자궁경부 종양에서 p53, Proliferating Cell Nuclear Antigen(PCNA), c-myc 및 Epidermal Growth Factor Receptor(EGFR)의 발현

한구택 ( Ku Taek Han ),이형근 ( Hyung Kun Lee ),유순원 ( Sun Won Yoo ),송영훈 ( Young Hun Song ),최원영 ( Won Young Choi ),유기성 ( Ki Sung Ryu ),나종구 ( Jong Gu Rha ),송승규 ( Seung Kyu Song ),이헌영 ( Hun Young Lee ),김수평 ( S 대한산부인과학회 1997 Obstetrics & Gynecology Science Vol.40 No.12

Expression of four biologic markers known as p53, proliferating cell nuclear antigen(PCNA), c-myc, and epidermal growth factor receptor(EGFR), was studied immunohistochemically in 121 cases of cervical tissues, which included 15 cases of normal cervical tissues, 50 cases of squamous intraepithelial lesions(SIL), and 56 cases of invasive carcinomas. This study was designed to elucidate the roles of these factors in the genesis and progression of cervical neoplasia and to identify their association with clinical parameters such as cell type, tumor size, lymph node involvement, squamous cell carcinoma(SCC) antigen level, and prognosis. In additon, this study evaluates the differences of coexpression rates in normal cervical tissues, SILs, and invasive carcinomas. (1) The intensities of p53, c-myc, and EGFR expression and the rates of PCNA, c-myc, and EGFR expression in cervical SILs and invasive carcinomas were significantly higher than those in normal tissues (p<0.05 and p<0.01). But there were no significant differences between SILs and invasive carcinomas in both of intensities and rates of p53, PCNA, c-myc, and EGFR expression. (2) The intensities and rates of p53, PCNA, and c-myc expression in the invasive carcinomas did not correlate with clinical parameters including cell types, tumor sizes, lymph node involvements, see antigen, and prognosis. But only the intensity and rate of EGFR expression were significantly higher in cases with tumor size larger than 3 cm when it was compared with cases with tumor sizes smaller than 3 cm(p<0.01), in cases with lymph node involvement when it was compared with cases without lymph node involvement(p<0.05), and in cases with SCC antigen levels more than 2.5 ng/ml when it was compared with cases with SCC antigen levels less than 2.5 ng/ml(p<0.05). (3) C-myc immunoreactivity was significantly correlated with EGFR overexpression(p<0.01). (4) The rates of simultaneous expression of four factors were decreased significantly in normal tissues and significantly increased in invasive carcinomas(p<0.01). The coexpression rate of two factors or more was 6.7% in normal tissues, 62.7% in SILs, and 70.0% in invasive carcinomas and the simultaneous expression rate of three factors or more was none in normal tissues, 34.0% in SILs, and 47.5 % in invasive carcinomas. These results show that the immunohistochemical detection of p53, PCNA, c-myc, and EGFR expression will be useful in differentiating the normal tissues and cervical neoplasia. The results of simultaneous immunohistochemical detection of these four factor expression also suggest to contribute a better understanding of the genesis of cervical carcinoma and that coexpression of more factors indicate more aggressiveness in cervical neoplasia, and show that simultaneous detection of these factors can be used in the early detection of cervical neoplasia and to predict malignant transformation of cervical lesions.

항암제를 투여한 세포주의 배양액에서 M30을 이용한 고사의 측정

박상호 ( Sang Ho Park ),곽현성 ( Hyun Sung Kwack ),류기성 ( Ki Sung Ryu ),이영 ( Young Lee ),한구택 ( Ku Taek Han ) 대한산부인과학회 2010 Obstetrics & Gynecology Science Vol.53 No.5

