일차배양 Sertoli cell 에서 Testosterone 에 의한 세포성 핵암유전자 발현에 관한 연구

임규,유지헌,김계영,권기량,황병두 ( Kyu Lim,Ji Hun Yoo,Kye Young Kim,Gi Ryang Kweon,Byung Doo Hwang ) 생화학분자생물학회 1993 BMB Reports Vol.26 No.1

The expression of the nuclear protooncogenes have been associated with the cell proliferation and differentiation into changes in nuclear function. To evaluate the possibility that the nuclear protooncogenes play roles testosterone-dependent gene regulation, the effects of testosterone on the expression of the nuclear protooncogenes have been investigated in primary Sertoli cell cultures. c-Myc, c-jun and c-fos were expressed in steady state of primary Sertoli cells but c-myb was not detected. Testosterone increased c-myc and c-jun mRNA with maximal stimulation reached in 16 h. The induction of c-myc was dependent on the concentrations of testosterone. The testsoterone induced c-myc mRNA level was also increased in the cells treated with cycloheximide, but reduced by actinomycin D pretreatment. Even in the absence of testosterone, c-myc mRNA was clearly detectable in Sertoli cells from 8-days-old rats, but hardly detectable in cells from 14 and 28 days of age. Testosterone stimulated c-myc expression in the Sertoli cells from 8 and 14-days-old rats. These results suggest that transient expression of c-myc may play some roles to an intermediate step required for testosterone-dependent regulation of other genes expression of Sertoli cells.

일차배양 Sertoli Cell에서 Testosterone에 의한 세포성 핵암유전자 발현에 관한 연구

임규,유지헌,김계영,권기량,황병두,Lim, Kyu,Yoo, Ji-Hun,Kim, Kye-Young,Kweon, Gi-Ryang,Hwang, Byung-Doo 생화학분자생물학회 1993 한국생화학회지 Vol.26 No.1

일차배양 Sertoli cell에서 testosterone에 의한 c-myc 등 세포성 핵암유전자 발현 및 그 조절기전을 검색하여 다음과 같은 결과를 얻었다. Steady state에서 발현되는 Sertoli cell 세포성 핵암유전자는 c-fos, c-myc, c-jun이었으며 c-myb은 검출되지 않았고 이때 mRNA 크기는 각각 2.2 kb, 2.4 kb, 2.7 kb에 해당되었다. Testosterone에 의한 c-myc 및 c-jun mRNA의 발현은 시간경과에 따라 증가하였으며 유도되는 시간은 모두 16시간에 최고치에 달하여 유사하였다. c-Myc은 testosterone 농도에 비례해서 그리고 cycloheximide 처리로 각각 전사가 증가되었으나 actinomycin D 처리로 억제되었다. 연령에 따른 Sertoli cell에서의 c-myc mRNA 발현에 대한 testosterone의 영향은 steady state에서는 8일령이 가장 높았으며, testosterone에 의한 유도는 8일령, 14일령에서는 나타났으나 28일령에서는 유도되지 않았다. 이상의 결과로 미루어 보아 testosterone에 의해 유도된 c-myc mRNA가 testosterone에 의해 조절되는 Sertoli cell의 다른 유전자 발현의 조절에 관련되어 있을 것이라 시사된다. The expression of the nuclear protooncogenes have been associated with the cell proliferation and differentiation into changes in nuclear function. To evaluate the possibility that the nuclear protooncogenes play roles testosterone-dependent gene regulation, the effects of testosterone on the expression of the nuclear protooncogenes have been investigated in primary Sertoli cell cultures. c-Myc, c-jun and c-fos were expressed in steady state of primary Sertoli cells but c-myb was not detected. Testosterone increased c-myc and c-jun mRNA with maximal stimulation reached in 16 h. The induction of c-myc was dependent on the concentrations of testosterone. The testsoterone induced c-myc mRNA level was also increased in the cells treated with cycloheximide, but reduced by actinomycin D pretreatment. Even in the absence of testosterone, c-myc mRNA was clearly detectable in Sertoli cells from 8-days-old rats, but hardly detectable in cells from 14 and 28 days of age. Testosterone stimulated c-myc expression in the Sertoli cells from 8 and 14-days-old rats. These results suggest that transient expression of c-myc may play some roles to an intermediate step required for testosterone-dependent regulation of other genes expression of Sertoli cells.

