15d-PGJ2 inhibits NF-kB and AP-1-mediated MMP-9 expression and invasion of breast cancer cell by means of a heme oxygenase-1-dependent mechanism

장혜연,On-Yu Hong,Hyun Jo Youn,김민걸,Cheorl-Ho Kim,정성후,Jong-Suk Kim 생화학분자생물학회 2020 BMB Reports Vol.53 No.4

Activation of peroxisome proliferator-activated receptor  (PPAR) serves as a key factor in the proliferation and invasion of breast cancer cells and is a potential therapeutic target for breast cancer. However, the mechanisms underlying this effect remain largely unknown. Heme oxygenase-1 (HO-1) is induced and overexpressed in various cancers and is associated with features of tumor aggressiveness. Recent studies have shown that HO-1 is a major downstream target of PPAR. In this study, we investigated the effects of induction of HO-1 by PPAR on TPAinduced MMP-9 expression and cell invasion using MCF-7 breast cancer cells. TPA treatment increased NF-B /AP-1 DNA binding as well as MMP-9 expression. These effects were significantly blocked by 15d-PGJ2, a natural PPAR ligand. 15d-PGJ2 induced HO-1 expression in a dose-dependent manner. Interestingly, HO-1 siRNA significantly attenuated the inhibition of TPA-induced MMP-9 protein expression and cell invasion by 15d-PGJ2. These results suggest that 15d-PGJ2 inhibits TPA-induced MMP- 9 expression and invasion of MCF-7 cells by means of a heme oxygenase-1-dependent mechanism. Therefore, PPAR/HO-1 signaling- pathway inhibition may be beneficial for prevention and treatment of breast cancer.

Glycosyltransferases in human liverdiseases and cancer, and effects of oriental medicine on human HBV

Kim, Cheorl-Ho 경희대학교 동서의학연구소 1999 INTERNATIONAL SYMPOSIUM ON EAST-WEST MEDICINE Vol.1999 No.1

Cheorl-Ho Kim. Department of Biochemsitry and Molecular Biology, College of Oriental Medicine, DongGuk University,Kyungju, Korea Glycosyltransferases in human liver diseases and cancer, and effects of oriental medicine on human HBV. Proceedings of International Symposium on East-West Medicine, Seoul. 128-148,1999. -N-Acetylglucosaminyltransferase-Ⅲ(GnT-Ⅲ) which catalyzes the addition of NacGlc through αβ1-4 linkage to to the β-linked mannose of the trimannosyl core, seems to paly a key role in the alteration of N-glycan structures. We have used the transgenic mouse system to target the expression of human GnT-Ⅲ to the liver of the mouse. The GnT-Ⅲ transgene was designed to be expressed in the transgenic animal directed by the mouse albumin enhancer' promoter and SV40 polyA addition signal. Total 5 transgenic founder mice were generated by pronuclear microinjection and in 4 cases transformation of the transgenes was confirmed in the G1 generation. Northern blot analysis of total RNAs from various organs of the transgenic mice revealed that the expression of the human GnT-Ⅲ was specific to the liver. Upon western analysis the human GnT- Ⅲ protein was deteted only in the liver of transgenic mice, Moreover, these misc showed a gross abnormality in the abdomen, I,e., an abnoraml inflation and had a swollen oval-like morphology, with many lipid Droplets. We demonstrated with these results that aberrant glycosylation leads to the generation of fatty liver and suggest novel mechanism for the pathogenesis of fatty liver. Also, we have Estblished 6 hybridomas which produce anti-GnT-Ⅲ monoclonal antibodies, and applied these for the simple diagnosis of liver diseases by sandwich-ELISA method using the monoclonals. The application to diagnosis will be also discussed. Human hepatitis B virus(HBV) envelopes contain three distinct glycoproteins called L, M. and S HbsAg. There is uncertianty about what role, if any, these N-glycans and their modification in the ER play in the function of the HBV envelope proteins. By treating hepatoblastoma cultures which secrete HBV(Hep3b cells) with inhibitors of different steps of the glycosylation and glycan modifying pathway, we provide evidence that glycosylation and the first step in the processing pathway are necessary for virion, but not subviral particle secretion. That is, using a highly sensitive immunoprecipitation/PCR system, enveloped HBV could not be detected by tunicamycin. Moreover, inhibitors of α-glucosidase I (the first step in the glycan processing pathway)also preventede virion secretion. Inhibitors of mannose trimming (a later step) and glycolipid synthesis did not revent wirion secretion, defining the limits of the glycosylation requirements in secretion These results demonstrate a requirement for N-glycosylation and glucosidase processing in the secretion of virions and further distnguish between the requirements for virion and aubviral particle secretion. To investigate whether or not structural cnanges of oligosaccharides affect secretion and gene expression of HBV, we introduced the GnT-Ⅲ gene into a human hepatoma cell line, Hep3B, which secreted HBV-related proteins into the medium. The clones have high GnT-Ⅲ activities and secreted lower levels of HBV-related proteins into the medium. Also TOM_treated cell lines with GnT-Ⅲ transfectant showed low levels of secretion of HBV-related proteins into the medium. Thus, TOM was highly effective for liver hapatocarcinoma treatment and it was concluded that the medicine can be used as potent anti-liver cancer medicine.