목적: 부인종양에서 실시되는 신보조항암화학요법의 효율성을 증가시키기 위해서는 약제들에 대한 반응을 조기에 예측할 수있는 측정법이 절실히 요구되고 있다. 본 연구에서는 부인종양의 치료에 흔히 사용되는 항암화학요법제의 투여 후에 배양된 세포들과 이들의 배양액내에서 유발된 고사의 정도를 M30 항원을 이용하여 측정함으로써 약제에 대한 반응의 생화학적 지표로서 사용 가능성이 있는지를 알아보았다. 연구 방법: 배양한 HeLa와 OVCAR-3 세포주에 paclitaxel, cisplatin 및 camptothecin의 저농도와 고농도를 각각 투여한 후 수확한 세포들과 배양액내에서 면역형광염색법과 ELISA에 의해 M30 항원을 측정하였다. 결과: 세포질과 배양액내에서 M30 항원의 정도를 각각 측정한 결과는 비투여 대조군으로부터 항암요법제의 투여 농도가 저농도와 고농도로 증가됨에 따라 M30 항원의 수준이 점차적으로 유의하게 증가함을 관찰하였다. HeLa와 OVCAR-3 세포들에서 M30-fluoresein isothiocyanate (FITC)의 면역형광이 양성인 세포들은 대조군에서 각각 4.3%와 4.07%였다. 약제의 농도 (저농도 vs 고농도)에 따라 paclitaxel의 경우에는 18.61%와 18.6% vs 34.87%와 32.63%로 증가되었고, cisplatin의 경우에는 17.87%와 16.83% vs 32.38%와 32%로 증가되었으며, camptothecin의 경우에는 17.87%와 16.83% vs 32.38%와 32%로 점차 유의하게 증가하는 모습이 괄찰되었다 (P<0.001). 또한 HeLa와 OVCAR-3 세포들의 배양액내에서 ELISA에 의해 측정된 M30 항원의 값은 대조군에서 각각 53.03 U/L와 86 U/L이었다. 약제의 농도 (저농도 vs 고농도)에 따라 paclitaxel의 경우에는 101.53 U/L와 114 U/L vs 355.59 U/L와 412.04 U/L이었고, cisplatin의 경우에는 79.84 U/L와 125.44 U/L vs 327.64 U/L와 385.09 U/L이었으며, camptothecin의 경우에는 88.41 U/L와 108.42 U/L vs 295.005 U/L와 263.1 U/L로 증가함을 관찰하였다 (P<0.001). 결론: 본 연구를 통해 HeLa와 OVCAR-3 세포주에서 paclitaxel, cisplatin과 camptothecin을 투여한 후 수확한 세포들과 배양액내에서 고사의 지표로서 측정한 M30 항원이 증가한 결과들은 암세포의 항암요법제에 대한 감수성의 조기 평가에 유용할 것으로 생각되었다. Objective: We investigated a possible use of the induced apoptosis as a biomarker in the cells and their media treated with commonly used anti-cancer agents in gynecologic malignancies. Methods: After treatments with low and high concentrations of paclitaxel, cisplatin, and camptothecin in HeLa and OVCAR-3 cells, the levels of M30 antigen were detected in the cells and their media by immunofluorescence staining and ELISA methods, respectively. Results: The percentages of M30-fluoresein isothiocyanate (FITC) positive cells in HeLa and OVCAR-3 cells treated with paclitaxel, cisplatin, and camptothecin were 4.3% vs 18.1% vs 34.87% and 4.07% vs 18.6% vs 32.63%, 4.3% vs 17.87% vs 32.38% and 4.07% vs 16.83% vs 32%, and 4.3% vs 16.75% vs 31.3% and 4.07% vs 15.18% vs 29.9% in control, low dose, and hight dose groups, respectively (P<0.001). M30 antigen levels (U/L) measured in culture media of HeLa and OVCAR-3 cells treated with paclitaxel, cisplatin, and camptothecin were 53.03 vs 101.53 vs 355.59 and 86 vs 114.41 vs 412.04, 53.03 vs 79.84 vs 327.64 and 86 vs 125.44 vs 385.09, and 53.03 vs 88.41 vs 295.005 and 86 vs 108.42 vs 263.1 in control, low dose, and hight dose groups, respectively (P<0.001). Conclusion: Our results obtained in this preclinical study suggests that measurement of the levels of M30 antigen may help to predict the clinical responses and to select the effective anti-cancer agents in clinical settings, rapidly and quantitatively.

