The Relationship between Intracellular Protein Kinase C Concentration and Invasiveness in U-87 Malignant Glioma Cells

지철,조경근,이경진,박성찬,조정기,강준기,최창락,Ji, Cheol,Cho, Kyung-Keun,Lee, Kyung Jin,Park, Sung Chan,Cho, Jung Ki,Kang, Joon Ki,Choi, Chang Rak The Korean Neurosurgical Society 2001 Journal of Korean neurosurgical society Vol.30 No.3

교모세포종은 비교적 흔한 원발성 뇌종양이며 생물학적 특성상 빠른 성장률을 보이는 것 외에 침습성이 강하여 종양과 인접한 부분을 파괴 시킬 뿐 아니라 직접접촉하지 않는 부분의 파괴도 일어나게 되어 그 결과 치료 예후가 매우 불량한 것으로 되어 있다. 이러한 불량한 예후를 개선 시키기 위해서는 이들 종양의 침습에 대한 기전의 정확한 이해가 필요하며 이를 이용한 새로운 치료방법이 요구된다할 것이다. Protein kinase C(PKC)는 세포내 신호전달체제 과정에서 매우 중요한 역할을 하는 효소로 세포막 수용체 신호를 핵으로 전달하는 역할을 하며 세포내 여러 생물학적 작용이 알려져 있다. 본 실험은 종양침습과 연관하여 세포내 PKC가 어떠한 작용을 하는지에 대해서 악성교종 세포를 대상으로 하여 알아보고자 하였다. 따라서 PKC가 종양침습에 중요한 역할을 할 것이라는 가설을 세웠고 이 가설을 증명하기 위해 세포내 PKC농도를 길항제 및 촉진제를 이용하며 높고 낮게 조절함으로써 그에 따른 침습성의 변화를 살펴보았다. 방법으로는 교모세포종 세포주인 U-87 세포를 약제로 처리한 후 인위적으로 조절된 세포내의 PKC에 대해 효소의 활성도를 측정하였고 침습성은 matrigel artificial basement membrane assay 및 tumor spheroid fetal rat brain aggregate(FRBA) confrontation assay를 이용하여 측정하였다. 결과로 PKC의 길항제인 tamoxifen과 hypericin으로 처치한 세포는 PKC의 활성과 침습도가 모두 감소하였으며 이는 약제농도에 비례하여 나타났다. 반면 PKC 자극제인 TPA로 처치된 세포는 증가된 PKC 활성도나 침습도을 보이지 않았다. 이러한 결과를 종합해 보았을 때 PKC는 종양세포의 침습성에 중요한 역할을 함을 알 수 있었으며 PKC의 길항제는 종양 치료에 유용한 화학 요법 제가 될 수 있을 것으로 사료된다. Objective : Glioblastomas, the most common type of primary brain tumors, are highly invasive and cause massive tissue destruction at both the tumor invading edges and in areas that are not in direct contact with glioma cells. As a result, patients with high-grade gliomas are faced with a poor prognosis. Such grim statistics emphasize the need to better understand the mechanisms that underlie glioma invasion, as these may lead to the identification of novel targets in the therapy of high grade gliomas. Protein kinase C(PKC) is a family of serine/threonine kinases and an important signal transduction enzyme that conveys signals generated by ligand-receptor interaction at the cell surface to the nucleus. PKC appears to be critical in regulating many aspects of glioma biology. The purpose of this study was to assess accurately the role of PKC in the invasion regulation of human gliomas based on hypothesis that protein kinase C(PKC) is functional in the process of glial tumor cell invasion. Method : To test this hypothesis, U-87 malignant glioma cell line intracellular PKC levels were up and down regulated and their invasiveness was tested. Intracellular PKC level was characterized using PKC activity assays. Invasion assays including barrier migration and spheroid confrontation were used to study the relationship between PKC concentration and invasiveness. Result : The cell line which were treated by PKC inhibitor tamoxifen and hypericin exhibited decreased PKC activity and decreased invasive abilities dose dependently both in matrigel invasion assay and tumor spheroid fetal rat brain aggregates(FRBA) confrontation assay. However, the cell line that was treated by PKC activator 12-O-tetradecanylphorbol-13acetate(TPA) did not exhibit increases in either PKC activity or invasive ability. Conclusion : These studies suggest that PKC may be a useful molecular target for the chemotherapy of glioblastoma and other malignancies and that a therapeutic approach based on the ability of PKC inhibitors may be helpful in preventing invasion.

