEq/1. during administration. In the fl group, sis0, it increased significanfly (p<0.01) from 0.35 ± 0.18 mEq/1 before IFN administration to 0.55 ± 0.34 mEq/1 during administration. I n t h e a group, the serum triglyceride level (TG) increased significantly (p<0.01) from 110.0 ± 52.3 mg/dl before IFN administration to' 130.5 ± 48.1 mg/dl during administration. In the/3 group, also, it increased significantly (p<0.01) from 127.0 ± 68.7 mg/dl before administration to 298.4 ± 267.7 mg/dI during administration. However, no significant changes were observed in serum total cholesterol level (T-CHO) or serum glucose level during IFN administration in either group. Conclusions. These fIndings suggest that IFN administration increases serum TG and FFA in patients with chronic hepatitis C. No difference in the effects on I!pid metabolism was observed between the a group and fl group. Since IFN administration markedly increase serum TG in some patients, and hypertriglyceridemia of more than 1000mg/dl is associated with a high risk of pancreatitis, careful observation of patients receiving IFN will be required for increase in serum TG and other side effects.
SERUM LEVELS OF TRANSFORMING GROWTH FACTOR ALPHA IN PATIENTS WITH CHRONIC HEPATITIS C. S.Kira,T.Nakanishi,R.Nakashio,M.Kitamoto,Y.Watanabe, G.Kajiyama,S.Suemori*.First Dept. of Internal Medicine,Hiroshima University School of Medicine,Hiroshima,Japan.*Gastrointestinal Unit,Yokohama City Seibu Hospital,St.Marianna University School of Medicine, Yokohama,Japan.
Previous studies have indicated that transforming growth factor alpha (TGF-alpha) play an important role in liver regeneration and hepatocarcinogenesis.We also have reported that serum TGF-alpba levels was el~va~l in patients with chronic hepatitis and hepatocellular carcinoma.Aim:The aim of this study was to assess the changes of serum TGF-alpha levels and efficacy of interferon (IFN) therapy in patients with chronic hepatitis C. Materials and Methods:Using enzyme-linked immunosorhent assay kit (Oncogene S~ienee Co.),we studied the serum 'levels of TGF-alpha (before,at the end of and 6 months after IFN therapy) in 14 patients with chronic hepatitis C who had been pathologically diagnosed as chronic aggresive hepatitis(CAH)2a by aspiration biopsy and had undergone IFN therapy.Seven patients with complete response had maintained normal ALT value for over 6 months after IFN therapy.And seven patients with non response had maintained abnormal high ALT value during the follow up. Results: In non response cases, the mean (+SD) TGF-alpha levels was 283.27 +32.27pg/ml before therapy,296.81+--29.64 pg/ml at the end of therapy and 284.87+42.36 pg/ml 6 months after therapy.In contrast,in Complete response cases,the mean (+SD) TGF-alpha levels was 209.80 +68.86 pg/ml before therapy,152.28+ 65.16 pg/ml at the end of therapy and 117.32+ 68.12 pg/ml 6 months after therapy.in complete response cases,serum TGF-a!pha levels at the conclusion of and 6 months after IFN therapy were lower than that before IFN therapy(P<0.05).There was significant statistical correlation between serum TGF-alpha le~,els and Serum ALT values. Conclusion:These results Suggest that serum TGF alpha levels may reflect the degree of liver damage and TGF-alpha may play a role in liver regeneration.