枇杷葉의 抗糖尿病藥理活性과 IL-1β誘發糖尿病 마우스의 膵臟 燐酸化酵素에 미치는 效果에 關한 硏究

윤철호,정지천,김철호 동국대학교 한의학연구소 1998 東國韓醫學硏究所論文集 Vol.7 No.1

淸凉生津, 解渴上嘔의 효능이 있으며, 한국등에서 항당뇨병성 한약으로서 사용되어 온 비파엽의 열수출물의 당뇨병치료효과를 검증하기 위하여 alloxan으로 유발된 당뇨병흰쥐의 혈장포도당농도 및 인슈린농도를 측정하였다. 비파엽처리군의 혈장 포도당농도는 현저히 증가하였으며 혈장인슈린농도는 크게 감소하지 않았다. 한편, 당뇨병유발의 내재적인 요소로 알려진 사이토카안인 IL-1β로 당뇨병을 유발한 뒤 그 효과를 검정했다. 즉, 실험동물을 interleukin-1β로 처리하여 실험적으로 당뇨병을 유발한 후 glucose의 燐酸化에 관여하는 glutokinase와 hexokinase의 활성을 살펴보았다. 그 결과, 비파엽엑기스는 interleukin-1β 과잉투여 당뇨병으로 인하여 증가된 혈액내 glucose농도를 감소시키고 insulin 과잉분비를 정상으로 회복시키는데 효과적이었다. 동시에 lL-1β에 의해 유도된 당뇨마우스에 대한 枇杷葉엑기스 投與가 췌장 glucokinase와 hexokinase의 활성을 촉진하는 것으로 나타나 lL-1β유발 당뇨병에 효과가 있는 것으로 사료되었다. Studies were conducted on anti-dibetic effect of the water extract from leaves of Eriobotryae folium which had been used in Korea as a remedy for dibetes. The extract was found to inhibit the Increase in the plasma level of sugar bu the not the decrease in the plasma level of insulin in alloxan-induced dibetic rata. Also, we investigated the in vivo effect of an aqueous extract(referred to as EF) from Eriobotryae folium on glucokinase and hexokinase activities of diabetes mellilus induced by Interteukin-1β (IL-1β). After 1 week of IL-1β injection, the levels of serum glucose concentration and insulin secretion were dramatically increased. However, the insulin secretion was decreased with administration of EF. The level of glucose concentration was decreased by EF administration. Furthermore, it was observed that EF was effective in recovering the levels of insulin secretion. Enzyme activities of the glucokinase and hexokinase, which are key enzymes of glucose phosphorylastion, were decreased by IL-1β. EF administration to the mice allowed proportional increasing by stimulation of induction of enzyme activities as high as normal group. These results suggested that EF is highly effective in treatment of diabetes mellitus induce by IL-1β.

비지의 발효과정 중 기능성 식품효소를 생산하는 미생물의 분리 및 특성

김동수,김철호 慶星大學校 1998 論文集 Vol.19 No.2

This study was attempted to isolate and characterize microorganisms in fermenting soybean curd residues. The isolated microorganisms were mainly belonged to the genus of Bacillus species and fugal Aspergillus species. The strains were cultivated for production of carbohydrate-related enzymes and proteolytic enzymes such as β-fructofuranosidase, pullulanases, amylases, galactosidases, and proteases. These enzymes are useful in development of food resources like artificial food containing galactooligosaccharides, fructooligosaccharides or nondigestive-oligosaccharides. Among them, one β-fructofuranosidase was produced from a Bacillus strain and purified by ammonium sulfate gractionation, DEAE-sephacel chromatography and gel filtration of Sepharose-6B (FPLC) chromatography from two days-grown bacterial cells. Molecular weight of the purified enzyme was estimated to be about 56,000 (Mr). Now we are examining the properties and characterization of the bacterium and enzyme and large-scale production of useful oligosaccharides for food resources.