실험관 및 생체내에서 U-87 교모세포종 세포주의 광역학 치료 효과에 대한 연구

조우진,조경근,지철,박성찬,박해관,강준기,최창락,Cho, Woo Jin,Cho, Kyung-Keun,Ji, Cheol,Park, Sung Chan,Park, Hea Kwan,Kang, Joon Ki,Choi, Chang Rak 대한신경외과학회 2001 Journal of Korean neurosurgical society Vol.30 No.5

Objective : The objective of this study was to determine the photodynamic therapeutic response of U-87 human glioma cell in vitro as well as in the nude rat xenograft model using photofrin as photosensitizer. Material and Method : U-87 cells were cultured on 96-well culture plates, photofrin(Quadralogic Technologies Inc., Vancouver, Canada) was added into the cell culture medium at concentration of $1{\mu}g/ml$, $2.5{\mu}g/ml$, $5{\mu}g/ml$, $10{\mu}g/ml$ and $20{\mu}g/ml$. 24 hour after drug treatment, cells were treated with optical(632nm) irradiation of $100mJ/cm^2$, $200mJ/cm^2$ and $400mJ/cm^2$. Photofrin(12.5mg/kg, i.p.) was administered to 28 nude rats containing intracerebral U-87 human glioma as well as 26 normal nude rats. 48 hours after administration, animals were treated with optical irradiation(632nm) of $35J/cm^2$, $140J/cm^2$ and $280J/cm^2$ to exposed tumor and normal brain. The photofrin concentration was measured in tumor and normal brain in a separate population of animals. Results : By MTT assay, there was 100% cytotoxicity at any dose of photofrin with optical irradiation of $200mJ/cm^2$ and $400mJ/cm^2$. But at the optical irradiation of $100mJ/cm^2$ cells were killed in dose dependent manner 28.5%, 49.1%, 54.4%, 78.2%, and 84.6% at concentration of $1{\mu}g/ml$, $2.5{\mu}g/ml$, $5{\mu}g/ml$, $10{\mu}g/ml$ and $20{\mu}g/ml$, respectively. Dose dependent PDT lesions in both tumor and normal brain were observed. In the tumor lesion, only superficial tissue damage was found with optical irradiation of $35J/cm^2$. However, in the optical irradiation group of $140J/cm^2$ and $280J/cm^2$ the volume of lesions was measured of $7.2mm^3$ and $14.0mm^3$ for treatment at $140J/cm^2$ and $280J/cm^2$, respectively. The U-87 bearing rats showed a photofrin concentration in tumor tissue of $6.53{\pm}2.16{\mu}g/g$, 23 times higher than that found in the contralateral hemisphere of $0.28{\pm}0.15{\mu}g/g$. Conclusion : Our data indicate that the U-87 human glioma in vitro and in the xenografted rats is responsive to PDT. At these doses, a reproducible injury can be delivered to human glioma in this model. Strategies to spare the normal brain collateral damage are being studied.

전이성 뇌흑색종: 증례보고 및 한국문헌고찰

최승진 ( Seung Jin Choi ),지철 ( Cheol Ji ),안재근 ( Jae Geun Ahn ),민기옥 ( Ki Ouk Min ),김문찬 ( Moon Chan Kim ) 대한뇌종양학회 2004 대한뇌종양학회지 Vol.3 No.1

Malignant melanoma is a rare cancer in Korea as opposed to in Western countries, but its incidence is beginning to rise. The high tendency of a brain metastasis is a characteristic feature of this cancer and that is considerably related to the patitnt`s prognosis. However, a brain metastasis as a result of result of this tumor has been rarely reported in Korea particularly in the aspect of the specific treatment modalities and their outcomes. With reporting a patient with multiple brain metastases of malignant melanoma accompanied with a tumoral hemorrhage, the authors suggest that more aggressive trials of various treatments as well as an estimation of the outcomes will be needed despite the fact that a current multidisciplinary treatment has not yet increased the long term survival.

포스터 전시 : 위장관 ; 장결핵과 구별하기 어려운 Crohn병 1례

주임관 ( Im Kwan Jhu ),양승호 ( Seung Ho Yang ),지황룡 ( Hwang Yong Ji ),손태용 ( Tae Yong Son ),구철 ( Cheol Koo ),윤지영 ( Ji Young Yoon ),정종길 ( Jong Gil Jeong ) 대한소화기학회 2002 대한소화기학회 춘계학술대회 Vol.2002 No.-

<증례> 52세 남자환자가 1주일동안 지속된 우하복부 동통을 주소로 내원하였다. 환자는 8개월 전에도 동일한 증상으로 본원에 내원하여 복강경하 담낭절제술, 충수절제술, 회장말단부 조직검사를 시행하였다. 조직검사에서 만성담낭염이 있었고, 충수돌기는 정상이었다. 회장말단부는 형태가 불분명한 육아종, 염증반응, 그리고 광범위한 섬유화가 보였다. 환자는 추적 관찰하던 중 상태가 좋아져 자의로 추적관찰을 중단하였으며, 증상이 재발하여 내원하였다. 내원당시 신체검