GASTROENTEROLOGY,Vol. 108, No. 4
COEXPRESSION OF TRANSFORMING GROWTH FACTOR ALPHA AND EPIDERMAL GROWTH FACTOR RECEPTOR IN HUMAN HEPATOCELLULAR CARCINOMA. S,Kira,R.Nakashio,M.Kitamoto,Y.Watanabe,T.Nakanishi, G.Kajiyama. First Dept. of Internal Medicine,Hiroshima University School of Medicine,Hiroshima,Japan. Previous studies have indicated that transforming growth factor alpha (TGF-alpha) acts on epidermal growth factor receptor (EGFR) and play an important role in liver regeneration and hepatocarcinogenesis.We aslo have reported the elevation of serum TGF-alpha levels in patients with hepatocellular carcinoma(HCC) by enzyme-linked immunosorbent assay. Aim: To compare the immunoaclfivity with the proliferation and the degree of historogic differentiation of HCC,the coexpression of TGF-alpha and EGFR was examined in the same area of HCC by immunohistochemical studies.Materials and Methods: Specimens of 48 nodules of HCC less than 5cm in diameter(17:well differentiated,25:moderately differentiated, 6:poorly diffe~rentiated ) were obtained from 44 Japanese patients who had undergone surgical resection.Immunoreactiv e TGF-alpha and EGFR in the same area of paraffin-embedded sections were detected by using monoclonal antibody(Oneogene Science Co.,Bio Genex Co.). Results:In adjacent non tumorous tissue,TGF-alpha was weakly expressed in all cases and EGFR was expresse'd in only 10 of 48(20.8%)cases.The expression of TGF-a!pha and EGFR in HCC was as follows; " TGF-alpha EGFR well (17) moderately (25) poorly (6) total (÷) • (+) 6/17 7/25 1/6 (14) (+) (-)' 7/17 7•25 1/6 (15) (+) (+) 0/17 3/25 4/6 ( 7) ('4") (:) 4/17 8'/25 0/6 (12) ( + )~: strongly 15ositive,(+) : weakly positive, (-) : negative In the cases Showing strongly positive staining of TGF-alpha,EGFR was expressed at almost equal frequency.Otherwise,in the cases showing weakly positive staining of TGF-alpha,the expression of EGFR differs statistically ~ccording to the degrees of histologic differentiation of HCC(p<0.05).In comparison with well or morerately differenciated HCC,poorly differentiated HCC expresses EGFR highly with the enhaficed proliferative responce to weak TGF-alpha expression.Regarding the tumor diameter and cumulative recurrence rates,significant difference was found between EGFR-positive groups and EGFR-negative groups(p<0.05). Conclusion: These results suggest that TGF-alpha / EGFR may play a role in the differentiation and the proliferation of HCC
TRANSFORMING GROWTH FACTOR ALPHA ANTISENSE DNA OLIGONUCLEOTIDES APPROACH TO INHIBIT GROWTH OF HUMAN HEPATOCELLULAR CARCINOMA DERIVED CELL LINES. S.Kira,T.Nakanishi,R.Nakashio,M.Kitamoto,Y.Watanabe, G.Kajiyama,S.Suemori*.First Dept. of Internal Medicine,Hiroshima University School of Medicine'Himshima'Japan'*Gastr°intestinal Unit,Yokohama City Seibu Hospital,St.Marianna University School of Medicine, Yokohama,Japan.
Previous studies have indicated that transforming growth factor alpha (TGF-alpha) play an important role in liver regeneration and hepatocarcinogenesis.We also have reported that serum TGF-alpha levels was elevated in patients with chronic hepatitis and hepatocellular carcinoma(HCC). Aim:To better understand the role of TGF-alpha in development of HCC,we investigated the specific inhibition of TGF-alpha gene by TGF-alpha DNA ann-sense posphorothioate oligonucleotides (SODN,Oncogene Science Co.) in HCC derived cell lines. Materials and Methods: 1) Human HCC derived cells (Hep G2,HLF)were plated at 1.0xl04cells/well in 96 well plates and grown in 100gl of WEM supplemented with 10% FCS.Human TGF-alpha SODN and control oligonucleotides were added directly into the medium 48hrs after plating cells.3H thymidine incorporauon was measured 24hrs after addition.2)TGF-alpha in culture medium was measured by using enzyme-linked immunosorbant assay kit (Oncogene Science Co.). Results: TGF-alpha in culture medium was 6.71 _l.57pg/24hr/g protein (HepG2), 4.99 +-1.58pg/24hr/g protein (HLF).3H thymidine incorporation ratio to non addition controls was as follows; TGF-alpha S-ODN Control oligonucleotides 0.5 gtM 1.01.tM 2.0p.M 0.5 ~tM 1.0gM 2.0p.M HepG2 96% 76% 72% 1 0 8 % 106% 94% HLF 96% 82% 89% 125% 110% 140% Conclusion:These results suggest that TGF-alpha could be an autocrine factor in hepatocellular carcinoma derived cell lines and TGFalpha may play a role in development of hepatocellular carcinoma.