두경부종양 세포주에서 Ku 단백질 발현 정도에 따른 방사선 민감도

이상욱(Sang-wook Lee),유은실(Eunsil Yu),이소령(So-Lyoung Yi),손세희(Se-Hee Son),김종훈(Jong Hoon Kim),안승도(Seung Do Ahn),신성수(Seong Soo Shin),최은경(Eun Kyung Choi) 대한방사선종양학회 2004 Radiation Oncology Journal Vol.22 No.3

목 적: DNA-dependent protein kinase (DNA-PK)는 serine/threonine kinase로 470 kDa의 catalytic subunit (DNA- PKcs)와 각각 70 kDa과 86 kDa의 무게를 갖는 Ku 70, Ku 80 단백질로 구성된다. 이 DNA-PK는 방사선에 의해 DNA의 두 가닥이 동시에 절단되는 경우 DNA 손상 복구에 핵심적 역할을 하는 것으로 알려져 있다. 본 연구에서 Ku 발현 과 이온화방사선에 민감도와의 상관 관계를 알아보고자 하였다. 대상 및 방법: 예비실험으로 두경두종양 기원의 세포주 9개에 대한 방사선에 대한 민감도실험을 한 결과 AMC-HN3이 방사선에 가장 민감하였고, AMC-HN9이 방사선에 가장 저항성을 보여 2개의 세포주에 대한 Ku70/80 의 발현을 Western blot과 면역형광 염색을 시행하여 방사선의 반응도와의 상관관계를 알아보았다. 결 과: 방사선에 저항성을 보이는 AMC-HN9에서 Ku80 발현이 높았고 방사선에 민감한 AMC-HN3에서 세포자멸사가 더 많이 일어남을 관찰할 수 있었다. 결 론: Ku80 단백질 발현이 높은 세포는 방사선에 대한 DNA 손상 복구가 많이 되어 방사선에 대한 내성을 보이는 것으로 생각되었다. Purpose: DNA-dependent protein kinase (DNA-PK) is a serine/threonine kinase consisting of a 470 kDa catalytic subunit (DNA-PKcs) and a heterodimeric regulatory complex, called Ku, which is composed of 70 kDa (Ku 70) and 86 kDa (Ku 80) proteins. The DNA-PK has been shown to play a pivotal role in rejoining DNA double-strand-breaks (dsb) in mammalian cells. The purpose of this study is to examine the relationship between the level of Ku expression and radiation sensitivity. Methods and Materials: Nine head and neck, cancer cell lines showed various intrinsic radiation sensitivities. Among the nine, AMC-HN-3 cell was the most sensitive for X-ray irradiation and AMC-HN-9 cell was the most resistance. The most sensitive and resistant cell lines were selected and the test sensitivity of radiation and expression of Ku were measured. Radiation sensitivity was obtained by colony forming assay and Ku protein expression using Western blot analysis. Results: Ku80 increased expression by radiation, wheres Ku70 did not. Overexpression of Ku80 protein increased radiation resistance in AMC-HN9 cell line. There was a correlation between Ku80 expression and radiation resistance. Ku80 was shown to play an important role in radiation damage response. Conclusion: Induction of Ku80 expression had an important role in DNA damage repair by radiation. Ku80 expression may be an effective predictive assay of radiosensitivity on head and neck cancer.

이번 호의 간 병리 : 자가면역 간염

김현정 ( Kim Hyeonjeong ),유은실 ( Yu Eunsil ) 대한간학회 2003 Clinical and Molecular Hepatology(대한간학회지) Vol.9 No.4

비인강암에서 방사선치료의 예측인자로써 Ku70 발현의 의의

송시열(Si Yeol Song),이상욱(Sang-wook Lee),유은실(Eunsil Yu),조경자(Kyung-Ja Cho),박진홍(Jin-hong Park),김상윤(Sang Yoon Kim),남순열(Soon Yuhl Nam),이봉재(Bong-Jae Lee),김성배(Sung Bae Kim),최승호(Seung-Ho Choi),안승도(Seung Do Ahn) 대한방사선종양학회 2005 Radiation Oncology Journal Vol.23 No.1