Dieulafoy 궤양과 유사한 복부 방선균증

주임관 ( Ju Im Gwan ),양승호 ( Yang Seung Ho ),최준영 ( Choe Jun Yeong ),구철 ( Gu Cheol ),지황룡 ( Ji Hwang Lyong ),최민석 ( Choe Min Seog ) 대한내과학회 2003 대한내과학회지 증례 특집호 65-5 부록3 Vol.0 No.-

Actinomycosis is a chronic suppurative and granulomatous disease caused by Actinomyces species and characterized by sulfur granule formation. Depending on the site of primary infection, it is generally classified as cervicofacial, thoracic and abdominal t

포스터 전시 : 간 ; 만성 염증성 탈수초성 신경병증 환자에서 스테로이드 치료후 간경병증 발생

양승호 ( Seung Ho Yang ),주임관 ( Im Kwan Jhu ),구철 ( Cheol Koo ),손태용 ( Tae Yong Son ),지황룡 ( Hwang Yong Ji ),신기춘 ( Gi Chun Sin ) 대한소화기학회 2002 대한소화기학회 춘계학술대회 Vol.2002 No.-

<증례> 24세 남자가 2주 전부터 시작된 사지마비와 이상감각을 주소로 내원하였다. 과거력상 B형 간염 보균자였으나 특이한 증세는 없었다. 내원시 혈압 120/80 mmHg, 맥박 76회/분, 호흡수 20회/분, 체온 36.7℃ 이었다. 의식은 명료하였으나 결막 창백이나 공막 황달은 없었다. 호흡음은 정상이었고 호흡곤란이나 심잡음은 없었다. 복부에서 간장이나 비장은 촉지되지 않았다. 입원 당시 시행한 일반혈액검사에서 백혈구 3,500/㎣, 혈색소 13.

충주 수력 부분방전 진단 시스템 신뢰성 고찰

옥연호(Ok Yeon-Ho),임재일(Lim Jae-Il),박지군(Park Ji-Kun),곽원구(Kwak Won-Ku),이재흥(Lee Jae-Heung),최형철(Choi Hyeong-Cheol),이남형(Lee Nam-Hyung),신병철(Shin Byoung-Chol) 대한전기학회 2009 대한전기학회 학술대회 논문집 Vol.2009 No.7

진단기술은 기기의 열화 현상 및 이상 현상으로 진전되는 특성을 측정하여 향후 발생이 예상되는 이상을 조기에 발견하는 것을 목적으로 한다. 기기를 정밀 진단, 분석하여 잔여 수명을 평가하고 적절한 대체계획을 수립함으로써 신뢰도 향상 및 경제적인 설비 운용이 가능하다. 본 논문에서는 수력 발전기 운전 중 부분방전 진단 시스템을 이용하여 수력 발전기의 운전 중에 발생하는 부분방전 신호를 실시간으로 측정, 분석하고 수력 발전기 권선의 절연 상태를 감시 진단하여 기기의 열화 및 이상 현상등을 조기에 발견하고 발전설비의 신뢰도를 향상시킬 수 있는 방안을 제시하고자 한다.

성상세포종에서 표피성장인자수용체의 발현

박영섭,지철,나형균,최창락 대한신경외과학회 1996 Journal of Korean neurosurgical society Vol.25 No.1

It has been demonstrated that the epidermal growth factor receptor(EGFR) gene, the normal human counterpart of the viral orb-B oncogene, is amplified and overexpressed in 40~50% of malignant astrocytomas. Although little is known about the functional status of the overexpressed protein molecule, overexpression of a growth factor receptor evenly distributed throughout a tumor would be an ideal target for monoclonal antibody or growth factor receptor targeted therapy. We undertook the immunohistochemical study of the EGFR in 63 cases of astrocytoma, including 48 cases of malignant astrocytoma, and then compared staining pattern of EGFR with malignancy grade. The results were as follows 1) Expression of EGFR was deteced in 1 cass(6.7%) of low grade astrocytoma, 14 cases(63.6%) of anaplastic astrocytoma and 19 cases(73.1%) of glioblastoma multiforme. It was more frequent in malignat astrocytoma than low grade astrocytoma(P<0.01). 2) The distributed proportion and stains density of EGFR-expressed tumor cells was more increased in glioblastoma multiforme than anaplastic astrocytoma. 3) Regional heterogeneity of EGFR-expressed tumor cells was recognized in cases of EGFR expressed malignant astrocytoma. These results suggest that overexpression of EGFR would be involved in malignant progression of astrocytoma, and the use of monoclonal antibody or growth factor receptor targeted therapy maybe limited due to heterogeneity of EGFR expressed tumor cells in malignant astrocytoma.