항암제를 투여한 HeLa와 OVCAR-3 세포주에서 Cytokeratin 18-Asp(396) (M30) 항체를 이용한 항암제 감수성의 측정

송민경 ( Min Kyung Song ),박상호 ( Sang Ho Park ),곽현성 ( Hyun Sung Kwack ),류기성 ( Ki Sung Ryu ),한구택 ( Ku Taek Han ) 대한산부인과학회 2010 Obstetrics & Gynecology Science Vol.53 No.1

목적: 항암화학요법제의 투여로 유발되는 세포고사의 과정에서 cytokeratin 18의 분해 산물인 cytokeratin 18-Asp(396) (M30) 단편의 노출된 신항원 결정기와 특이적으로 결합하는 M30 단클론항체를 이용한 세포고사 측정이 자궁경부암과 난소암에서 신보조화학요법의 감수성을 평가하기 위한 표지자로서의 사용가능성을 알아보기 위해 본 실험을 실시하였다. 연구 방법: 1,000 nM의 paclitaxel, 250 μg/mL의 cisplatin과 30 μM의 camptothecin의 투여와 함께 배양한 HeLa와 OVCAR-3 세포주들에서 M30 단클론항체를 이용한 면역형광염색 실시 후, 유세포분석에 의해 세포주기의 변화와 세포고사의 정도를 측정하였으며, 또한 이들의 배양액에서 M30 단클론항체를 이용한 ELISA법으로 세포고사의 정도를 측정했다. 결과: Paclitaxel 투여군에서 HeLa와 OVCAR-3 세포주 모두에서 대조군에 비해 G0G1이 감소되었고, G2M 분기 분획은 증가하였다. Cisplatin 투여군에서 두 세포주에서 G0G1과 S 분획이 대조군과 차이가 없었으며, HeLa 세포주에서는 G2M 분기 분획이 감소되고 OVCAR-3 세포주에서는 감소되는 경향이 관찰되었다. Camptothecin 투여군에서 S와 G2M 분기 분획이 두 세포주 모두에서 대조군과 차이가 없었으며, HeLa 세포주에서는 G0G1 분기 분획이 유의한 증가를 나타냈고, OVCAR-3 세포주에서는 G2M 분기 분획이 감소된 경향을 보였다. G0G1하 첨봉 (Sub-G0G1 peak)에 의해 측정된 세포고사의 정도는 paclitaxel 투여군에서 HeLa와 OVCAR-3 세포주 모두에서 차이가 없었으나, cisplatin 투 여군에서 OVCAR-3 세포주에서 유의하게 증가되었으며, camptothecin 투여군에서는 두 세포주 모두에서 유의하게 증가되었다. M30-FITC 형광도와 배양액 내에서 ELISA법으로 측정된 M30에 의한 세포고사의 정도는 모든 투여군에서 대조군보다 증가되었다. 결론: HeLa와 OVCAR-3 세포주들에서 paclitaxel, cisplatin과 camptothecin을 투여 후 다변수 유세포분석, 면역형광염색 및 ELISA법에 의해 세포주기 변화와 G0G1하 첨봉 및 M30 항원을 측정한 결과 세포고사의 유의한 증가가 확인됐으며, M30 항원의 측정이 좀 더 유의한 반응을 보였다. Objective: The aim of this study was to detect the levels of M30-antigens as a biomarker of apoptosis in cells and their culture media after treatments with anticancer drugs as a preclinical study. Methods: After HeLa and OVCAR-3 cells were treated respectively with paclitaxel, cisplatin, and camptothecin, the harvested cells were stained sequentially with M30 monoclonal antibodies and propidium iodide (PI). Afterwards, they were analyzed using a FACScan flow cytometer and observed under an immunofluorescence microscope for M30-FITC immunofluorescences. Levels of M30 antigens were also detected in their culture media using M30-Apoptosense ELISA kit. Results: The levels of M30-FITC immunofluorescences were elevated in both cell lines after each drug treatments compared with those of control cells. The levels of M30 antigens detected by ELISA in media culturing each cell line treated with each of drugs were elevated compared with those of control cells. Conclusion: This study suggests that M30-antigens representing chemotherapy induced apoptosis may be a useful biomarker for predicting and monitoring the response of neoadjuvant chemotherapy in patients with gynecologic cancers.

