Conditioned Media of ASC-17D Sertoli Cells Induce G1-Growth Arrest of DU145 Human Prostate Cancer Cells

Min, Bon Hong,Kang, Sang Wook,Lee, Kwang Ho,Choi, Sang Hyun,Kang, Hyeog,Chun, Boe Gwun The Korea Science and Technology Center 1998 BMB Reports Vol.31 No.5

We studied the effects of ASC-17D rat Sertoli cell-conditioned media (rSCCM) on the proliferation of the DU145 prostate cancer cells. rSCCM was prepared from ASC-17D cells cultured in DMEM/F-12 serum-free media at a nonpermissive temperature of 40℃, which is the condition for the high expression of clusterin. We found that rSCCM could inhibit the proliferation of DU145 cells by arresting the cell cycle in the G1 phase in a dose-dependent manner. This growth arresting activity was abolished by boiling rSCCM for 5 min. The G1 growth-inhibiting activity of rSCCM was also detected in other prostate-originated cancer cells examined (ie., LNCaP and PC-3) but not in other cells (ASC-17D, HepG2, SK-N-SH, and NIH3T3). Western blot analysis of partially purified growth inhibiting fractions with the clusterin antibody showed that the cytostatic factor in rSCCM was not clusterin. This cytostatic factor was semipurified by DEAE-Sepharose, ammonium sulfate precipitation, and Phenyl-Sepharose column chromatography, and was estimated to have a molecular weight of 88 kDa by Sephacryl S-300 gel filtration.

Artificial Shrinkage Improves Survival and Embryonic Quality of Vitrified Bovine Blastocysts Derived from IVF and SCNT

Sung-Hun Min,Joo-Hee Hong,Humdai Park,Deog-Bon Koo 한국동물생명공학회(구 한국동물번식학회) 2011 발생공학 국제심포지엄 및 학술대회 Vol.2011 No.1

Freezing of bovine blastocysts has been proposed as a tool to improve the feasibility of cattle production by using embryo transfer technique. However, the low efficiency of frozen-thawed embryos survival and further development is a crucial problem. Thus, we examined the effect of artificial shrinkage before vitrification of bovine expanded, hatched and SCNT embryos on the survival rate, apoptosis index and further development after thawing. Expanded, hatched and SCNT embryos were vitrified after artificial shrinkage, which was performed by puncturing the blastocoele with a pulled pasteur pipet. Artificial shrinkage of the blastocyst was achieved after pushing a pulled pasteur pipet into the blastocoele cavity until it contracted. The shrunken and not shrunken embryos were exposed to cryoprotectant solution in 7.5% ethylene glycol-7.5% DMSOPBS with 20% FBS for 5 min. They were placed in a small volume of vitrification solution (15% ethylene glycol+15% DMSO+PBS+20% FBS+0.5 M sucrose) and plunged into liquid nitrogen on a cryotop. Then, after thawing, cryoprotectant was diluted in 1.0 M, 0.5 M, 0.25 M, and 0 M sucrose for 1, 3, 5, and 5 min. Under the optimal conditions, overall efficiency of the survival rate of bovine expanded, hatched, SCNT embryos in artificial shrinkage groups was higher compared with non-artificial shrinkage groups (p< 0.05). Especially, the numbers of TUNEL-positive nuclei in artificial shrinkage groups were significantly reduced than those of non-artificial shrinkage groups among frozen-thawed expanded, hatched, and SCNT blastocysts (p< 0.05). Our results showed that survival rates in cryopreserved expanded, hatched, SCNT embryos could be improved by reducing the fluid content. Therefore, we suggest that artificial shrinkage method is a effective pretreatment technique for the cryotop vitrification of expanded, hatched, SCNT bovine blastocysts.

Purification and Characterization of a Xylanase from Bacillus sp . KK-1

Dae Joon Kim,Ki Hong Yoon Seung Ho Kim,Ki Haeng Cho,Bon Hong Min 생화학분자생물학회 1995 BMB Reports Vol.28 No.4

A thermophilic Bacillus sp. strain KK-1 isolated from soil produced an extracelluar xylanase. From the culture supernatant of Bacillus sp., the xylanase was purified to homogeneity by ammonium sulfate precipitation and DEAF-Sephadex A-50 chromatography. The molecular weight of the purified xylanase was estimated to be 45 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel permeation chromatography. The apparent K_m values for xylanase, using oat spelt xylan and birchwood xylan as substrates, were 7.1 ㎎/㎖ and 3.2 ㎎/㎖, and V_(max) values were 27.0 μ㏖·min^(-1)·㎎^(-1) and 29.0 μ㏖·min^(-1)·㎎^(-1), respectively. The xylanase hydrolyzed oat spelt xylan to mostly xylobiose, xytotriose, and xylose. The amino acid composition indicated that the xylanase contained high amounts of amino acid residues of glutamic acid and glutamine (Glx) and aspartic acid and asparagine (Asx).