흉추 손상 환자의 임상분석

나형균,지철,최창락 대한신경외과학회 1994 Journal of Korean neurosurgical society Vol.23 No.11

We analyzed 10 cases who had received surgery for acute injuries of the thoracic spine complicated by paralysis. Of those 10 cases, 4 patients had a complete and remaining 6 had an incomplete lesion of the spinal cord. Of the 4 patients who had a complete lesion of the spinal cord, significant neurologic function did not recover in 3, regardless of the type and timing of the surgery. In one of 4 patients who had a complete lesion, there was some improvement of sensation. Of the 6 patients who had an incomplete lesion of the spinal cord, 4 received a surgery within 24 hours of injury, and remaining 2 had surgery on posttrauma 30th and 35th days respectively. Average neurologic improvement was 1.75 Frankel grades per patient in the early surgery group and was 1 Frankel grade in the delayed surgery group. Of 6 patients with incomplete lesion, 4 were treated with laminectomy and fusion with instrumentation, and they improved by average decompression and fusion respectively and they improved by 1 Frankel grade respectively. Based on the results of this study and other references, we suggest that early surgical intervention improves neurologic recovery in comparison to late surgical intervention and that posterior surgical instrumentation is indicated in acute unstable bursting fracture and flexion-dislocation injury. Also we suggest that anterior transthoracic decompression and fusion is indicated in cases of neural compression by bone or disc fragments.

벌꿀제품에 있어서 잔류항생물질에 관한 조사연구

구용의,강민철,이선화,김순한,김재이,김은정,김연희,유병옥,김주원,최계선,이향미,지영애,홍무기 식품의약품안전청 2000 식품의약품안전청 연보 Vol.4 No.-

꿀벌의 무저병을 치료하기 위하여 테트라싸이클린 계열의 항생물질(TCs)이 널리 사용되고 있으며,이러한 사용은 벌꿀중 Cs의 잔류문제를 일으킬 수 있다. 따라서 벌꿀중 잔류하는 TCs의 분석법이 팔요하며, 본 연구에서는 벌꿀클의 옥시테트라싸이클린(OTC), 테트라싸이클린fC), 클로르테트라싸이 클린(CTC), 독시싸이클린(DC)끌 분석하기 위해 두가지 HPLC방법을 사용하였다. 아카시아꿀, 잡화꿀,토종꿀, 방꿀등 4가지 종류의 댈꿀에 대한 회수율 및 검출한계를 측정하였다. 형광검출기를 이웅한 분석법에서는 0.Sppm의 수준에온 회수율은 OTC는 91.4~98.2%, TC는 84.B~94.4%, CTC는 뚬.I~105.9%, DC는 뽀.5~져.5%이었으며, 검출한계는 OTC와 TC는 0.01ppf CTC차 OC는 0.05ppm이었다. 자외선검출기를 이용한 분석법에퍼는 0.Sfpm의 수준에서 회수율은 OTC는 78.3~97.0%, TC는 76.8~86.8%, CTC는 89.6~99.5%, Df는 91.I~93.8%이었으며, 검출한계는 OTC와 TC는 0,01ppm, CTC와 DC는 0.02ppm이었다. 두 ITPLC법으로 국내산 벌꿀 140건과 외국산 벌꿀 29건을 조사한 결과 OTC,TC, CTC, BC 모두 검출되지 않았다. ln order to prevent foul brood of honey-bees, tetracycline antibiotics(TCs) are widely used in honey-bee culture. Such usage may resu)t in residues of TCs in honey.Therefore, the establishment of a method for residue analysis of TCs in honey is required.Two HPLC methods for oxytetracycline(OTC), tetracycline(TC), chlortetracycline(CTC) anddoxycycline(DC) in honey have been established. The recoverles of OTC, TC, CTC and BCfrom 4 kinds of honelr(acacia honey, mixed flower honey, native honey, chestnut honey) spikedat a level of 0.Sppm by HPLC with fluorescence detection are 91.4 ~98.2%, 84.B~94.4%, 85.1 ~105.9%, 86.5 ~94.5% refpectively. The limits of detection in honey b.# HPLC with fluorescencedetection are 0.01ppm for OTC and TC, and 0.05ppm for CTC and DC respectively. And therecoveries of OTC, TC, CTC and DC from 4 kinds of honey spiked at a level of 0,Sppm byHPLC with UV detection are 78.3 ~97.0%, 76,8 ~86.8%, 89.6 ~99.5%, 91,1 ~93,8% respectively.The Ihuts of detection in honey by HPLC with UV detection are 0.01ppm for OTC and TC,and 0.02ppm for CTC and DC respectively The residual TCs(OTC, TC, CTC, DC) were notde7ected in the 169 samples of honey(domestic honey 140, imported honey 29) by two HPLCmethods.