대음순에서 발생한 점액성 신경섬유종

곽현성 ( Hyun Sung Kwack ),박호정 ( Ho Jung Park ),임선옥 ( Sun Ok Im ),류기성 ( Ki Sung Ryu ),한구택 ( Ku Taek Han ) 대한산부인과학회 2009 Obstetrics & Gynecology Science Vol.52 No.1

Myxoid neurofibroma is a benign tumor of perineural cell origin, which is demonstrated with a positive immunohistochemical stainig for S-100. The most common locations of the myxoid neurofibroma are face, shoulder, arm, periungual and foot. We experienced an unusual case of myxoid neurofibroma which originated from labia majora area then reported it. To our knowledge, this is the first time that a labia majora location is reported.

MCF-7 세포주에서 Staurosporine 투여에 의한 세포주기 분기 변화와 p53단백 발현

남정(Jung Nam),여경아(Kyung A Yea),이해남(Hae Nam Lee),조현희(Hyun Hee Jo),류기성(Ki Sung Ryu),유영옥(Young Oak Lew),나종구(Jong Gu Rha),한구택(Ku Taek Han) 대한산부인과학회 2001 Obstetrics & Gynecology Science Vol.44 No.3

N/A Objective : We investigated the effects on the cell cycle and p53 expression by the treatment of various concentrations of staurosporine to elucidate the molecular mechanism of staurosporine induced cell cycle arrest in MCF-7 cell line. Methods : Various concentrations of staurosporine were treated in MCF-7 cells cultured with RPMI 1640 media. The harvested cells were fixed and permeabilized with 1% paraformaldehyde and absolute methanol. Then the cells were stained indirectly with anti-p53 primary antibody and FITC conjugated goat anti-mouse(GAM)-IgG secondary antibody. Sequentially DNA were stained with 0.1% RNase and PI solution. These stained cells were analyzed by the standard FACScan flow cytometer. The obtained results were analyzed further with WinList 3.0, and ModiFit LT software program. Results : MCF-7 cells were arrested mostly in G1 phase of cell cycle at 5-10 nM of staurosporine, however, the cells were arrested in G2 phase at 20-100 nM of staurosporine. The expressions of p53 protein were higher in the MCF-7 cells treated with both concentrations of 10 nM and 100 nM of staurosporine compaired with the control cells. This suggests that the p53 may be involved in the mechanism of G1 and G2M arrest of cell cycle in MCF-7 cell. Conclusion : The points of arrest in cell cycle differred depending on the concentrations of staurosporine and these cell cycle arrests at G0G1 and G2M pahse were related with p53 protein expression. It suggested that these results could be extended to study for staurosporine to be usefull as a potential anti-tumor agent.

난소 종양에서 Mullerian Inhibiting Substance와 그 수용체의 발현

서경아 ( Kyoung A Seo ),류기성 ( Ki Sung Ryu ),이정원 ( Chung Won Lee ),최미나 ( Mi Na Choi ),차정호 ( Jung Ho Cha ),김장흡 ( Jang Heub Kim ),한구택 ( Ku Taek Han ) 대한산부인과학회 2005 Obstetrics & Gynecology Science Vol.48 No.2

