Ouabain에 의한 心臟障害에서 心臟組織의 ATPase 및 RNase活性의 變動에 대한 硏究

李正浩,金近浩,高在炅 한양대학교 의과대학 1982 한양의대 학술지 Vol.2 No.1

In order to elucidate the nature of cardiac toxicity induced by ouabain, activities of adenosine triphosphatases(ATPases) in heart and serum from rats treated with ouabain were determined and they were compared with those from normal rats. Also determined were activities of acid and alkaline RNases from heart and serum from the rats using RNA, poly A or poly C as substrates. The results obtained were as follows: 1) While the activity of ??-ATPase in heart homogenate was influenced by neither ??,??,nor ouabain, a distinct activity for ??,??-ATPase was observed in both the heart cell fractions of 10⁴×g supernatant and precipitate. 2) Activities of ??,??-ATPase in both mitochondrial and microsomal fractions of heart tissues were significanlty decreased in ouabain treated rats as compared with those of control rats. While cardiac ?? content was significantly increased, ?? content was significantly decreased. 3) In in vitro experiments the activity of cardiac ??-ATPase not influenced by ouabain added to the assay medium, but ??-ATPase activities in heart homogenate and the cell fractions were decreased in rats treated with ouabain as compared with those in control rats. These results indicate that the administration of ouabain leads to changes in myocardial ?? and ?? contents by inhibiting mitochondrial and microaomal ?? and ?? contents by inhibiting mitochondrial and microsomal ??,??-ATPase activities, which may in turn, rusult in the disturbance of myocatdial contractility to induce cardiac toxicity. It can not be ruled out, however, the possibility that a decrease in cardiac ??-ATPase activity may be correlated, in part,with cardiotoxicity induced by ouabain. 4) Activities of cardiac acid RNase was significantly decreased when measured using Rna as the substrate and was increased using poly C as the substrate in rats treated with ouabain as compared with those in control rats. On the other hand, activities of cardiac alkaline RNase was significantly decreased when measured using RNA or poly A as the substrate. These results suggest that the administration of ouabain leads to damages in structural integrity of myocardium. 5) The mean values of activities of serum ATPases and RNases appeared to be higher in rats treated with ouabain than in control rats, but exhibited no statistical significance. The results mean that in ouabain cardiotoxicity the cardiac tissue enzymes may not be released into the blood stream or the amount of tissue enzymes released into the blood may not be large enough for giveing rise significant changes in activities of the serum enzymes.

암의 생화학적지표로서의 Acid Deoxyribonuclease에 대한 연구

박장수,김용석,고재경 한양대학교 의과대학 1995 한양의대 학술지 Vol.15 No.1

Nucleic acid content, protein content and acid DNase activity were determined in human cancer tissues from lung, digestive, urological, gynecological and bone systems. The positive rate of the acid DNase as a biochemical marker for cancer was also determined. The acid DNases were partially purified from a part of human cancer tissues studied and were subjected to study for substrate specificity to understand the possible role of the enzyme in carcinogenesis. The pattern of changes in concentrations of nucleic acid and protein in sixteen human cancer tissues studied appeared to be variable in nature. Increases in both nucleic acid and protein contents were observed in tumor tissues of stomach cancer, urinary bladder cancer, renal cell carcinoma, uterine cervix cancer, osteosarcoma and metastatic squamous cell carcinoma, increases in nucleic acid contents without changing protein contents were seen in tumor tissues of four types of lung cancer, hepatocellular carcinoma and serous cystadenocarcinoma of ovary and increase in protein contents without changing nucleic acid content was found in giant cell tumor tissue of bone. Both nucleic acid and protein contents were unchanged in mucinous cystadenocarcinoma and endodermal sinus tumor tissues of ovary. These results indicated that changes in nucleic acid and protein contents could be used as a biochemical marker for certain cancer. Activity of acid DNase known to be involved in DNA integration and carcinogenesis was greatly increased in 14 human cancer tissues out of 16 tumor tissues studied. The positive rate of the acid DNase as markers for cancer was high in value suggesting the possible use of the enzyme as biochemical markers of these cancer. The acid DNases were partially purified by 7 to 63 folds in eight human cancer tissues(epidermoid carcinoma of lung, stomach cancer, urinary bladder cancer, uterine cervical cancer, serous cystadenocarcinoma and endodermal sinus tumor of ovary, osteosarcoma and metastatic squamous cell carcinoma of bone) and isolated to be a single enzyme, not isozyme forms. The acid DNase purified was highly acitve toward double stranded (ds) DNA in gereral. Low or no acitivties toward single stranded (ss) DNA, RNA and polyribonucleotides were observed. Specificity toward ds DNA was variable among the acid DNase purified from cancer tissues studied. The specificity toward ds DNA was the highest in acid DNase from uterine cervical cancer tissue (3%), lower in the enzyme from serous cystadenocarcinoma of ovary, urinary bladder cancer, metastatic squamous cell carcinoma of bone and stomach cancer tissues(82-88%) and the lowest in the enzymes from epidermoid carcinoma of lung, osteosarcoma and endodermal sinus tumor of ovary tissues. The assumption that the acid DNase might play a role in transforming normal cells into cancer cells by integrating foreign DNA into host chromosomal DNA or by modifying a part of host genome might be suggestive in the development of cervical cancer of uterus, but not of other tumor tissues studied.