五倍子가 Alloxan에 의한 實驗的 糖尿病 마우스의 膵臟 glucokinase 및 hexokinase活性에 미치는 影響에 關한 硏究

정지천,김철호,최형일 東國大學校 1998 東國論叢 Vol.37 No.-

Glucokinase와 hexokinase는 간과 췌장에서 인슐린에 의해 혈당이용을 증대시키며 혈당농도를 감지하여 인슐린 분비를 조절하는 효소로서 인슐린비의존성당뇨병(NIDDM)의 병인과 밀접한 관계가 있는 것으로 알려져 있다. 또한, alloxan은 glucokinase의 활성부위에 존재하는 -SH기와 경쟁적으로 결합하여 효소활성을 저해함으로서 당뇨병을 유도한다. 본 연구에서는 당뇨병치료의 전통한방약재로서 사용되고 있는 五倍子추출물의 효과를 해명하기 위하여 alloxan유발 실험적 당뇨마우스의 glucokinase와 hexokinase의 활성을 검토한 겨과, alloxan주사에 의해 체중과 공복시의 glucose 및 insulin 분비가 증가되었으나, 五倍子투여로 공복시의 insulin분비가 감소하였으며 체중과 공복시의 혈당은 유의성은 없었으나 대조군에 비하여 감소하였다. 그러나, 혈당수치가 정상군에 비하여 현저하게 증가되었으나 五倍子투여로 현저히 감소하였으며 Insulin치의 상승과 분비지연도 五倍子투여군에서는 정상군과 유사한 경향을 보였다. 한편, 혈중glucokinase와 hexokinase의 활성은 alloxan주사에 의해 현저히 감소되었으나, 五倍子 투여로 유의성 있게 상승되었다. 이상의 결과로, 五倍子는 alloxan으로 유발된 실험적 당뇨병에서 포도당인 산화 효소인 glucokinase와 hexokinase의 활성을 증가시키는 것으로 나타나 당뇨병의 치료에 임상적인 효과가 있음을 시사하였다. The in vivo effect of an aquous extract from Rhois Galla (R-G) on glucokinase and hexokinase activities has been examined in mice having diabetes mellitus induced by alloxan, which specifically bind to -SH group of the enzymes and inhibits the enzyme activities. After 1 week of alloxan injection, the levels of serum glucose and insulin secretion were dramatically increase, however, the insulin secretion was decreased with administration of R-G in alloxan-treated mice. Alloxan injection allowed the serum glucose level increased and the level was decreased by R-G administration. Furthermore, it has been observed that R-G was effective in recovering the levels of insulin secretion in the alloxan-treated diabetic mice. Enzyme activities of the glucokinase and hexokinase were decreased by alloxan treatment. In contrast, R-G administration to the mice allowed proportional increasing. These results strongly suggested that R-G is highly effective in treatment of diabetes mellitus induced by alloxan.

Cytokine interleukin-1β로 誘發된 糖尿病 마우스의 膵臟 glucokinase 및 hexokinase 活性에 對한 五倍子의 效果에 關한 硏究

