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.

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.

Surface Modification of Titanium Implant by Anodic Oxidation Treatment and Bisphosphate Immobilization

Kim WG,Lee SJ,Soh YJ,Lee MH,Kim HS,Kim BI,Bae TS 대한치과재료학회 2011 대한치과기재학회 학술대회 Vol.2011 No.5

쌍태임신중 일측태아의 사망 1 예

김영덕,김용욱,김휴,이석주 대한산부인과학회 1997 Obstetrics & Gynecology Science Vol.40 No.8

The antepartum death of one fetus in twin pregnancy is a rare obstetric complcation. If one fetus in twin pregnancy died in uterus remote from term and another fetus did not, the dead fetus will be compressed between the uterine wall and the membrane of living fetus, becomes a fetus compressus or fetus papyraceous. The primary maternal threat when the fetuses undelivered is disseminated intravascular coagulation. Concern for the surviving fetus after death of its co-twin is balanced between the risks of preterm birth and those involving exposure to events in utero that may threaten its well-being. Recently we experienced a case of twin pregnancy complicated by the death of one fetus, we report this case with concerned literatures.

완전포상기태 예후인자로서 접합체형의 임상적 의의

김승조,배석년,신진웅,김찬주,김재훈,이상형,김재동,윤수철 대한산부인과학회 1997 Obstetrics & Gynecology Science Vol.40 No.10

To evaluate the possible correlation between the origin of complete hydatidiform mole(CHM) and subsequent persistent gestational trophoblastic tumor(GTT) after molar evacuation, we have studied genetic origin patterns against conventional clinical parameters -patient`s age, gestational age, uterine size for gestational age, serum β-hCG levels before molar evacuation- in 69 patients with CHM. In our study, each of large uterine size for gesta-tional age, serum β-hCG levels before molar evacuation, and genetic origin of CHM had a prognostic significance of subsequent persistent GTT. However, each of gestational age and patient`s age is not a good prognostic indicator for subsequent persistent GTT. Among the patients with persistent GTT, there are no differences in clinical parameters- patient`s age, gestational age, tumor age(the interval between evacuation of CHM and initiation of chemotherapy), serum β-hCG levels before molar evacuation and before initiation of chemotherapy- according to the origin of CHM. There are no differences in the analysis of sex-chromosome and variable number tandem repeat sequence YNZ22 and APOB gene in the extracted DNA from frozen tissues and paraffin blocks and from EDTA treated peripheral blood and dried blood specimen on Wartman paper. It is suggested that analysis of sex-chromosome and polymorphism of YNZ22 and APOB gene from the extracted DNA of paraffin block and dried blood specimen on Wartman paper is the valauble experiment to evaluate the origin and the classification of hydatidiform mole and seems to be the sensitive molecular genetic method in predicting subsequent persistent GTT.

자궁경부암 세포주들에서 Transforming Growth Factor-β1의 세포성장 및 c-myc암유전자 발현 억제효과

김승조,이헌영,김진우,김미란,남궁성은,박태철,김태웅,김재훈 대한산부인과학회 1997 Obstetrics & Gynecology Science Vol.40 No.1

Transforming growth factor-β1(TGF-β1) is known to be a potent growth inhibitor for many cell types, including most epithelial cells. In skin keratinocytes, TGF-β1 has been shown to inhibit growth and to rapidly reduce c-myc expression. However, the molecular mechanism of TGF-β1 action on cell growth of cervical carcinoma has not yet been elucidated. We thus assessed the effect of TGF-β1 on the growth of cervical carcinoma cell lines. Two cervical squamous carcinoma cell lines were incubated with varying concentration of TGF-β 1, and growth inhibition was evaluated with tetrazolium-based colorimetric assay. After culturing in concentrations of 0.1~10 ng/ml in both cell lines. Northern blot analysis revealed c-myc mRNA expression was suppressed by 10 ng/ml of TGF-β 1 following 6-hour of treatment in both cell lines. These results suggest that TGF-β1 inhibits the growth of cervical carcinoma cells by suppressing c-myc oncogene expression.