목 적: 방사선치료를 시행 받은 비인강암 환자에서 Ku70과 DNA-PKcs의 발현 정도가 국소제어율과 치료 실패 양 상에 미치는 영향을 알아보고자 하였다. 대상 및 방법: 1995년 6월부터 2001년 12월까지 서울아산병원에서 방사선치료 단독 또는 동시항암화학-방사선치료를 시행 받은 66명의 원격전이가 없는 비인강암 환자를 후향적으로 분석하였다. 진단 시 얻은 조직검사 표본에서 Ku70과 DNA-PKcs의 면역활성도를 분류하였는데, 그 기준은 면역활성도가 50% 이상 Ku (+)과, 50% 미만 Ku(-) 이었다. Ku70과 DNA-PKcs의 면역활성도와 방사선치료에 대한 종양의 반응 정도와 재발양상과의 상관관계를 알아보았으며, 비인강암의 국소제어율에 영향을 미치는 예후인자를 알아보기 위하여 단변량분석을 시행하였다. 결 과: Ku (-) 환자에서 Ku (+) 환자보다 높은 5년 국소제어율(85% vs. 42%, p=0.042)을 보였으나, 원격전이율은 두 집단간에 차이를 보이지 않았다(78% vs. 82%, p=0.8672). 단변량분석결과 Ku70의 과발현이 기존에 알려진 비인 강암의 예후인자보다 우위에 있음을 알 수 있었다. 국소재발을 보였던 22명의 환자 중 18명의 환자에서 Ku70 (+) 으로 관찰되었다. DNA-PKcs의 발현 정도는 방사선치료 결과에 영향을 미치지 않는 것으로 나타났다. 결 론: Ku70은 원격전이가 없는 비인강암 환자에서 방사선치료 단독 또는 동시항암화학-방사선치료 시행 이후 종양의 반응과 국소제어율을 예측할 수 있는 분자생물학적인 예측인자로 사용될 수 있을 것으로 생각한다. Purpose: The objective of this study was to determine whether the expressions of the two components of DNA-dependent protein kinase, Ku70 and DNA-PKcs, influence the response to radiotherapy (RT) and outcome of treatment of non-disseminated nasopharyngeal carcinoma (NPC) in patients who received definitive RT. Materials and Methods: Sixty-six patients with NPC who were treated with radiotherapy alone or with concurrent chemotherapy between June 1995 and December 2001 were divided into groups based on the levels of immunoreactivity for Ku70 and DNA-PKcs in pretreatment biopsy specimens. The over-expression of Ku70 or DNA-PKcs groups included patients whose biopsy specimens showed at least 50% immunopositive tumor cells; patients in which less than 50% of the tumor cells in the biopsy tissues were immunopositive were placed in the low Ku70 and DNA-PKcs groups. The immunoreactivities for Ku70 and DNA-PKcs were retrospectively compared with the sensitivity of the tumor to radiation and the patterns of therapy failure. Univariate analyses were performed to determine the prognostic factors that influenced locoregional control of NPC. Results: The five-year locoregional control rate was significantly higher in the low Ku70 group (Ku(-)) (85%) than in the high Ku70 group (Ku(+)) (42%) (p=0.0042). However, there were no differences in the metastases- free survival rates between the two groups (Ku70 (+), 82%; Ku70 (-), 78%; p=0.8672). Univariate analysis indicated that the over-expression of Ku70 surpassed other well-known predictive clinocopathologic parameters as an independent prognostic factor for locoregional control. Eighteen of 22 patients who had locoregional recurrences of the tumor displayed an over-expression of Ku70. No significant association was found between the level of DNA-PKcs expression and the clinical outcome. Conclusion: Our data suggest that the level of Ku70 expression can be used as a molecular marker to predict the response to RT and the locoregional control after RT and concurrent chemotherapy in patients with non-disseminated NPC.

사례보고 : 중증 간질환을 동반한 적혈구조혈성 포르피린증 환자에서의 cholestyramine 치료: 사례보고

유동준 ( Dong Jun Yoo ),이한주 ( Han Chu Lee ),유은실 ( Eunsil Yu ),진영주 ( Young Joo Jin ),심주현 ( Ju Hyun Shim ),김강모 ( Kang Mo Kim ),임영석 ( Young Suk Lim ),정영화 ( Young Hwa Chung ),이영상 ( Yung Sang Lee ),서동진 ( Dong 대한간학회 2010 Clinical and Molecular Hepatology(대한간학회지) Vol.16 No.1

EPP는 ferrochelatase의 결핍이나 활성 감소로 프로토포르피린이 적혈구, 간, 혈장 등에 축적되는 유전질환이다 축적된 프로토포르피린은 EPP의 주된 임상적 특징인 광과민 증상과 간질환을 일으키게 된다. EPP로 인한 간질환에 대해 다양한 치료법이 개발되고 있으나, 효과적인 치료 방법은 아직 없다. 그 중 cholestyramine resion의 유용성에 대한 보고는 드물며, 다른 치료와 같이 대부분 사용되었다. 저자들은 cholestyramine resion의 단독치료로 성공적인 결과를 나타낸 사례를 보고하는 바이다. 향후 EPP로 인한 간질환의 증상 및 예후 개선을 위해서, 이번 사례를 포함한 더 많은 연구가 필요하다. Erythropoietic protoporphyria (EPP) is a rare disorder of heme biosynthesis caused by mutations in the gene encoding the enzyme ferrochelatase. In EPP, deficient ferrochelatase activity leads to the excessive production and biliary excretion of protoporphyrin (PP). The major clinical features of EPP are photosensitivity and hepatobiliary disease that may progress to severe liver disease, that are caused by the toxicity of PP. EPP-related liver disease has been treated medically or surgically including liver transplantation. We described a 20-year-old male with severe liver disease who was diagnosed with EPP based on clinical and laboratory findings. He was treated with cholestyramine resin. Six months after the treatment, he was doing well without any abdominal pain or photosensitivity. (Korean J Hepatol 2010;16:83-88)

