한국형 자궁경부 촬영진 ( New Cervicography ) , 세포검사 그리고 HPV-DNA 검사를 이용한 새로운 자궁경부암 검진 모델 ( Model ) 개발

김승조(SJ Kim),박찬규(CK Park),이효표(HP Lee),남궁성은(SE Namkoong),강순범(SB Kang),서호석(HS Saw),이재관(JK Lee),김수녕(SN Kim),김재원(JW Kim),배석년(SN Bae),김찬주(CJ Kim),이근호(KH Lee),이선영(SY Lee),김인호(IH Kim),이찬(C Lee),이정노(JN Lee) 대한산부인과학회 2001 Obstetrics & Gynecology Science Vol.44 No.5

N/A Objective : The false negative rate of conventional cytology is reported to range from 18 to 45%. It is necessary to develop more effective screening strategies that would ideally be more accurate than conventional cytology. This study is designed to investigate the potential of conventional cytology, cervicography, HPV-DNA testing and their combinations as primary screening techniques for cervical neoplasia. The purpose of this project is to develop the models and guidelines for screening tools of cervical cancer of the uterus by evaluating sensitivity, specificity and cost-effectiveness of all the screening methods. Study design : We conducted a screening of random sample of women who visited the department of OB-Gyn. of five different major university hospitals such as Catholic University Hospital, Korea University Hospital, Seoul National University Hospital, Yeonsei University Hospital and Pochon CHA University Bundang CHA Hospital, from May 1, 1996 to April 30, 1999. In a cohort women with an age range of 20 to 70 who underwent routine cytologic screening at Catholic University, Hospital Bundang CHA General Hospital and local clinics from April 1, 1996 to Decomber 31, 2000. cervicography (n=417,125) and testing for HPV-DNA (n=1,347) by the hybrid capture assay were studied for the ability of the cervical cancer screeuing. A new cervicography system with Kim's classification which was developed by Prof. Kim Seung Jo as an adjunctive method for the cervical cancer screening was applied in this cohort study. And then, accuracy, effectivencess, cost-effectiveness of the single or combined screening method were analysed. Result : Sensitivity and specificity of Pap smear were ranged from 55.6% to 83.1% and 72.8% to 88.3% respectively. The combination of Pap smear and cervicography had sensitivity from 89.7% to 98.6%, specificity from 68.2% to 93.2%. With combination of Pap smear, HPV DNA test and cervicography, sensitivity became also highest accuracy among all screening methods from 92.8% to 98.8%. Considering medical charges for diagnosis and social cost occurred by false positive and false negative results, the most cost-effective diagnostic modality was thought to be the combination of Pap smear and cervicography(91,433 won). In patients who were diagnosed as LSIL, colposcopic examination confirmed progression to high grade intraepithelial lesion(HSIL) in 10.7% patients, persistence of LSIL in 55% patients during the 3 year follow-up period. Accuracy of the screening for cervical cancer and CIN can significantly be improved by cytology with new cervicography rather than cytology alone. We concluded that cervicography can be important adjunctive tests for cervical cytology, improving the effectiveness of cervical screening by allowing a more sensitive detection of cervical neoplasia.

