Development of immunoassay using novel human papapillomavirus 16 E7 antibody

류기성 ( Ki Sung Ryu ),( Keun Ho Lee ),( Hee Jung Lee ),( Chan Joo Kim ),( Heun Soo Kang ),( Jong Sup Park ) 대한산부인과학회 2004 대한산부인과학회 학술대회 Vol.90 No.-

난소암 세포주에서 Mullerian Inhibiting Substance의 증식 억제 효과

류기성 ( Ki Sung Ryu ),서미영 ( Mi Young Seo ),조윤성 ( Yun Sung Jo ),김미란 ( Mee Ran Kim ),김진우 ( Jin Woo Kim ),한구택 ( Goo Taik Han ),이준모 ( Joon Mo Lee ),김장흡 ( Jang Heub Kim ) 대한산부인과학회 2006 Obstetrics & Gynecology Science Vol.49 No.11

목적: 임신 7주경 태아 고환의 미숙 Sertoli 세포에서 생산되어 Muller관을 퇴행시키는 Mullerian inhibiting substance (MIS)는 성 발달과 가임기 여성의 생식생리 기전을 조절하는 이외에 Muller관으로부터 발생하는 일부 종양과 세포주의 성장을 in vivo와 in vitro에서 억제한다는 사실이 밝혀졌다. 이에 본 연구자들은 난소상피암 세포주들에서 MIS type II 수용체 (MISR II)의 발현을 확인하고, MIS를 투여하여 MIS의 세포 증식 억제효과와 그 기전을 밝힘으로서 종양치료제로서 MIS의 임상적 사용에 대한 가능성을 알아보고자 하였다. 연구 방법: 난소암 세포주인 SKOV-3, OVCAR-3 및 OVCAR-8에서 면역조직화학염색법으로 MISR II의 발현여부를 확인하였다. 난소암 세포주의 생존도는 고순도 MIS를 투여하여 24시간과 48시간동안 배양한 후 MTT assay로 측정하였다. MIS에 의한 세포주기 변화는 DNA 염색 후 flow cytometer로 분석하였으며 세포자멸사는 annexin-V-FITC 염색방법을 이용하여 flow cytometer로 평가하였다. MIS에 의한 난소암 세포주의 성장억제 감수성 차이와 세포자멸사 기전을 알아보기 위해 western blot 분석법을 이용하였다. 결과: 모든 난소암 세포주에서 MISR II 발현이 관찰되었지만, 발현정도는 OVCAR-8에서 제일 강했다. 세포 생존도는 OVCAR-8만이 MIS의 처치 용량과 시간에 따라 감소하였고, OVCAR-3은 고농도 MIS로 48시간 투여한 경우에만 의의 있게 감소하였으나, SKOV-3은 반응하지 않았다. Flow cytometer로 조사한 세포주기 변화는 OVCAR-8에서 10 μg/mL MIS로 24시간 처치한 경우에 G1세포주기 정지 소견을 보였으며, 48시간 후에는 세포자멸사를 의미하는 sub G0G1분기 분획이 7.02%로 나타나, MIS는 세포주기의 변화를 먼저 일으킨 후 세포자멸사를 유발하였다. OVCAR-3에서는 48시간 후 sub G0G1분기가 3.32%였으나 SKOV-3에서는 변화가 없었다. Annexin-V-FITC 염색방법으로, OVCAR-8과 OVCAR-3에서 48시간 MIS 처치 후에 각각 8.05%와 3.67%의 세포자멸사가 관찰되었다. Western blot 분석으로, OVCAR-8에서 MIS에 의한 세포 증식 억제에 이은 세포자멸사는 p16단백 증가와 연관이 있다고 판단된다. 또한 MIS는 OVCAR-8에서 MISR II와 결합한 후 pRb 비의존적인 세포내 경로를 통해 간접적으로 E2F1 활성 증가로 세포자멸사를 유도할 가능성이 있다. 결론: 본 연구에서 MIS가 난소암 세포주에서 증식 억제와 세포자멸사를 일으키는 기전을 완전히 규명하지는 못하였지만, MIS가 MIS 수용체를 발현하는 난소암에 대하여 in vitro에서 효과적인 항종양능을 나타낸다는 사실을 확인할 수 있었다. 향후 더 많은 연구로 MIS가 MIS 수용체를 발현하는 종양의 생물학적 조절제 혹은 치료제로 개발될 것이라고 예상된다. Objective: In order to explore Mullerian inhibiting substance (MIS) effects on the ovarian neoplasia, the expression and localization of the MIS type II receptor (MISR II), the growth inhibitory effects of MIS, and the underlying molecular mechanisms were investigated in the ovarian cancer cell lines. Methods: Expression of MISR Ⅱ were studied in SKOV-3, OVCAR-3, and OVCAR-8 cell lines by immunohistochemical staining. The antiproliferative effects of MIS in these cell lines were investigated by methylthiazoletetrazolium (MTT) assay, fluorescence-activated cell sorting (FACS) analysis, annexin-V-FITC binding, and western blot analysis. Results: All cell lines showed strong specific staining for MISR II, although staining in OVCAR-8 cells was more intense than that in SKOV-3 and OVCAR-3. Treatment of OVCAR-8 cells with MIS led to a dose- and time-dependent inhibition of cell growth and survival was determined use by MTT assay. But OVCAR-3 cells exhibited growth inhibition at higher doses after 48 hours of treatment and SKOV-3 cells did not demonstrate response. Using FACS analysis, exposure of OVCAR-8 cells to MIS (71 nM) resulted in G1 arrest after 24 hours of treatment. This pattern was changed by time-dependent increase in the percentage of cells with a sub G0G1 DNA content, suggesting apoptosis, after 48 hours of treatment. These results suggested that cell death be preceded by cell cycle arrest. Time-related induction of apoptosis was also observed in this cell line as measured by annexin-V-FITC binding. In OVCAR-8 cells, the growth inhibitory effects of MIS were mediated through specific induction of CDKI p16 protein expression and via regulation of E2F1 in the absence of detectable levels of pRb. We estimated that OVCAR-3 cells were affected by MIS through p16-independent, alternative mechanistic pathways, since the growth inhibitory effects of MIS were minimal. SKOV-3 cells did not express p16 protein. Conclusion: We have demonstrated that ovarian cancer cells express the MISR II. Epithelial ovarian cancer cells respond to MIS by growth inhibition. Although the precise mechanisms of MIS mediated inhibition of ovarian cancer cell growth have not been fully defined, these data suggest that MIS has activity against ovarian cancers in vitro and may also be an effective targeted therapy for ovarian cancer.