자궁경부암 근치술후의 성생활의 변화에 관한 연구

김승조,정재근,남궁성은,지용헌,김대훈,김도강,이원 대한산부인과학회 1991 Obstetrics & Gynecology Science Vol.34 No.1

본 연구는 가톨릭의과대학 부속강남성모병원 산부인과 환자중 경부암 근치술을 받은 53명을 실험군으로 하고 단순자궁절제술을 받은 51명을 대조군으로 하여 양군의 성생활과 관련된 신체적 정신적 스트레스에 대한 면담조사 및 Hamilton의 우울척도에 의한 우울성향을 조사하여 다음과 같은 결론을 얻었다. 1. 경부암 근치술을 받은 환자는 단순자궁절제술을 받은 환자에 비하여 교육수준과 경제력이 낮았다. 2. 성생활에 있어서 경부암근치술을 받은 환자는 단순자궁절제술을 받은 환자보다 극치감 감소, 성욕감소, 불감에 있어서 유의한 차이를 보였다. 반면에 남편의 성생활에 있어 발기불능, 조루, 사정불능, 외도에는 차이가 없었으나 성욕감소는 유의하게 증가하였다. 3. 일상생활의 문제점에 있어 경부암근치술을 받은 환자는 단순자궁절제술을 받은 환자보다 남편의 관심상실 우려, 여자로서 자아의 상실에 대한 우려, 남편의 태도변화 우려, 재발우려, 남편에 대한 피해의식이 많았고 같은 환자와의 대화가 많은 도움이 되었다고 한다. 4. 우울성향은 경부암근치술을 받은 환자가 단순자궁절제술을 받은 환자보다 높았다. 5. Hamilton의 우울척도에 있어서 경부암근치술을 받은 환자는 단순자궁절제술을 받은 환자보다 우울감정, 죄의식, 자살, 불면증, 일과 활동, 정신운동 지체, 초조, 정신적 신체적 불안, 일반적 신체증상, 건강 염려증, 무력감, 절망감, 허무감 등이 유의하게 높았다. 그러므로 경부암 근치술 후의 환자관리는 성생활의 변화와 우울감정에 대하여 충고를 해주고 같은 환자와의 접촉을 권하며 가족, 특히 남편의 이해와 협조를 얻어 자기의 생활을 유지해 나갈 수 있도록 도와 주어야 한다. This study was designed to investigate physical and psychological stress and depression after radical hysterectomy by means of clinical interview and Hamilton`s rating scale for depression. The structured interviews of 53 radical hysterectomy patients as experimental group and 51 total hysterectomy patients as control group were performed. The results were as follows. 1. The educational and economical levels of radical hysterectomy patients were lower than that of total hysterectomy patients. 2. The two groups were significantly different in terms of loss of orgasm, loss of sexual desire and frigidity. While there was no significant difference in impotence, premature ejaculation, retarded ejaculation and extramarital sex on the part of the husband except in sexual desire. 3. Significantly more patients of radical hysterectomy thought of loss of husband`s affection, loss of womanliness and change of husband`s attitude as the problem of life than total hysterectomy patients. More radical hysterectomy patients replied that their disease had come from husband and the conversations with the same disease were helpful to them. 4. Depression of radical hysterectomy patients was inclined to be higer than that of total hysterectomy patients. 5. Hamilton`s depression score of radical hysterectomy patient is significantly different from that of total hysterectomy patients in depressive mood, guilt, suicide, insomnia, work and interest, retardation, agitation, psychic and somatic anxiety, somatic general, hypochodriasis, helplessness, hopelessness and worthlessness. In the process of radical hysterectomy patient treating, it is required to advise the patient on change of sexual life and depression, to help the patients to maintain her life through conversations with the same disease and to help the patient to get understanding and cooperation from her husband and members of her family as their attitude has influence on patient`s adaptation.