난관복원술의 성공율을 높이는 제반여건의 고찰

김종덕,두재균,류철희,차경연,양재이 대한산부인과학회 1991 Obstetrics & Gynecology Science Vol.34 No.6

난관복원술의 성공율을 높이는 제반 여건을 알아보기 위하여 1982년 3월부터 1990년 2월까지 만 8년 동안 전북대학교병원 산부인과 불임크리닉에 내원하여서 미세수술을 이용한 난과복원술을 시행받은 총 116명중 6개월이상이 추적조사가 가능하였던 106명의 예를 분석하여 다음과 같은 결론을 얻었다. 1. 난관복원술후 임신성공율은 추적조사가 가능하였던 106명중 90명(84.9%)이었다. 2. 난관복원술의 동기는 자녀사망이 전체 평균 65.1로 가장 많았으나 1987년이후부터는 재혼(24.6%)과 심경의 변화(20.0%) 등이 증가하여 서구화되는 경향을 보여주고 있었다. 3. 난관복원술후 임신군의 평균연령은 28.5세, 비임신군은 30.3세이었다(p$lt;0.05). 4. 불임수술후 난관복원술까지의 평균기간은 임신군에서 35.5개월, 비임신군에서 42.4개월로 임신군에서 그 기간이 짧았다(p$lt;0.05). 5. 임신성공율에 따른 복원술후 난관 길이는 임신군 8.3 cm, 비임신군 6.8 cm이었다(p$lt;0.05). 6. 불임수술 방법에 따른 결과는 Falope ring군이 난관의 길이도 가장 길고 임신성공율도 91.7%로 가장 높았다(p$lt;0.05). 7. 난관복원술 부위에 따른 임신성공율은 팽대부-팽대부, 자궁각부-팽대부, 협부-협부, 협부-팽대부순이었다(p$lt;0.05). 8. 난관복원술후의 임신하기까지의 기간은 임신군에서 1년이내가 95.5%로 가장 높았으며 2년이후에는 1예도 없었다. 9. 난관복원술후 난관임신 발생율은 1명(0.9%)이었다. 10. 본 교실에서 고안하여 사용하고 있는 Kim`s clamp와 Doo`s microirrigator 등은 난관성형술에 있어서 아주 유용한 기구로 사료되었다. For evaluation of factors that influence the success rate of tubal reversal, 116 cases of microsurgical reversal were investigated, which were performed at the infertility clinic in the Department of Obstetrics and Gynecology, Chonbuk National University Hospital from March 1982 to February 1990. 106 cases were able to follow up more than 6 months among these 116 cases of tubal reversal patients. The results were as follows; 1. The pregnancy rate was 84.9%(90 patients) in 106 cases of tubal reversal. 2. The most common incentive was the loss of their children(75.5%), but since 1987 remarriage(24.6%) and change of marital attitude(20.0%) were increasing steadily. 3. After tubal reversal, the mean age of pregnant patients was 28.5 years and that of non-pregnant patients was 30.3 years(p$lt;0.05). 4. The time interval between the sterilization and the tubal reversal was 35.5 months in pregnancy groups, 42.4 months in non-pregnancy groups(p$lt;0.05). 5. The mean length of reconstructed tube was 8.3 cm in pregnancy patients, 6.8 cm in non-pregnant patients(p$lt;0.05). 6. Referring to the sterilization types, the pregnancy rate was higher and the length of reconstructed tube was longer in those who underwent tubal ligation with Falope ring. 7. Referring to the site of anastomosis, the pregnancy rate was higher in ampulloampullar, cornu-ampullary, isthmo-isthmic anastomosis than other group(p$lt;0.05). 8. The pregnancy rate within the first year after tubal reversal was 95.5%, but no case after 2 years was noted. 9. Postoperative tubal pregnancy developed in one case(0.9%) after tubal reversal. 10. Kim`s clamp and Doo`s microirrigator were considered to be valuable instruments in tubal surgery.

인간 상피성 자궁경부암 세포주 (CUMC-6) 의 확립 및 특징화

김승조,정재근,김진우,남궁성은,박태철,김흥기,김수평,김사진,김태웅,김재훈 대한산부인과학회 1996 Obstetrics & Gynecology Science Vol.39 No.1