간 혼수를 동반한 간세포암종 환자의 뇌척수액 Ribonuclease 기질특이성에 관한 연구

변동일,김동권,이용성,고재경 한양대학교 의과대학 1996 한양의대 학술지 Vol.16 No.1

Activity of ribonuclease (RNase) was determined with olycytidylate (poly C) and RNA as substrate in the cerebrospinal fluid(CSF) of patients with hepatocellular Carcinoma with coma. RNase and proteins in the CSF of patients with hepatocellular carcinoma were isolated and purified y a DEAE-cellulose column chromatography and high perormance liquid chromatography (HPLC). Substate specificity for the purified RNase activated in the CSF of the cancer patient was determined to evaluate the possible role of the RNase in car5cinogenesis. Activity of RNA measured with poly C and RNA as substrate and ratio of RNA/poly C for the RNase activity in the csf were increased significantly in the hepatocellular carcinoma, but were unchanged in the liver cirrhosis. The positive rate of the RNAse activity in the CSF as a marker for hepatocellular carcinoma was high, suggesting the use of the RNase activity in the CSF as a marker for hepatocellular carcinoma. Proteins and RNases in the CSF of the patient with hepatocellular carcinoma were separated into 7 protein peaks and 5 RNase isozymes(isozyme Ⅰ, Ⅳ-Ⅶ), of which two peak proteins and one RNase isozyme(isozyme Ⅰ) were specific to the hepatocellular carcinoma. The Rnase isozyme Ⅰ purified from the CSF of the cancer patient was a major isozyme of the RNase isozymes isolated, was activated and was higher in ratio of RNA/poly C for the RNase activity, exhibiting more of nonsecretory RNase nature than that from the CSF of the control. The RNase isozyme Ⅰ fraction in the Csf of the patient with hepatorcellular carcinoma was separated by a HPLC into four subpeaks, of which cubpeak Ⅰ-3 exhibited RNase activity. The RNase isozyme Ⅰ (Ⅰ-3) purified from the CSF of the patient with hepatocellular carcinoma showed the highest activity toward poly AC as substrate, the degree of activity being decreased toward poly ACU and C, AU, CI, CIU, CU, RNA, poly U in order. No activity was observed with poly A, G, AG and GU. The substrate specificity toward the RNAse isozyme Ⅰ from the CSF of the cancer patient appeared to be different from that of the control in that relative activity toward RNA, poly U, CI, AC, AU, CIU aand ACU over poly C was higher in the isozyme from the CSF of the cancer patient than in the isozyme from the CSF of the control. The result indicated that the RNASE isozyme Ⅰ from the CSF of the cancer patient was different in nature from the isozyme from the CSF of the control. Observations that in the CSF of the patient with hepatocellular carcinoma with hepatic coma, (1) five RNase isozymes were isolated, (2) of which one isozyme (isozyme Ⅳ) was specific to hepatocellular carcinoma, (3) the RNase isoyzme Ⅰ isolated as a major enzyme was activated and (4) the substrate specficity of the isozyme in the CSF of the cancer patient was different from that in the CSF of the control, suggested that the RNase isozyme Ⅰ in the CSF of the cancer patient might be involved in carcinogenesis processes of hepatocellular carcinoma.