위 점막연관림프조직 림프종에서 헬리코박터 치료 후 발견된 림프구 위염 -2 증례 보고-

송동은 ( Dong Eun Song ),김정선 ( Jung-Sun Kim ),허주령 ( Joo Ryung Huh ),최진 ( Jene Choi ),장세진 ( Se Jin Jang ),유은실 ( Eunsil Yu ) 대한소화기학회 2005 대한소화기학회지 Vol.45 No.5

Both lymphocytic gastritis and gastric mucosa associated lymphoid tissue (MALT) lymphoma are associated with Helicobacter pylori (H. pylori) infection. However, this association has not been fully elucidated. We report two cases of lymphocytic gastritis i

In Vitro Intrinsic Radiosensitivity Of Human Squamous Cell Carcinoma in Primary Culture

최은경(Eun Kyung Choi),양광모(Kwang Mo Yang),이병용(Byong Yong Yi),장혜숙(Hyesook Chang),김상윤(Sang Yoon Kim),남주현(Joo Hyun Nam),유은실(Eunsil Yu),이인철(Inchul Lee) 대한방사선종양학회 1994 Radiation Oncology Journal Vol.12 No.1

종양의 조직학적 형태에 따라 또 같은 조직의 종양에서도 각 환자에 따라 방사선 치료에 대한 반응 정도에는 많은 차이가 관찰된다. 이러한 방사선 감수성을 예측하는 한 방법으로 각 환자에서 떼어낸 종양조직을 일차 배양하여 방사선 조사에 의한 세포 생존 곡선을 구한뒤 2Gy에서의 생존(SF2)을 얻었다. 방사선 치료가 계획된 두경부 종양과 자궁경부암 환자의 종양 조직을 얻어 기계적인 방법으로 미세절편으로 만든 후 collagenase type Ⅳ와 2시간 배양하여 단일 종양세포 혼탁액을 얻었다. Cell adhesive matix로 전처리된 24 well plate에 각 well당 일정수의 세포를 넣어 24시간 배양한뒤 각열에 0, 1, 2, 3, 4, 6Gy의 방사선을 조사하였다. 13일간 배양후 crystal violet으로 염색한뒤 image analysis system을 이용하여 각 well의 광학밀도를 측정하여 세포 생존을 구한다. Linear quadratic model에 의한 생존 곡선을 얻은 뒤 2Gy에서의 생존율을 구하였다. 배양된 세포가 편평상피암세포임을 확인하기 위하여 cytokeratin과 epithelial monoclonal 항체를 이용한 immunocyto-chemical 염색을 하여 형광 현미경으로 관찰하였다. 5명의 두경부종양 환자와 20명의 자궁경부암 환자의 종양조식을 얻어 실험하여 15명(60%) 종양의 2Gy 생존을 얻는데 성공하였다. 10명의 일차 배양 실패의 원인은 단일 종양세포 혼탁액에 종양세포가 너무 적었거나 세포 이식후 배양이 잘 자라지 않은 것으로 판정되었따. 15편평 상피암 세포의 SF2의 평균은 0.55±0.17이었으며 범위는 0.20에서 0.79까지로 같은 편평상피암이라도 각 환자에 따라 SF2 값에 큰 차이를 보이는 것을 알 수 있었다. 이상에서 같은 부위에 생긴 같은 조직 유형의 종양이라도 각 환자마다 SF2 값의 차이가 큰 것으로 보아 방사선 치료의 효과를 예측할 수 있는 한 인자로 SF2 값을 이용할 수 있을 것으로 생각된다. There are a number of reports suggesting that there may be a correlation between the clinical response to radiotherapy in various tumors and the clonogenic survival of cell lines derived from these tumors following exposure to 2 Gy(SF2). Authors conducted this study to determine SF2 for cells in primary culture from surgical specimens. The tumor tissues with squamous cell carcinoma of uterine cervix and head and neck were obtained. The tumor tissues were disaggregated to single cells by incubating with collagenase type Ⅳ for 2hours with constant stirring. Single cell suspensions were inoculated in four 24-well plates precoated with cell adhesive matrix. After 24 hours of incubation at 37°C, rows of four wells were then irradiated, consisting of control set and five other sets each receiving doses of 1, 2, 3, 4, and 6 Gy. After incubating for a total of 13 days, the cultures were stained with crystal violet and survival at each dose was determined by quantitiative image analysis system. To determine whether cell growth was of epithelial origin, immunocytochemical staining with a mixture of cytokeratin and epithelial monoclonal antibodies were performed on cell cultures. During the period of this study, we received 5 squamous cell carcinoma specimens of head and neck and 20 of uterine cervical carcinoma. Of these, 15 yielded enough cells for radiosensitivity testing. This resulted an overall success rate of 60%. The mean SF2 value for 15 tumours was 0.55±0.17 ranging from 0.20 to 0.79. These results indicate that there is a broad range of sensitivities to radiation in same histologic type. So with a large patient population, we plan to determine whether a different SF2 value is associated with tumours that are controlled with radiotherapy than those that are not.