졸겔법으로 합성된 Mal₂O₄:Eu²⁺(M=Sr,Ba,Ca)의 특성

임규,김용현,이형직,김세기,이형복,Lim, Kyu,Kim, Young-Hyun,Lee, Hyung-Jik,Kim, Sei-Ki,Lee, Hyung-Bock 한국세라믹학회 2009 한국세라믹학회지 Vol.46 No.6

Phosphors of $Eu^{2+}$ doped alkaline earth aluminates MA$l_2O_4:Eu^{2+}$ (M=Sr, Ba, Ca) have been prepared by sol-gel process and their characterization of photoluminescence and photocurrent properties have been performed. The phosphors prepared by sol-gel process, due to its advantage of better homogeneity and low synthetic temperature, was synthesized as single phases at lower temperature than the solid-state process; $800{\sim}1000{^{\circ}C}$ for 6 h under mild reduction atmosphere (Ar- 3% $H_2$). It was confirmed that SrA$l_2O_4:Eu^{2+}$ composition revealed the most excellent properties in the brightness and photocurrent.

사람 태반조직 DNA Topoisomerase Ⅰ에 관한 연구

이정복,권기량,임규,황병두 ( Jeong Bok Lee,Gi Ryang Kweon,Kyu Lim,Byung Doo Hwang ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.4

DNA topoisomerase I has been purified from human term placenta approximately 520-folds with a 10% yield. The enzyme shows a broad pH optimum from pH 6 to 9 and is heatlabile, being completely inactivated by heat treatment at 50℃ for 5 min. The enzyme is activated by K^+ and Na^+ approximately 20 and 8-folds respectively. Magnesium ion (Mg^(2+)) is the most potent activator, the activity being 20 and 40-folds activated at 2 mM and 10 mM respectively. But copper ion (Cu^(2+)) is a potent inhibitor. In the presence of Mg^(2+) and K^+, the enzyme is inhibited by physiologic concentration of ATP and GTP. Inhibitory mechanism of ATP is considered to be a inhibition of readoptation of active enzyme conformation and that of GTP is to be a inhibition of the prior step of DNA rejoining. The molecular weight is around 68,000. Camptothecin and 10-hydroxycamptothecin inhibit this enzyme, and the inhibitory action of 10-hydroxycamptothecin is potentiated by Mg^(2+) and K^+. DNA fragmentation by 10-hydroxycamptothecin is more potent than that by camptothecin in the DNA cleavage assay. Heparin and Cu^(2+) inhibit the prior step of DNA rejoining by this enzyme. From the above results, the inhibitory action mechanism of ATP, GTP, heparin, Cu^(2+) and the possible role of Mg^(2+) and K^+ for potentiating the inhibitory action of 10-hydroxycamptothecin, and the development the anticancer drug against DNA topoisomerase I as a target enzyme are discussed.

가감보양환오탕(加減補陽還五湯)의 N2a 뇌신경세포에 대한 보호효과

박용기 ( Yong Ki Park ),임규 ( Kyu Lim ) 대한본초학회 2006 大韓本草學會誌 Vol.21 No.4

인체 혀의 편평세포암 세포에서 ω3-fatty acid desaturase 유전자 발현이 침윤 및 종양형성에 미치는 영향

홍태화(Tae-Hwa Hong),신소연(Soyeon Shin),한승현(Seung-Hyeon Han),황병두(Byung-Doo Hwang),임규(Kyu Lim) 한국생명과학회 2018 생명과학회지 Vol.28 No.8