본 연구진은 31세된 한국여성의 자궁경부암 종양조직을 채취한 후 세포배양을 통하여 자궁경부의 편평상피암 세포주(CUMC-6)를 새로이 확립하여 시험관내 성장특성, 세포의 형태학적 특성, 세포의 초미세 구조, 생체내 종양형성 능력, 동위효소 발현, 염색체 특성, 호르몬 및 태반단백 분비, 종양표지물질분비, 스테로이드 호르몬 수용체 여부, 인체 백혈구 항원의 규명, 인유두종 바이러스(HPV) 감염,c-myc및 L-myc 발암유전자들의 변이, 및 p53 종양 억제 유전자의 변이여부를 검색하여 다음과 같은 결과를 얻었다. 1. 자궁경부암 세포주는 현재 61개월간 배양중에 있으며 지속적인 성장을 보였고 현재까지 연속적으로 300계대까지 수행되었다. 2. 배양중인 종양세포의 성장특성을 분석한 결과 population doubling time은 약 36시간이었으며 plating efficiency는 32%로 나타났다. 3. 배양세포주의 형태학적 특성을 규명하고자 위상차 현미경 소견, H & E 염색, Papanicolaou`s 염색 및 전자현미경적 방법 등을 이용한 결과 인간 상피세포 유래 종양임을 확인할 수 있다. 4. 배양세포주를 누드마우스에 이종이식시켜 종양의 형태학적 변화를 분석한 결과 형성된 종양조직은 본래의 조직소견과 유사하였으나 분화정도는 나빴다. 또한, 종양형성 능력을 파악한 결과 2주 이내에 96%의 높은 종양형성 능력을 보였다. 5. 자궁암 세포주의 glucose-6-phosphate dehydrogenase 동위효소 분석시 type B pattern을 보여 기존의 HeLa 세포주와의 실험실내 오염이 아닌것으로 판명되었다. 6. CUMC-6 종양세포주들의 염색체 분석시 대부분의 세포들은 이배수성(2n-46)을 나타내었으며 약 20%의 세포들에서 7번 염색체 하나가 더 많은 삼배수성(trisomy)을 보였다. 7. 태반단백, 호르몬 및 종양표지물질 분비여부를 검색한 결과 태반단백으로서는 B-HCG 분비를 확인하였고, 또한 종양표지물질로서는 AFP분비를 확인하였다. 8. 종양세포주의 에스트로겐 및 프로게스테론 수용체 존재여부를 분석한 결과 각각 음성으로 판정되었다. 9. 종양세포주의 인체 백혈구항원(human leukocyte antigen) 검색 결과 HLA class II phenotype 들은 DR12와 DQw3로 나타났으며, HLA class II allele specificity들은 DRB1*1202, DQA1*0601 , 및DQB1*03*들로 판명되었다. 10. 배양된 CUMC-6 세포주에서 HPV 감염여부를 검색하기 위하여 polymerase chain reaction(PCR) 을 실시한 결과 HVP 16과 HVP 18이 동시에 감염되어 있는것을 확인할 수 있었다. 11. CUMC-6 세포주에서 c-myc 유전자는 재배열 및 증폭소견들을 보여주었으나 L-myc 유전자의 재배열 및 증폭 소견들은 발견되지 않았다. 12. Polymerase chain reaction-signle strand conformation polymorphism(PCR-SSCP)방법으로 배양된 CUMC-6 세포주에서 p53 종양억제 유전자의 변이는 관찰되지 않았다. 이와같이 특징화 과정이 완료된 CUMC-6 자궁경부 상피암 세포주는 향후 한국 여성의 자궁경부암 치료와 생물학적 연구에 적절한 실험 모델이 될것으로 사료되며 자궁경부 상피암의 발생기전에 염색체 1번의 이상, c-myc 발암유전자의 변이, 및 HVP 감염등이 복합적으로 관여하는 것을 파악할 수 있었다. A new cell line, CUMC-6, has been derived from an invasive non-keratinizing squamous cell carcinoma of the uterine cervix in a 31-year-old patient. It has been maintained in long term culture for 61 months, and passaged over 300 times. The results were as follows; 1. Monolayer-cultured cells were polygonal in shape, showing a pavement-like arrangement and a tendency to pile up without contact inhibition. 2. The epithelial nature of the cultured CUMC-6 cells was confirmed by transmission electron microscopy which demonstrated the presence of desmosomes and tonofilaments. 3. The sbucutaneous injection of cultured cells into nude mice gave rise to fast growing tumor. The transplanted tumor showed similar histological features, but poor differentiation than the original tumor. 4. Repeated chromosome analysis revealed that most metaphases of this cell line contained diploid number of chromosomes. The consistently observed structural abnormality in this cell line was elongation of short arm of chromosome 1. The G-banded pattern of this chromosome was inv dup (1)(1pter-$gt;1p34:: 1p21-$gt;34:: 1p34-$gt;1qter). 5. The results of isozyme analyses were distinct from the HeLa cell line. 6. Cultured CUMC-6 cells produced human chorionic gonadotropin B-subunit(B-HCG),and a-fetoprotein(AFP). 7. Cytosol estrogen and progesterone receptors were not measured in this cell line. 8. Human leukocyte antigen(HLA) typing of CUMC-6 cells indicated the presence of DR12 and DQw3. 9. Analysis of the DNA extracted from the CUMC-6 cells showed the presence of human papillomavirus(HPV) type 16 and 18 DNAs. 10. Southern blotting technique revealed ampification and rearrangement of oncogene c-myc and no amplification of L-myc. 11. Using the polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP) technique, we have screened CUMC-6 cells for p53 mutation in exons 4 to 9. No mobility shift was observed in this cell cline. This cell line will be a valuable tool in studying the in vitro and in vivo properties of human cervical carcinoma.