고위험 자궁경부암환자에서 Quick Cisplatin-VP16 을 이용한 신보조항암요법의 효과

김승조,김진우,배석년,남궁성은,박종섭,이준모,김찬주,김재훈,박동춘,박용규 대한산부인과학회 1997 Obstetrics & Gynecology Science Vol.40 No.8

The potential role and determinants of response to a cisplatin-based regimen of neoadju-vant chemotherapy in women with a histologically confirmed first diagnosis of stage IB-Ⅲ cervical cancer were analyzed. From 1993 to 1996, 92 patients with bulky(designated as more than 3×3 cm2 size) mass were treated with cisplatin 60 mg/m2 and etoposide 100 mg/m2, admi-nistered intravenously at 7 day intervals. Seventy cases of radical hysterectomy with pelvic lymph node dissection and 22 cases of radiation therapy were performed 2 to 3 weeks after chemotherapy. At the end of the cycles, the overall clinical response rate of portio was 83.7 %(34.8 % with a complete response and 48.9 % with a partial response). The older ages, lower stages, and squamous cell types correlated favorably with the clinical response of the portio, but neither with the parametrium nor with the vagina. After the operation, we found the diff-erences in histologic responses, with the following parameters:lymphovascular space invasion, 3 mm below stromal invasion and lymph node metastasis. Theses parameters correlated with the clinical responses, and the down-staging of cases were 70 %. In comparison with radiolog-ical findings of pretreatment and postoperative tissue pathology, we could find a decrease in pelvic LN metastasis. The tumor-free survival rate calculated by the Kaplan-Meier product limit method was 75 % but it was 86.1 % for cases without the occurrence of persistent disease after the completion of the treatments. All patients suffered from nausea and vomiting, but grade 4 toxicity was not detected after the routine use of antiemetics. There were no events that delayed the next step in the treatment or caused difficulty during the operation. The results of this study suggest that the neoadjuvant chemotherapy should be accepted as a routine tool in treating high risk cervical cancer in order to improve the likelihood of favorable outcomes.

Electrophysiological safety of sibutramine HCl

Kim, KS,Park, SJ,Lee, HA,Kim, DK,Kim, EJ Sage Publications 2008 Human & experimental toxicology Vol.27 No.7

<P>Sibutramine is known to induce cardiovascular side effects such as tachycardia, vasodilation, and hypertension. The present study was aimed to examine the effects of sibutramine on action potential of guinea pig papillary muscle, recombinant hERG currents (I<SUB>hERG</SUB>), and inward currents (I<SUB>Na</SUB> and I<SUB>Ca</SUB>) of rat ventricular myocytes. Sibutramine at 30 μg/mL induced a shortening of action potential duration (APD) of guinea pig papillary muscle; on average, APD<SUB>30</SUB> and APD<SUB>90</SUB> were shortened by 23% and 17% at a stimulation rate of 1 Hz, respectively. Sibutramine suppressed the following currents: I<SUB>hERG</SUB> (IC<SUB>50</SUB>:2.408 ± 0.5117 μg/mL), L-type Ca current (IC<SUB>50</SUB>:2.709 ± 0.4701 μg/mL), and Na current (IC<SUB>50</SUB>:7.718 ± 1.7368 μg/mL). Sibutramine blocked I<SUB>hERG</SUB>, I<SUB>Ca</SUB>, and I<SUB>Na</SUB> in a concentration-dependent manner. In conclusion, sibutramine exerted a shortening effect on APD in guinea pig papillary muscle through its more powerful blocking effects on I<SUB>Ca</SUB> and I<SUB>Na</SUB> rather than I<SUB>hERG</SUB>.</P>

자궁경부암 환자에서 ELISA를 이용한 HPV-16 VLP에 대한 혈청반응

김승조,남궁성은,박종섭,김현수,김찬주,박순희,안희경,황은성,정근택 대한산부인과학회 1996 Obstetrics & Gynecology Science Vol.39 No.1