오메가-3 지방산(오메가-3)은 수종의 암에 대해 종양형성 억제 및 침윤이 억제됨이 알려져 있다. 그러나 혀의 편평세포암 세포에서 내인성 오메가-3에 의한 침윤 및 종양형성 억제 대한 연구가 명확하게 보고된 바 없다. 이에 본 연구는 혀의 편평세포암 세포에서 ω3-fatty acid desaturase의 유전자 발현이 침윤 및 종양형성에 미치는 영향을 규명하였다. 먼저 SCC-4 및 SCC-9세포의 침윤능은 오메가-3인 DHA 처리에 의해 억제 됨을 확인 하였다. DHA 처리 후 MMP-9 및 MMP-2 활성이 감소 되었을 뿐만 아니라 그 promoter의 reporter 활성도 억제하였다. 또한 COX-2 및 VEGF promoter 활성 뿐만 아니라 NF-kB 활성도 DHA에 의해 억제 되었다. SCC-9의 ω3-desaturase 유전자 stable 세포(fSCC-9sc)의 세포증식 및 colony formation이 억제 되었으며, in vivo 동물실험에서 fSCC-9sc 세포의 종양형성능은 현저히 억제 되었고, 면역형광염색법을 이용한 fSCC-9sc 세포의 종양 조직에서의 TUNEL 양성세포는 대조군인 fSCC-9cc 세포에 비해 현저히 증가하였다. 이상의 결과로 오메가-3는 인체 혀의 편평세포암 세포의 침윤 뿐만 아니라 종양형성을 억제하여 항암작용을 나타낼 수 있으며 따라서 오메가-3는 인체 혀의 편평암의 예방 및 치료에 유용하게 사용될 수 있으리라 생각된다. Omega-3 polyunsaturated fatty acids (ω3-fatty acid) have been found to possess anticancer properties in a variety of cancer cell lines and animal models, but their effects in human tongue squamous cell carcinomas (SCCs) remain unclear. This study was designed to examine the effect of ω3-fatty acid desaturase (fat-1) gene expression on invasion and tumorigenicity in human tongue SCC cells and the molecular mechanism of its action. Docosahexaenoic acid (DHA) treatment inhibited in vitro invasion in a dose-dependent manner. In zymography, matrix metalloproteinase-9 (MMP-9) and Matrix metallopeptidase-2 (MMP-2) activities were reduced, and MMP-9 and MMP-2 promoter activities were inhibited by the DHA treatment. In addition, cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) promoter reporter activities were inhibited in SCC-4 and SCC-9 cells after the DHA treatment. To investigate the effect of a high level of endogenous ω3 fatty acids, a stable SCC-9 cell line expressing the ω3-desaturase gene (fSCC-9sc) was generated. The growth rate and colony-forming capacity of fSCC-9sc were remarkably decreased as compared with those of fSCC-9cc. Likewise, the tumor size and volume of fSCC-9sc implanted into nude mice were significantly inhibited, with increases in the cell death index. Furthermore, a transwell chamber invasion assay showed a reduction in cell invasion of the fSCC-9sc lines when compared with that of the fSCC-9cc line. These findings suggested that fat-1 gene expression inhibited tumorigenicity, as well as invasion in human tongue SCC cells. Thus, utilization of ω3 fatty acids may represent a promising therapeutic approach for chemoprevention and the treatment of human tongue SCCs.

TALP-32의 인체자궁암 세포주 HeLa에 대한 세포독성

박지훈(Ji-Hoon Park),김종석(Jong-Seok Kim),윤은진(Eun-Jin Yun),송경섭(Kyoung-Sub Song),서강식(Kang-Sik Seo),김훈(Hoon Kim),정연주(Yeon-Joo Jung),윤완희(Wan-Hee Yun),임규(Kyu Lim),황병두(Byoung-Doo Hwang),박종일(Jong-Il Park) 한국독성학회 2006 Toxicological Research Vol.22 No.4

TALP-32 is highly basic protein with a molecular weight of 32 kDa purified from human term placenta. Some basic proteins such as defensins and cecropins are known to induce cell death by increasing membrane permeability and some of them are under development as an anticancer drug especially targeting multi-drug resistant cancers. Therefore, we investigated cytotoxic effect and mechanism of TALP-32. When HeLa cell was incubated with TALP-32, cytotoxicity was increased in time and dose dependent manner. As time goes by, HeLa cells became round and plasma membrane was ruptured. Increase of plasma membrane permeability was determined with LDH release assay. Also in transmission electron microscopy, typical morphology of necrotic cell death, such as cell swelling and intracellular organelle disruption was observed, but DNA fragmentation and caspase activation was not. And necrotic cell death was determined with Annexin V/PI staining. The cytotoxicity of TALP-32 was minimal and decreased on RBC and Hep3B respectively. These data suggests that TALP-32 induces necrosis on rapidly growing cells but not on slowly growing cells implicating the possibility of its development of anticancer peptide drug.

히스티딘의 IgE 매개 알레르기 반응 억제 효과

실왈 프라산타(Prashanta Silwal),최슬기(Seulgi Choi),신근아(Keuna Shin),이찬용(Chan Yong Lee),박종일(Jong Il Park),허준영(Jun-Young Heo),임규(Kyu Lim),박승길(Seung-Kiel Park) 한국식품영양과학회 2018 한국식품영양과학회지 Vol.47 No.7