림프구성백혈병 환아의 혈청에서 RNase작용의 조절에 관한 연구

이철호,정성철,이상훈,고재경 한양대학교 의과대학 1996 한양의대 학술지 Vol.16 No.1

Ribonuclease(RNase) inhibitor has been known to specifically inhibit RNase known to be involved in carcinogenesis and suppression of cancer. In order to understand interactions between RNase and Rnase inhibitor in serum of patients with acute lymphocytic leukemia (ALL), the serum was pretreated with parahydroxymer-curibenzoate(PHMB) and then was chromatographed to remove RNAse inhibitor from RNase-RNase inhibitor comples. Properties of the inhibitor released RNAse thus obtained from the ALL serum pretreated with PHMB were compared with those from the ALL serum without PHMB pretreatment and with those from the control serum. Rnases in the AKK serum without PHMB pretreatment were separated by a DEAR-cellulose column chromatography into five isozymes (free RNase isozyme Ⅰ-Ⅴ), of which three isozyme activities were greater than those in the control serum, suggesting that free RNase isozymes Ⅰ,Ⅱ and ALL serum were activated. RNase inhibitor activity was higher in all of the four free RNase isozymes (Ⅱ-Ⅴ) except for the isozyme Ⅰ, being highest in the isozymeⅤ. RNases in the ALL serum tiwh PNMB pretreatment were also separtated by the chromatography into five isozymes (inhibitor released RNase isozyme Ⅰ-Ⅴ), of which the inhibitor released RNase isozyme Ⅰ activity being markedly higher than free RNase ixozyme Ⅰ activity from ALL serum without PHMB pretreatment. The RNAse inhibitor acitivity was greatly reduced in each of the isozymes isolated from the serum with PHMB pretreamtent. The results indicated that RNases in the ALL serum pretreated with PHMB were present in the state of inhibitor released RNase isozyme, accumulating in the isozyme Ⅰ. Activities of all of the free RNase isozymes separated from the ALL serum except for the isozyme Ⅴ were higher toward poly C than toward RNA as substratc, ratio of RNA/poly C being lower, suggesting the nature of secretory type of RNase, Ratio of RNA/poly C in the isozyme Ⅴ from the ALL serum was higher than that from the control serum and was close to 1.0 suggesting the nature of nonsecretory type of RNase. The results indicated that the free RNase isozyme Ⅴ isolated from the ALL serum was different from that from the control serum. Ratio of RNA/poly C of each of the inhibitor released RNase isozyme activity from the ALL serum with PHMB pretreatment was similar to each of those from the control serum, except for the isozyme Ⅰ. Ratio of RNA/poly C of inhbitor released RNase isozymeⅠ activity from the ALL serum was greater than that of inhibitor released RNASE isozyme Ⅰ activity from the control serum and that of free RNase ixozyme Ⅰ activity from the ALL serum. These results suggested that the inhibitor released RNase isozymes isolated from the ALL serum were different in nature from the free RNase.

위선암 환자의 위암조직과 위액의 RNase Isozyme, RNase Inhibitor 및 단백의 상호관계에 관한 연구

김기형,이준규,이상훈,고재경 한양대학교 의과대학 1996 한양의대 학술지 Vol.16 No.1

Ribonuclease (RNase) known to be associated with carcinogenesis processes was determined in its activity in stomach cancer tissue and gastric juice of patients with stomach cnacer. Rnases, Rnase inhibitoras and proteins were isolated from the cancer tissue were studied. The results were compared with those obtaind from gastric juice and stomach tissue of normal control in order to find out whether cancer specific RNases were present and were released from the cancer tissue into the gastric juice. Rnases wer present and were released from the cancer tissue into the gastric juice. Rhase activity was significantly decreased and RNase inhibitor activity was unchanged in the stomach cancer tissue, while RNase and Rnase inhibitor activities were markedly increased in gastric juice of patients with stomach cancer. This indicates that Rnase activities in the gastric juice could be used as biochemical and clinical markers for the stomach cancer. DEAE-cellulose oclumn chromatography revealed that proteins and RNases in the stomach cancer tissue were separated into 7 protein peaks and 6 isozymes each, of which one Rnase isozyme appeared to be specific to the cancer. Proteins and RNases in the gastric juice of the patients with stomach cancer were separated into 8 protein peaksand 4 isozymes each, of which 2 proteins and 4 isoyzmes were speicfic to the cancer. Substrated specificity, ratio of Rnase inhibitor/Rnase and ratio of RNA/poly C for RNase activity of RNase isozymes isolated from the gastric juice of patients with stomach cancer were observed to be similar to those of RNase isozymes isolated from the stomach cancer tissue, indicating that the Rnase isozymes in the gastric juice were released from the cancer tissue.

춘계학술대회 : 백반증 심포지움 ; 백반증의 병리조직학적 변화

고재경 대한피부과학회 1995 대한피부과학회 학술발표대회집 Vol.47 No.1

지역의 탄소중립,목표 선언 넘어 실행이 중요!