목적: 임신 7주경 태아 고환의 미숙 Sertoli 세포에서 생산되어 Muller관을 퇴행시키는 Mullerian inhibiting substance (MIS)는 가임기 여성의 생식 생리 기전을 조절하고 일부 난소암 세포주의 성장을 억제하는 작용이 있음이 밝혀졌다. 그러나 대부분의 연구가 설치류를 대상으로 하였으며, 아직도 인간의 정상 생식 생리 기전이나 난소 종양의 발생 과정에 대한 연구는 미흡한 상태이다. 이에 본 연구자들은 MIS와 MIS typ Mullerian inhibiting substance (MIS) is a glycoprotein hormone produced by fetal Sertoli cells that causes regression of the Mullerian ducts in males during sexual differentiation. Cell lines derived from human ovarian epithelium and rodent Leydig cell tu

절제수술로 치료한 자궁경부 상피내암과 미세침윤암에서 세포진과 조직진의 비교평가

윤수철 ( Soo Cheol Yoon ),공기환 ( Gi Hwan Gong ),정선안 ( Sun An Jung ),최영석 ( Young Seuk Choi ),유기성 ( Ki Sung Ryu ),한구택 ( Ku Taek Han ),나종구 ( Jong Gu Rha ),이헌영 ( Hun Young Lee ),김수평 ( Soo Pyung Kim ) 대한산부인과학회 1997 Obstetrics & Gynecology Science Vol.40 No.12

We evaluated 221 patients who were preoperatively diognosed as carcinoma in situ(CIS) and microinvasive carcinoma(MIC) from Jan. 1988 to Dec. 1995 at the Department of Obstetrics and Gynecology, St. Mary`s hospital, Catholic University Medical College. The data gained from 221 patients with sucessful tollow-up were thoroughly reviewed to determine the proper diagnosis and treatment model. They were treated by conization, total abdominal hysterectomy or type II hysterectomy. The final diagnoses of 127 patients who got one cytologic diagnosis at local clinic or our hospital were 80 CIS, 45 MIC and 2 mvasive carcinomas. In cytology, 47 case(58.8 %) of 80 CIS showed abnormal findings as class III(30 %), IV(22.5 %) and V(6.3 %). In 45 MIC, 33(73.3 %) cases showed abnormal finding as class III(35.6 %), IV(28.9 %) and V(8.9 %). 94 patients who got cytology twice at the local clinic or our department were finally diagnosed as 72 CIS and 22 MIC. By repeating cytology, we could reduce false positive rate, 12.5 % in CIS and 4.5 % in MIC, however repeat cytology had little diagnostic value. In comparison of colposcopy guided biopsy(CGB) to non-colposcopic blind biopsy, CGB was more predictive. In CIS, the accurate diaganosis rates were 77.6 %(52/67) in blind biopsy and 88.8 %(95/124) in CGB. In MIC, the accuracies were 71.8%(28/39) in blind biopsy and 100 %(33/33) in CGB. So colposcopic evaluation with optional conization was essential for pooper diagnosis and treatment. There were unfavorable histologic findings: vaginal extension(n=4; 1.8 %), lymphovascular space involvement(n=4; 1.8 %), perineural invasion(n=1; 0.5 %) and occult invasive cancer(n=2; 1.8 %). So conservative treatment and local destructive therapy were potentially dangerous in CIS and MIC and should be performed after precise evaluation in selected cases. For proper diagnosis and treatment, cytologic evaluation had limited clinical values and CGB with optional conization was needed.

DES (Diethyl stilbestrol)에 노출된 경험이 없는 27세 여성에서 발생한 자궁경부 투명세포암 및 근치적자궁경부절제술 (Radical Trachelectomy)

곽현성 ( Hyun Sung Kwack ),문영주 ( Young Joo Mun ),하재은 ( Jae Eun Ha ),황선정 ( Sun Jung Hwang ),김수미 ( Su Mi Kim ),한구택 ( Ku Taek Han ),류기성 ( Ki Sung Ryu ) 대한산부인과학회 2009 Obstetrics & Gynecology Science Vol.52 No.9