최형일,정지천,김철호 동국대학교 한의학연구소 1998 東國韓醫學硏究所論文集 Vol.7 No.1

Glucokinase와 hexokinase는 간과 췌장에서 인슐린에 의해 혈당이용을 증대시키며 혈당농도를 감지하여 인슐린 분비를 조절하는 효소로서 인슐린비의존성당뇨병(NIDDM)의 병인과 밀접한 관계가 있는 효소이다. Cytokine의 일종인 interleukin-1β (IL-1β)는 췌장 도세포에서의 인슐린 분비와 β-cell에 대한 세포 독성을 효과적으로 조절하여 장기간 노출시키면 인슐린 분비와 생합성을 저해하며 β-cell을 파괴하여 β-cell의 기능적, 구조적 손상을 초래하여 IDDM을 유발한다. 본 연구에서는 당뇨병 치료의 전통한방약제로서 사용되고 있는 五倍子추출물의 효과를 해명하기 위하여 IL-1β유발 실험적 당뇨마우스의 glucokinase와 hexokinase의 활성을 검토한 결과, lL-1β주사에 의해 체중과 공복시의 glucose 및 insulin 분비가 증가되었으나, 五倍子투여로 공복시의 insulin분비가 감소하였으며 체중과 공복시의 혈당은 유의성은 없었으나 대조군에 비하여 감소하였다. 그러나, 혈당수치가 정상군에 비하여 현저하게 증가되었으나 五倍子투여로 현저히 감소하였으며 Insulin치의 상승과 분비지연도 五倍子투여군에서는 정상군과 유사한 경향을 보였다. 한편, 혈중 glucokinase와 hexokinase의 활성은 IL-1β주사에 의해 현저히 감소되었으나, 五倍子 투여로 유의성 있게 상승되었다. 이상의 결과로, 五倍子는 IL-1β로 유발된 실험적 당뇨병에서 포도당인산화 효소인 glucokinase와 hexokinase의 활성을 증가시키는 것으로 나타났다. We investigated the in vivo effect of an aquous extract from Rhois Galla (R-G) on glucokinase and hexokinase activities of diabetes mellitus induced by interleukin-β (IL-1β). After 1 week of alloxan injection, the levels of serum glucose and insulin secretion were dramatically increased, however, the insulin secretion was decreased with administration of R-G, IL-1β injection allowed the scrum glucose level increased and the level was decreased by R-G administration. Furthermore, we could observe that R-G was effective in recovering the levels of insulin secretion. Enzyme activities of the gtucokinase and hexokinase were decreased by IL-1β treatment. In contrast, R-G administration to the mice allowed proportion increasing. Seemingly, when IL-1β was injected to the mice, enzyme activities of the glucokinase and hexokinase were decreased. But, R-G stimulated induction of enzyme activities of the glucokinase and hexokinase as high as normal group. These results suggested that R-G is highly effective in treatment of diabetes mellitus.

黃柏에서 抽出한 多糖類의 抗癌作用에 대한 硏究

최영철,홍희탁,홍영욱,이태균,박원환,김철호 동국대학교 경주대학 1997 東國論集 Vol.16 No.1

In order to investigate antitumor activities of the extracted polysaccharides from Phellodendron chinense SCHNEID, the following studies were carried out. Phellodendron chinense SCHNEID was extracted with hot water and alkali, and the extracted polysaccharides showed antitumor activities against mice Sarcoma-180 solid tumors in parameters of tumor weight, inhibition ratio, thymidilate synthetase(TS) and thymidine kinase(TK) activities. The results were as follows : 1. Delipidated extracts from Phellodendron chinense SCHNEID, were systematically fractionated, the heteropolysaccharide contains D-N-acetylglucosamine, D-galactose, D-mannose, D-glucose. The molecular weight distribution of the sample was found to be nearly homogeneous (about from 200 to 1000 kDa) in gel permeation analysis. 2. Among the polysaccharide fractions, the water extracted fractions, particularly Fr.-2, showed a potent growth-inhibitory activity against implanted in mice Sarcoma 180 solid tumor (inhibition, 95.3%, at 2 ㎎/100g), but the cold alkali extract (Fr.-4) showed a very low activity (inhibition; 4.7%). Fr.-7 and Fr.-8 showed the highest tumor inhibitory activity (inhibition; 96.4 and 98.2%, respectively). 3. TS activity in the Fr.-2 given oral administration of propolis was markedly suppresed to 73.7% of that in the control (p<0.01 by Student's t-test). However, there was little difference in TK activity with a slight decrease in Fr.-2 only.

A Salmonella typhimurium rfaE Mutant Recovers Invasiveness for Human Epithelial Cells when Complemented by Wild Type rfaE

Cheorl-Ho Kim 한국분자세포생물학회 2003 Molecules and cells Vol.15 No.2

Glycoantigen Biosyntheses of Human Hepatoma and Colon Cancer Cells are Dependent on Different N-Acetylglucosaminyltransferase-3 and -5 Activities

( Cheorl Ho Kim ) 한국미생물생명공학회 2004 Journal of microbiology and biotechnology Vol.14 No.5

Human Hepatocarcinoma - Specific Uridine Diphospho - N - Acetylglucosamine α-6-D-Mannoside β-D-1,6-N-Acetylglucosaminyltransferase V

(Cheorl Ho Kim),(Chun Park),(Tae Wook Chung),(Byung Yoon Cha),(Sung Kwon Moon),(Dong Wook Kim),(Young Choon Lee) 한국키틴키토산학회 2002 한국키틴키토산학회지 Vol.7 No.1

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