자궁경부암 세포주들에서 리포좀을 이용한 종양억제 유전자 [pRcCMVp53/liposome]의 직접 이입법에 의한 세포 성장 억제 효과

김승조,이헌영,남궁성은,김수평,이준모,안웅식,김종국,박순희,박용석,김진철,한유진,양재명,신봉영,서상용,김동재 대한산부인과학회 1999 Obstetrics & Gynecology Science Vol.42 No.4

목적: 자궁경부암 유전자 치료를 위하여 자궁경부암 세포주들에 리포좀을 이용한 종양억제 유전자[wild-type p53 gene]의 직접 이입을 통하여 세포성장 억제효과를 보고자 하였다. 방법: 자궁암세포주들 [CaSki, SiHa, HeLa, HeLaS3, C33A, HT3]을 12 well에 104, 96 well에 5×103의 수로 triplicate를 형성한 뒤 대조군, pRcCMVp53, lipofectin, pRcCMV/lipofectin, pRcCMVp53/lipofetin이 함유되있는 것 을 각기 12 well에 20 ㎕씩 96 well에는 10 ㎕씩을 투여한 다음 7일간 배양하였다. 세포수 측정, ELISA [neutral red] assay, MTT등을 이용하여 세포성장과 cell viability를 측정하고 이를 one-way ANOVA F-test, multiple comparison Tukey method에 의해 날짜별 서로 군의 차이를 비교 하였다. 결과: p53 유전자에 의한 자궁경부암 세포주의 성장억제를 확인하기 위하여 세포 수 측정, ELISA assay, MTT assay를 시행하였으며 각각의 assay에 대한 측정 마지막날인 6일째를 기준으로 정상대조군 세포와 비교한 결과 세 포수 측정에서는 CaSki[89%], SiHa[59.2%], HeLa[86%], HeLaS3[78%], C33A[91%] 그리고 HT3[74%] 성장억제를 나타냈다. 또한 Neutral red를 이용한 세포 viability assay 에서는 CaSki[70%], SiHa[93%], HeLa[73%], HeLaS3[50%], C33A[67%] 그리고 HT3[52%]에서 는 52%의 세포감소를 보여주었다. MTT assay를 통한 각각의 자궁경부암 세포주의 세포성장억제 결과는 CaSki[38%], SiHa[72%], HeLa[53%], HeLaS3[28%], C33A[33%]그리고 HT3[53%]였다. 결론: 자궁경부암 세포주에 직접 이입법을 통한 유전자 전달 후 세포성장 억제는 매우 유익하게 감소되었으며 이와같은 결과는 향후 유전자 직접 이입법에 의한 자궁경부암 환자에 있어서 유전자 치료를 실시할 수 있는 새로운 장으 로 생각이 되어진다. Purpose: We investigated cell growth inhibitory effect of the wild type p53 gene into the cervical cancer cells via the recombinant p53 plasmid, pRcCMVp53 with lipofectin. Method: Inhibition of the growth of cervical cells as determined by a cell count assay. The cells were inoculated at density of 104 cells/well in each 12 well plate, 24 hrs before infection. At each indicated point, cells in three wells on each well plate were trypsinized and counted. The mean cell counts of triplicated well after transfection at day 1~6 was checked. Inhibition of the growth of cervical cells were checked by ELISA assay, MTT assay. The cells inoculated at densities of 5×103/well in each 96 plate 24 hrs before infection. At each indicated point cells of three wells were transfected for 6 days, harvested and counted by liquid scintillation counter the mean cpm per triplicate wells were plated for ELISA and MTT assay. One-way ANOVA F-test and multiple comparison Tukey method was used for statistical analysis. Result: Inhibition of the growth of cervical cells in cell count showed CaSki[89%], SiHa[59.2%], HeLa[86%], HeLaS3[78%], C33A[91%] and HT3[74%]. ELISA assay showed CaSki[70%], SiHa[53%], HeLa[73%], HeLaS3[50%], C33A[67%] and HT3[73%]. MTT assay showed HeLa[33%], HeLaS3[28%], SiHa[75%], CaSki[38%], C33A[33%] and HT3[53%]. Conclusion: These results indicate that transfection of cervical cancer cells with the wild type p53 gene via pRcCMVp53 with lipofectin is a potential novel approach to the gene therapy of cervical cancer.