Methionine Adenosyltransferase 1: 간경화 환자에서 단백체학 분석을 통한 조기 간세포암 진단표지자

이주호 ( Joo Ho Lee ),전미정 ( Mi Jung Jun ),심주현 ( Ju Hyun Shim ),송기원 ( Gi-won Song ),탁은영 ( Eunyoung Tak ),오보라 ( Bora Oh ),유은실 ( Eunsil Yu ),최상운 ( Sang-woon Choi ),안지현 ( Jihyun An ),이단비 ( Danbi Lee ),김강모 ( 대한간암학회 2018 대한간암학회지 Vol.18 No.1

Background/Aims: Because there is a lack of effective biomarkers, we aimed to discover proteomic candidate markers for hepatocellular carcinoma (HCC) in cirrhotic patients at the highest-risk of HCC, and to validate the markers. Methods: We collected tumor tissue from 5 cirrhotics with HCC, and from 5 cirrhotics without HCC, who underwent liver resection or transplantation. These tissue samples were analyzed by 2-dimensional difference gel electrophoresis coupled with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), and potential markers were validated at the transcriptional and translational levels. We also performed western blot assays using other blood samples from 10 cirrhotics with HCC and 10 without HCC. Results: Among the 66 distinguishable spots on 2-D gel images, we identified 15 proteins overexpressed more than 1.5 fold in terms of volume ratio in the tumors. Ten of the over-expressed proteins were identified by MALDI-TOF MS; of those, only methionine adenosyltransferase 1 (MAT1), a protein specific for liver, and acyl-CoA dehydrogenase were significantly up-regulated in tumors in further immunoblotting analyses (Ps<0.05). There was no between-pair difference in MAT1 mRNA measured by real-time polymerase chain reaction (P=0.96). However, in western blots of serum samples, distinct MAT1 bands were observed in all 10 HCC patients, but in only 2 of the non-HCC patients. Conclusions: MAT1 is a potential marker for surveillance in cirrhotic patients with and without prior HCC. (J Liver Cancer 2018;18:33-43)

만성 간염의 조직학적 등급 체계 : 대한병리학회 소화기병리연구회 시안을 중심으로 On the Basis of Standardized Guideline Proposed by the Korean Study Group for the Pathology of Digestive Diseases

유은실 대한간학회 2003 Clinical and Molecular Hepatology(대한간학회지) Vol.9 No.1

타액선 종양의 세침흡인 세포학적 소견:조직학적 진단과의 비교

유은실,추광절,김상윤,최기영,공경엽,이인철 울산대학교 의과대학 1993 울산의대학술지 Vol.2 No.1

We reviewed twenty fine needle aspiration biopsies of salivary gland lesions performed between April 1992 and March 1993, which were confirmed by surgical excision. Sixteen cases were benign lesions:10 pleomorphic adenomas, 3 Warthin's tumors, 1 monomorphic adenoma, 1 benign lymphoepithelial lesion and 1 lymphoepithelial cyst. Malignant lesions were four cases:3 mucoepidermoid carcinomas, and 1 malignant mixed tumor. Preoperative fine needle aspiration cytologic findings were compaired with pathologic diagnosis of the surgically resected specimen in all cases. Overall accuracy was 80%. False negativity and positivity were 5% (1 case) and 0%, respectively. Fine needle aspiration biopsy for diagnosis of salivary gland lesion appears to be a simple, safe, and cost saving diagnistic method with relatively high accuracy.

정구 점막선증식증

최미선,이태진,유은실,노재윤 대한병리학회 1999 Journal of Pathology and Translational Medicine Vol.33 No.9