고재경,예민지 경기연구원 2021 이슈&진단 Vol.- No.451

난소암 환자 혈청 Amylase의 분리와 성상에 대한 연구

정성노,김문신,김두상,고재경 한양대학교 의과대학 1983 한양의대 학술지 Vol.3 No.2

In order to observe changes in serum amylase activities in various ovarian diseases including ovarian cancers, activities of serum amylase were measured in patients with functional cyst, cystadenoma and cystadenocarcinoma and were compared with those of normal controls. Serum amylases were separated and analyzed by DEAE-cellulose chromatography and by electrophoresis on cellulose acetate membrane to see whether an amylase specific to the ovarian cancer was present in serum of patients with serous cystadenoma. 1) Activities of serum amylase were significantly increased in serous cystadenoma, mucinous cystadenocarcinoma, serous cystadenocarcinoma and Krukenberg tumor among the ovarian diseases studied, the highest activity being observed in serous cystadenocarcinoma. 2) DEAE-cellulose column chromatography of serum for amylase showed that two types (peak Ⅰ and Ⅱ) of amylase or group of amylases were present in serum of both normal controls and patients with serous cystadenocarcinoma and that the peak Ⅰ amylase activity was markedly greater in serous cystadenocarcinoma than in normal control. 3) The properties of the peak Ⅱ amylase in serous cystadenocarcinoma might be different from those in normal control, since the peak Ⅱ enzyme in both the ovarian cancer and normal control showed different patterns of substrate specificity. 4) Protein patterns of the peak Ⅰ and Ⅱ fractions obtained by electrophoresis were shown to be different between serous cystadenocarcinoma and normal control: A protein band 2 specific for the ovarian cancer was observed in the peak Ⅰ fraction of serous cystadenocarcinoma and protein bands 2 and 3 in the peak Ⅱ fraction were greater in the ovarian cancer than in normal control. The present sutdy indicated that an amylase specific for the ovarian cancer might be present in serum of patients with serous cystadenocarcinoma and that the enzyme could be useful as a marker for the ovarian cancer.

난소암 환자의 복수 효소활성 및 단백분획의 변동에 대한 연구

권혜숙,고재경 한양대학교 의과대학 1984 한양의대 학술지 Vol.4 No.2

In order to observe changes in enzyme activities in ascitic fluids of ascites-forming ovarian diseases, activities of nucleases, 5'-nucleotidase and amylase in ascitic fluids from patients with non-tumorous ovarian cysts, Krukenberg's tumor, cystadenoma and cystadenocarcinoma were determined. Alsostudied were changes in protein patterns of ascitic fluids from patients with the ovarian diseases. 1) While activities of ribonuclease (RNase) in ascitic fluid were unchanged in Krukenberg's tumor, cystadenoma and mucinous cyst adenocarcinoma as compared with that of simple ovarian cyst, the activity was increased significantly by 20% in serous cystadenocarcinoma. 2) Activities of 5'-nucleotidase in ascitic fluid were unchanged in Krukenberg's tumor. Activities of 5'-nucleotidase in ascitic fluid were, however, increased significantly in cystadenoma and cystadenocarcinoma. 3) Amylase activities in ascitic fluid were unchanged in Krukenberg's tumor and mucinous cystadenoma. On the other hand, amylase activities in ascitic fluid were significantly increased in serous cystadenoma and cystadenocarcinoma, the greatest increment (6.5-fold increase) being observed in serous cystadenocarcinoma. 4)Poteins in ascitic fluid were ciearlyseparated into five bands on a cellulose acetate membrane by electrophoresis. In serous cystadenocarcinoma protein content of the band 1 was significantly decreased and that of the band 3 increased. The results obtained in the present study suggested that increases in enzyme activities and changes in protein patterns of ascitic fluid from patients with serous cystadenocarcinoma might be, in part, due to alterations in nucleic acid and protein metabolism observed in ovarian cancer tissues and body fluids.

胃癌組織內의 Acid Deoxyribonuclease 活性 및 性狀에 關한 硏究

咸駿洙,朴炅南,高在炅 한양대학교 의과대학 1985 한양의대 학술지 Vol.5 No.1

In the present study, activities of enzymes involved in deoxyribonucleic acid (DNA) degradation, deoxyribonuclease (DNase) Ⅰ, Ⅱ, Ⅲ and Ⅳ, were measured in extracts of stomach cancer tissues (20 cases) and were compared with those in extracts of normal control tissues. In order to clarify a possible association of acid DNase with carcinogenesis of stomach cancer, characteristic properties of acid DNase and mechanism of hydrolytic cleavage of DNA by acid DNase in stomach cancer tissues were studied. 1. Contents of DNA and protein were significantly higher in stomach cancer tissues than in normal control tissues, but that of RNA was not changed. 2. Of 4 types of DNase studied, the highest activity was observed with acid DNase (DNase Ⅱ) in both normal control and stomach cancer tissues, and the activity of acid DNase was significantly higher in stomach cancer tissues than in normal control tissues. 3. Acid DNase in stomach cancer tissues hydrolyzed double stranded DNA more rapidly than did single stranded DNA, but did not exhibit absolute specificity toward double stranded DNA. 4. Most of the products of enzymatic hydrolysis of DNA by acid DNsase in stomach cancer tissues were found to be polydeoxyribonucleotides with nucleotide length more than 20. Some of the hydrolytic products were, however, observed to be monodeoxyribonucleotides. Observations that the activity of acid DNase in stomach cancer tissues 1) was significantly increased, 2) was highly active against double stranded DNA and 3) cleaved DNA endonucleolytically support the assumption that the DNase might play a role in transforming normal cells into cancer cells by modifying a part of host genome.