Clear cell adenocarcinoma (CCAC) is a rare cancer that comprises less than 9% of the cervical adenocarcinoma cases. We experienced a case of fertility-sparing radical abdominal trachelectomy for cervical clear cell adenocarcinoma (CCAC). Thus, reported it. A 27 year old female was diagnosed with clinical stage Ib cervical CCAC. She had no history of maternal exposure to diethylstilbestrol and had negative PAP cytology and HPV tests. She was treated with neoadjuvant chemotherapy followed by radical abdominal trachelectomy. After 2 cycles of postoperative adjuvant chemotherapy, the lesion disappeared completely in an imaging study, and potential fertility was preserved. Radical abdominal trachelectomy with chemotherapy may be a valuable approach for treating stage Ib cervical CCAC in women that wish to preserve potential fertility.

난소암에서 자가 골수이식과 말초혈액 지지에 의한 고용량 항암 화학요법

한구택 대한조혈모세포이식학회 1996 대한조혈모세포이식학회지 Vol.1 No.1

고형 종양 특허 난소암에서 고용량의 항암 화학요법의 시도는 전 세계적으로도 매우 적으나, 진행성 난소암에서 사용되어온 표준 항암 화학요법제의 약제반응과 장기간의 무병 생존율은 20% 정도이지만 내성을 일으켰거나 진행된 난소암 환자에서 현재 까지 시도된 자가 골수 이식 지지나 말초 혈액 선조세포 지지에 의한 고용량의 항암 화학요법의 결과들은 약 70-80%의 매우 높은 반응률을 보였다. 이러한 결과들은 고용량의 항암제와 골수 또는 말초 혈액 원조 세포 지지 요법에 대한 약속된 결과를 암시하는 것이다. 그러나 비록 종양의 용적의 감소는 나타났지만 9개월 정도로 보고 된 짧은 평균 반응 기간은 실제적으로는 잔존 종양이 남아있음을 암시한다. 따라서 질환의 경과 중에서 종양의 내성이 덜 나타나는 더 조기에 고용량의 화학요법을 적용한다면 좀 더 지속되는 항종양 효과를 얻을 수 있을 것이다. 그러나 자기골수이식 지지와 말초혈액 선조세포 지지에 의한 고요량 항암 화학요법은 이환율과 사망율의 관점에서 많은 비용을 필요로 하고 여러 전문분야(multidisciplinary effort)의 노력 이 필요하므로 일률적으로 치명적인 질환에서 평균 무재발 생존률 또는 전체 생존률의 연장의 시위보다는 더 나은 목표를 설정해야 하며 백혈병이나 림프종에서와 같이 많은 환자들에서 장기간 지속되는 무병생존(완처)의 성취가 궁국적인 목표이어야 할 것으로 생각되며 본 산부인과학 교실에서도 내과학 교실과 골수이식 센터의 도움으로 고요량 하암도법의 도입이 하루 빨리 이루어지기를 기대한다.

부인암 환자의 항암화학요법 후 혈액학적 관찰

이승희,김재훈,이헌영,손우익,김휘,송경근,김현홍,한구택 대한부인종양 콜포스코피학회 1993 Journal of Gynecologic Oncology Vol.4 No.3

A retrogpective review of hematologic monitoring involving aggressive chemotherapy was careiyd out ta assess whether there is a predictable relationship between the white blood cell count end the platelet count as a reflection of bone marrow toxicity and when maximum myeloauppression occur during a treatment program. This data revealed that the white blood cell and granulocyte levels are closely related and that myeloeuppression can occur during any course of CAP(cyclophosphamide, adriamycin, and cisplatin), VBP(vinblastine, bleomycin, and cisplatin) chemotherspy in gynecological cancer. Thus, for these treatment regimens in gynecoldgical malignancies, the white blood cell and granulocyte count is sufficient for monitoring toxicity and adjusting future courses of chemotherapy. There are no bone marrow depressions by the treatment regimens for the gestational trophoblastic disease.