비만세포는 알레르기를 유발하는 중요한 세포로서 항원의 자극을 받으면 알레르기 유발 물질을 분비한다. 히스티딘은 비만세포가 관여하는 만성 장염 증세를 완화하는 기능이 있다. 그러나 히스티딘이 비만세포의 활성화에 미치는 영향에 대한 연구는 없다. 우리는 알레르기 반응 연구의 동물모델인 IgE 매개 수동 피부 아나필락시스 방법 그리고 비만세포가 알레르기 유발 물질을 분비하는 반응(탈과립 반응, 염증성 지질 및 사이토카인 분비)을 통해 히스티딘이 알레르기에 미치는 영향을 연구하였다. 생쥐의 복강으로 히스티딘을 100 mg/kg으로 투여하면 수동 피부 아나필락시스를 통계적으로 의미 있는 수준으로 억제하였고, 비만세포에서도 탈과립 반응과 알레르기 유발 불질의 분비를 억제하였다. 이러한 결과는 히스티딘 섭취는 IgE 매개 알레르기를 억제하는 데 유익할 것으로 생각할 수 있다. Mast cells are major immune cells in allergies that secrete allergic mediators in response to antigen stimulation. The amino acid, histidine, ameliorates the inflammation that mast cells are involved. On the other hand, there are no reports of the effects of histidine on the activation of mast cells. This study examined the anti-allergic effects of histidine with well-established experimental methods in allergy studies, including IgE-mediated passive cutaneous anaphylaxis (PCA) in mice and mast cell activation responses, such as degranulation, secretion of inflammatory lipid LTB4 and cytokines (TNF-α and IL-4). The intraperitoneal administration of 100 mg/kg histidine suppressed PCA significantly (P<0.05) in mice. In addition, histidine inhibited IgE-mediated mast cell activation, including degranulation and the production of LTB4 and the inflammatory cytokines, TNF-α and IL-4. Overall, histidine suppressed the IgE-mediated allergic responses in vivo and in vitro. Histidine supplementation is expected to beneficial for IgE-mediated allergies.

Tryptophan Negatively Regulates IgE-mediated Mast Cell Activation

Prashanta Silwal(실왈 프라산타),Keuna Shin(신근아),Seulgi Choi(최슬기),Uk Namgung(남궁욱),Chan Yong Lee(이찬용),Jun-Young Heo(허준영),Kyu Lim(임규),Jong IL Park(박종일),Ki-Hwan Kim(김기환),Seung-Kiel Park(박승길) 대한체질인류학회 2017 해부·생물인류학 (Anat Biol Anthropol) Vol.30 No.2

비만세포는 알레르기 반응을 일으키는 주된 세포로서 항원 자극에 반응하여 알레르기 유발 물질인 히스타민, 단백질 분해효소, 염증성 지질 물질 및 사이토카인 등을 분비한다. 아미노산인 트립토판은 염증반응을 조절한다. 트립토판 투여는 비만세포가 관여하는 염증성 장염 증상을 완화시킨다. 그러나 트립토판이 비만세포의 알레르기 반응에 미치는 영향에 대한 연구는 없다. 본 저자들은 트립토판이 IgE 매개 알레르기 반응에 미치는 영향을 비만세포 수준에서 그리고 실험동물 생쥐에서 연구하였다. IgE-매개 수동 피부 아나필락시스를 생쥐에서 연구하였다. 또한 IgE-매개 비만세포 활성화 반응 즉, 탈과립 반응, 염증성 지질인 LTB4와 사토카인 (TNF-α와 IL-4) 등의 분비를 측정하였다. 트립토판을 생쥐에 복강 주사하면 IgE 매개 수동 피부 아나필락시스를 억제하였다. 또한 비만세포 수준에서도 트립토판은 IgE 매개 알레르기 반응들, 즉 탈과립 반응과 염증성 지질인 LTB4 및 사이토카인인 TNF-α와 IL4의 분비를 억제하였다. 이러한 결과로부터 트립토판은 IgE 매개 알레르기 반응을 세포 수준 및 실험동물 수준에서 억제함을 알 수 있었다. Mast cells are major immune cells in allergy to secrete allergic mediators by a degranulation process and make and secrete inflammatory lipids and cytokines in response to antigen stimulation. An amino acid tryptophan regulates immune functions. Tryptophan ameliorates inflammatory colitis in which mast cells are engaged. However, its effects on mast cells remain to be solved. We investigated the effect of tryptophan on IgE-mediated allergic responses in the mast cells and mice. IgEmediated passive cutaneous anaphylaxis (PCA) in mice were examined. Also IgE-mediated mast cell activation responses such as degranulation of stored granules and secretion of inflammatory lipid LTB4 and cytokines (TNF-α and IL-4) were measured. Intraperitoneal administration of tryptophan suppressed PCA in mice. Also, in the cellular level tryptophan inhibited IgE-mediated mast cell activation such as IgE-mediated degranulation and the production of LTB4. Also, it inhibited production of inflammatory cytokines TNF-α and IL-4. In summary, tryptophan suppressed IgE-mediated allergic activation in vivo and in vitro. Tryptophan supplementation is beneficial for IgE-mediated allergy.

백서의 일차배양 간세포에서 Plasminogen Activator와 Plasminogen Activator Inhibitor의 표피 성장인자 의존적 발현에 관한 연구

윤완희(Wan-Hee Yoon),최정훈(Jeong-Hun Choi),정연주(Yeon-Joo Jung),이병학(Byung-Hak Lee),김태동(Tae-Dong Kim),송인상(In-Sang Song),배진선(Jin-Sun Bae),임규(Kyu Lim),황병두(Byung-Doo Hwang) 대한외과학회 2001 Annals of Surgical Treatment and Research Vol.61 No.3