Anti-Diabetic Effects of DY-101267, A Potent PPARs Agonist

Lee CH,Kim MK,Shin CY,Chae YN,Son MH,Kim JK,Min JP,Moon HS,Kim HJ,Bae MH,Kim SH,Lee JG,Lim JI 대한약학회 2004 大韓藥學會 總會 및 學術大會 Vol.2004 No.2

Krukenberg 종양 1 예

고재근(JK Ko),김현호(HH Kim),신재경(JK Shin),김경호(KH Kim),김영래(YR Kim) 대한산부인과학회 1988 Obstetrics & Gynecology Science Vol.31 No.6

위의 원발암종이 난소에 전이된 것으로 보이는 Krukenberg 종양 1예를 경험하였기에 문헌적 고찰과 함께 보고하는 바이다. Krukenberg tumor is comparatively rare in metastatic ovarian tumors . We have experienced one case of Krukenberg tumor which are presented with a brief review of literatures

금강초롱꽃의 자생지 식생 및 생육조사

임학태(Lim HT),김종화(Kim JH),김정기(Kim JK),이우철(Lee WT),유기억(Yoo KO),김원배(Kim WB),김병현(Kim BH),김정간(Kim JK) 한국원예학회 1993 한국원예학회 학술발표요지 Vol.11 No.2

導入 機器의 效率的 活用을 위한 敎授-學習 敎材의 開發 : 8 個 學科 共同 硏究

李昌甲,孟琦錫,李載元,朴萬植,廷吉煥,金鍾悟,朴景采,李英浩,李東周,邕章祐,柳盛淵,趙澤東,姜理錫,李殷雄,李興浩,金健中,韓相玉,金良模,張錫明,朴俊灝,洪鳳植,金太均,金再韓,宋海永,盧載星 충남대학교 공업교육연구소 1982 論文集 Vol.5 No.1

In this study, through the combined effort and collaboration of 8 engineering education departments, teaching-learning processes and models were reviewed, and printed and bound student manuals developed on 120 different tasks and subjects. Audio-visual supportive material, already in use in laboratories, was added to form a complete teaching-learning package. In addition to promoting the maximum use of the newly acquired equipment, this study will also contribute to the development of laboratory practice, the in-service training of technical high schools teachers and help in the development of their own instructional program.