1993년 1월부터 1994년 12월까지 가톨릭대학교 의과대학 강남성모병원 산부인과 종양학부에서 진단 및 치료를 받았던 환자들에서 치료전 채취한 혈청과 자궁경부 세포를 이용한 실험결과 다음과 같은 결론을 얻을 수 있었다. 1. 자궁경부암 전구병변 환자 20명은 HPV-16에 양성이었고, 나머지 4명은 다른 아형의 DNA HPV가 있거나, 발견되지 않았다. 96명의 자궁경부암 환자에서 69명은 HPV-16 DNA, 9명은 HPV-18 DNA에 양성이었고, 그외 3명은 HPV-16과 HPV-18이 동시에 발견되었다. 또한 7명에서는 다른 아형의 HPV DNA가 있었고, 나머지 8명은 HPV DNA를 발견할 수 없었다. 2. HPV-16 L1, L2 VLP에 대한 ELISA 결과는 정상 대조군의 9%에서 양성이었고, 자궁경부 전구병변의 환자에서는 50%(12/24), 자궁경부암 환자의 62.5%에서 각각 양성으로 나타나 자궁경부의 종양성 변화가 있는 환자에서 유의한 진단적 가치가 있었다. 3. HPV-16 DNA에 양성인 자궁경부암 환자의 68%(47/69)에서 HPV-16 VLP ELISA에 양성이었고, HPV-18 DNA에 양성인 환자의 44%(4/9)에서 HPV-16 VLP ELISA에 양성이었으나 HPV-16/18이외의 다른 HPV 아형에 양성인 환자에서는 14%(1/7)에서 ELISA 양성으로 나타나서 HPV-16또는 HPV-18 DNA 양성인 환자들과 다른 종류 HPV DNA에 양성인 환자들 사이에서 항체 반응 여부는 통계학적인 차이가 있었다. 4. 자궁경부암 환자의 임상기와 조직학적 특성에 따른 HPV-16 VLP에 대한 항체 형성을 관찰해 본 결과 각 임상기와 조직형에 따른 차이를 관찰할 수 없었다. 이상의 결과로 자궁경부암과 전구 병변 환자들의 대다수에서 ELISA 방법으로 HPV-16 VLP에 대한 항체 형성을 관찰할 수 있었다. 또한 HPV-16 VLP ELISA 검사는 HPV-16 DNA 검사의 보조진단법으로 유용하며, HPV와 연관된 자궁경부 종양성 질환의 혈청학적 역학 연구에도 도움을 줄 수 있을 것으로 사료된다. Background: The late structural proteins of HPV-16 have recently been shown to self-assemble into virus-lide particles(VLPs) from baculovirus system when expressed in insect cells. The ability to generate preparative amounts of HPV-16 L1-L2 VLP may have implications for the development of a serologic assay to detect anti-HPV-16 virion immune responses to conformational epitopes and for immunoprophylasis against HPV-16 infection. Methods: We attempted to investigate serologic responses in the sera obtained from Korean women with cervical neoplasia by ELISA using HPV-16 VLPs. PCR method using L1 consensus primers and type-specific oligomer probes was used to determine the presence and type of HPV infection in the study population(normal control; 23 cases, preinvasive lesions; 24 cases, and invasive cervical cancers; 96 cases). Result: 1) The sera of 93%(21/23) of women, who didn`t have HPV-ralated cervical lesions, were negative for HPV-16 VLPs in ELISA assay. The sera of 50%(12/24) of patients with preinvasive cervical lesion and 63%(60/96) of patients with invasive cervical cancer were ELISA positive for HPV-16 VLPs(p$lt;0.01). 2) The positive reactivities for ELISA of HPV-16 VLPs in cervical cancer patients according to clinical characteristics were nor signific antly different. 3) Among women with cervical neoplasia, HPV 16 VLP was positive in 50%(10/20) of HPV-16 positive cases with preinvasive cervical lesion and 72%(50/72) of HPV-16 positive cases with invasive cervical cancer. Conclusion: These results suggest that the considerable number of patients with cervical neoplasia generated an IgG antibody response to the HPV 16 VLPs that can be detected by ELISA. These HPV-16 VLP ELISA may be useful in adjunctive diagnostic asay to HPV-16 DNA test and seroepidemiologic study of HPV-related cervical neopl-asia.