NO(Nitric Oxide)에 의한 이자의 외분비 조절작용

한유미,김병무,민본홍,박인선 대한해부학회 2000 Anatomy & Cell Biology Vol.33 No.5

천추 갈색종양(Brown tumor)으로 발현된 부갑상선 기능항진증 1예

홍우정,구본정,이재민,한상우,이효진,나소영,노흥규,김영건,송민호 충남대학교 의학연구소 2003 충남의대잡지 Vol.30 No.2

부갑상선 기능항진증에 의한 골병변으로 갈색종양이 있다. 갈색종양은 주로 장골을 침범하고 척추골에서의 발생은 매우 드물다. 국내의 보고에서 갈색종양의 장골 침범은 있었으나, 척추골에서의 발생은 보고된바 없다 .저자들은 부갑상선 기능항진증으로 인하여 척추골의 천추에 발생된 갈색종양 1예를 경험하였기에 문헌고찰과 함께 보고하는 바이다. The bony involvement in primary hyperparathyroidism present as browm tumor or osteitis fibrosa cystica and lead to pathologic fracture. Brown tumor commonly involve long bone such as femur, tibia, fibula, humerus, and radius. Brown tumor of axial skeleton is a rare entity. Previously reported case that involve axial skeleton still not present. We report a 47-year-old male patient that he has a hyperparathyroidism presenting as brown tumor involving sacrum.

갑상선 세포에서 전사보조활성인자인 CBP와 CIITA의 인터페론-감마 활성 부위에 대한 조절 : CBP and CIITA on GAS(Interferon- γ Activated Site)-Mediated Transcription in Thyroid Cells

채수흥,홍우정,김군순,조영석,김도희,김현진,구본정,송민호,김영건,노흥규 충남대학교 의과대학 지역사회의학연구소 2001 충남의대잡지 Vol.28 No.1

Background : In the previous stuides, we identified that the interferon-r activated sequence (GAS) in the 5-flanking region of rat ICAM-1 gene is major element for interferon-r-inducible expression of the gene in rat thyroid cells, FRTL-5. We here, investigated the role of transcriptional coactivators, CBP(CREB binding protein) and CⅡTA (class Ⅱ transactivator) in the modulation of the activity of GAS which could interacts with signal transducers and activators of transcription-1 and 3 (STAT1 and STAT3). Methods : The expression of CBP RNA and protein were quantitated in FRTL-5 after stimulation with interferon-r (IFN-r), thyroid stimulating hormone (TSH), forskolin and methimazole. Direct association of CBP with STAT were analyzed by immunoprecipitation. The transcriptional roles of CBP and CⅡTA in the regulation of GAS were assessed by the cotransfection with their expression vectors with reporters; 5-deletion constructs of rat ICAM-1 promoter or 8xGAS-luc constructs, into FRTL-5 thyroid cells. Results : The level of CBP RNA and protein were not changed by the treatment with TSH, IFN-r, forskolin and methimazole in FRTL-5, FRT and BRL liver cells. The CBP could be directly associated with STAT1. Furthermore, the overexpression of CBP significantly increases the both promoter activities; rat ICAM-1 gene promoter which has GAS element and 8xGAS-luc cassette constructs. However, the cotransfection of CⅡTA decreased the constitutive and CBP-mediated transactivation of rat ICAM-1 promoter and 8xGAS-luc cassette constructs. Conclusion : We identified that the two tanscriptional coactivators; CBP and CⅡTA has differential roles in the regulation of transcriptional activity of GAS drived promoter. CBP increases the GAS activity through the direct binding with STAT1, but CⅡTA inhibited the CBP-mediated transactivation of GAS activity.

各種 Alcohol 濃度의 人蔘酒가 Mouse의 腦 및 心臟組織內 Catecholamine 含量에 미치는 影響

申萬鍊,全普權,閔本泓,李在鉉,金鐸 최신의학사 1985 最新醫學 Vol.28 No.10

HMG-CoA 환원효소 억제제에 의한 ICAM-1 유전자의 발현조절

김현진,정효균,홍우정,김군순,조영석,김도희,채수흥,구본정,송민호,노흥규,김영건 충남대학교 의과대학 지역사회의학연구소 2001 충남의대잡지 Vol.28 No.1