Transforming Growth Factor- 1에 의한 자궁경부암 세포주들의 세포성장억제 기전

김승조,이헌영,정재근,김진우,김미란,남궁성은,김수평,이준모,한구택 대한산부인과학회 1998 Obstetrics & Gynecology Science Vol.41 No.4

TGF- 1은 대부분의 상피세포를 포함한 여러 종류의 정상세포들에서 세포성장 억제 효과를 나타내는 것으로 알려져 있으나 자궁경부암 세포의 성장에 대한 TGF- 1의 분자생물학적인 기전은 아직 확실하게 밝혀져 있지 않다. 저자들은 본 교실에서 확립한 자궁경부암 세포주인 CUMC-3와 CUMC-6를 이용하여 자궁경부암 세포주의 성장에 미치는 TGF- 1의 영향을 알아보기 위하여 본 연구를 시행하여 다음과 같은 결과를 얻었다. 1) TGF- 1을 0.1, 1 및 10 ng/ml 농도로 24시간 동 안 처리하였을 때 MTT 법을 이용하여 측정된 자궁 경부암 세포주들의 성장은 농도에 비례하여 억제되 었다. 2) TGF- 1의 종양세포성장 억제 효과는 아폽토시 스의 유도에 의한 것으로 생각되었다. 3) Northern blot 상 TGF- 1을 투여 후 세포성장을 조절하는 유전자인 c-myc mRNA의 발현이 두 세포 주들에서 억제되었으며 c-myc에 대한 억제 효과는 CUMC-6 자궁경부암 세포주에서 훨씬 강력하였다. 4) TGF- 1 투여 후 p27Kip1 단백의 농도가 증가되 었음이 western blot 상 두 세포주에서 관찰되었다. 이러한 결과들로 보아 TGF- 1이 c-myc 유전자의 발현억제, p27Kip1 단백의 과도한 발현과 아폽토시스 의 유도로 자궁경부암 세포주의 성장을 억제하였으 며, CUMC-3와 CUMC-6 자궁경부암 세포주가 TGF- 1의 세포성장 억제기전을 밝히는 데 좋은 모형이 될 것으로 생각된다. Transforming growth factor- 1 (TGF- 1) is known to be a potent growth inhibitor of many cell types including most epithelial cells. However, the molecular mechanism of TGF- 1 action on cell growth of cervical carcinoma has not yet been elucidated. We assessed the effect of TGF- 1 on the cervical carcinoma cell lines, CUMC-3 and CUMC-6, which we estabilished in our laboratory. The results of this study were as follows: 1) TGF- 1 inhibited the growth dose-dependently at the concentration of 0.1-10 ng/ml in both cell lines. 2) DNA fragmentation and electron microscopic findings showed that TGF- 1 induced apoptosis in both cell lines 3) On northern bolt analysis, c-myc mRNA expression was suppressed by TGF- 1 in both cell lines. The inhibition of c-myc expression by TGF- 1 was more prominent in CUMC-6 than in CUMC-3. 4) Western blot analysis showed that the level of p27Kip1 protein was increased after TGF- 1 treatment in both cell lines. These results suggest that TGF- 1 induce apoptosis in two cervical carcinoma cell lines, CUMC-3 and CUMC-6, by the mechanisms involving down-regulation of c-myc gene and over-expression of p27Kip1 protein. We also suggest that CUMC-3 and CUMC-6 may be a good model for study of the complex mechanism of TGF- 1.

Krukenberg 종양 3 예

김동진,신영관,이석환,김태웅,김준길,윤미림 대한산부인과학회 1993 Obstetrics & Gynecology Science Vol.36 No.10

저자들은 Krukenberg종양 3 예를 치료했으나 모두 수술후 20 개월이내에 사망하였다. Krukenberg tumor is comparatively rare in metastatic ovarian tumor and is almost metastasized from gastrointestinal tract. Early diagnosis and treatment is difficult. The definite diagnosis would be pathological diagnosis with surgical specimen. We experienced 3 cases of krukenberg tumor on the ovary which were metastasized from the adenocarcinoma of stomach. The first died 24months later after subtotal hysterectomy with B.S.O. The second died 9 months later after B.S.O and the third died 16 months after T.A.H. with B.S.O. All were received postoperative adjunctive chemotheraphy. We present these cases with the brief review of literature.