Background : ICAM-1 act as one of major adhesion molecules in the atherosclerotic lesion. ICAM-1 expression is mainly regulated at the level of transcription and depend on IFN-γ signal transduction pathway in which the STAT1 transcrption factor is a critical intermediate. IFN-γreceptor not only initiates tyrosine 701 phosphorylation of STAT1 by Jak1 and Jak2, but also phosphorylates serine 727 through the activation of Raf-1/MAP kinases. HMG-CoA reductase inhibitors have anti-atherosclertic effects, beyond normalization of hypercholesterolemia, by directly acting on endothelial cells, macrophages and vascular smooth muscle cells. HMG-CoA reductase inhibitors suppress the synthesis of isoprenoid intermediates such as geranylgeranyl-pyrophosphate or farnesylpyrophosphate. These effects results inhibitors suppress the synthesis of isoprenoid intermediates such as geranylgeranyl-pyrophosphate or farnesylpyrophosphate. These effects results inhibition posttranslational farnesylation and geranyl-geranylation processing of small GTP-binding preoteins and inhibition of normal signaling activities. Method : We made several 5'-deletion constructs of rat ICAM-1 promoter and analyzed the promoter activities by measuring the luciferase activity after transfection into ECV304 cells and smooth muscle cells. We checked the level of total and phosphorylated STAT1 protein by immunoblot analysis using specific antibodies. Results : Lovastatin inhibits IFN-γ-induced ICAM-1 gene expression in the ECV304cell. The cells pretreated with PD98059, MEKK inhibitor showed significantly low ICAM-1 RNA induction with IFN-γ stimulatio. IFN-γ induced phosphorylation of tyrosine 701 was not significantly changed by the pretreatment of lovastatin. But lovastatin suppresses IFN-γ-induced phosphorylation of ERK1/ERK2 which are responsible for the seine 727 phosphorylation in STAT1. Conclusion : We showed that HMG-CoA reductase inhibitors, lovastatin, suppresses IFN-γ mediated ICAM-1 gene expression through the inhibition of transcription. HMG-CoA reductase inhibitor suppresses IFN-γ-induced phosphorylation of serine 727 in STAT1 through the modulation of MAP kinases.

BC₃H1 혈관평활근세포의 증식 및 분화과정에 있어서 Polyamine 대사의 역할에 관한 연구

최상현,문창택,민본홍,전보권,천연숙 고려대학교 의과대학 1992 고려대 의대 잡지 Vol.29 No.3

The influences of spermine (SM) and DFMO. an irreversible ODC-inhibitor, on the polyamine metabolism, CK-activity, [^(3)H]-thymidine DNA synthesis, and isoactin mRNA expression of BC_(3)H1 in culture were studied, with the references of phosphodiesterase(PDE)-inhibitors including isobutyl-methylxanthine (IBMX) and KR30075. BC_(3)Hl cells cultured in 10% fetal bovine serum (FBS)-DMEM for 24 hrs (the first : pre-drug phase) were cultivated in 10% FBS-DMEM added with one of the test drugs for further 96 hrs(the second proliferation phase), and then continue to grow in 2% FBS-DMEM for 48 hrs (the third : differentiation phase). The putrescine content of BC_(3)Hl harvested after the first phase was relatively higher in comparison with those of the other phases and was gradually decreased through the full phases of culture, and the spermine content was little changed. The spermidine (SD) content was. unlike other polyamines, increased by 26.5% at the end of the second phase, but that SD increase was moderately attenuated by PDE-inhibitors, particularly KR30075, and furthermore. the SD content of BC_(3)Hl cultured in the presence of DFMO, unlike other polyamines. showed the marked and gradual decrease throughout the full phases of culture. The [^(3)H]-thymidine DNA synthesis during the third : differentiation phase was not affected by IBMX and slightly inhibited by KR30075, but was significantly inhibited by SM plus DFMO or each of them. And the creatine kinase (CK) activity gradually increased in advancing with culture duration was not changed by PDE-inhibitors. but both SM and DFMO significantly enhanced the increase of CK activity. The synthesis of BC_(3)Hl α-actin and β-/ γ-actin mRNAs was significantly enhaced by DFMO, and the DFMO-induced enhancement was dramatically inhibited by SM. And PDE-inhibitors and SM little affected the synthesis of α-actin mRNA, but PDE-inhibitors attenuated the synthesis of β-/ γ-mRNAs, especially in the second phase. These results suggest that polyamines have a pivotal role in the proliferation and differentiation of BC_(3)Hl cells, that the enhancement induced by DFMO of expression of VSM isoactin gene seems to be an attractive subject to be studied in future, and that PDE-inhibitors may inhibit the synthesis of VSM β-/ γ-actin mRNAs.

부신 결핵에 의한 Addison씨병 1예

송치운,구본정,안봉수,전준식,안미애,이진홍,송민호,김영건,노흥규 충남대학교 의과대학 지역사회의학연구소 1995 충남의대잡지 Vol.22 No.2

Addison's disease is a rare primary adrenal insufficient disorder resulting from chronic deficiency of adrenal cortical hormones. Clinical manefestations are generalized weakenss, weight loss, hyperpigmentation (especially sun exposed area and mucous membrane), hypotension, hyponatremia, hyperkalemia, gastrointestinal symptoms (involving anorexia, nausea, vomiting, abdominal pain). A 34-year-old woman has experienced slowly progressive generalized weakenss and skin pigmentation, anorexia, nausea, vomiting with ascites and diffuse abdominal pain. On the time of admission, her main clinical manifestations were anorexia, nausea, vomiting, fatigue, generalized weakness, amenorrhea, hair loss, diffuse abdominal pain revealed as Addison's disease due to bilateral adrenal tuberculosis. Her adrenal insufficient symptoms were recovered with the replacement of adrenocortical hormones and antituberculous medications. After treatment, Her skin pigmentation was decreased and menstruation was reappeared. Here we experienced one cases of Addison's disease with tuberculous peritonitis.