항사구체 기저막 항체 및 p - ANCA 양성인 급속 진행성 사구체신염 1 예

류정일(Jung Il Ryu),조규현(Kyu Hyun Cho),허성욱(Seong Wook Heo),권현희(Hyun Hee Kwon),조창호(Chang Ho Cho),안기성(Ki Sung Ahn),이인희(In Hee Lee) 대한신장학회 2002 Kidney Research and Clinical Practice Vol.21 No.4

Rapidly progressive glomerulonephritis(RPGN) is clinical syndrome characterized by rapid loss of renal function within several weeks to months, with histologic finding of extensive crescent formation. We report a case of RPGN associated with anti-glomerular basement membrane antibody(anti-GBM Ab) and perinuclear-antineutrophilic cytoplasmic antibody(p-ANCA), which rapidly progressed to chronic renal failure. A 44-year-old male was referred to our hospital for evaluation of pitting edema and proteinuria. Both anti-GBM Ab and p-ANCA were detected in serum. Percutaneous renal biopsy showed many crescents with some fibrinoid materials and heavy deposits of IgG. He was treated with pulse methylprednisolone, followed by oral corticosteroid and cyclophosphamide. In spite of immunosuppressive therapy, his renal function deteriorated rapidly and uremic symptoms including pulmonary edema were aggravated. He was started on hemodialysis and he has received regular hemodialysis without recovery of renal function. Further studies will be needed to determine the clinical significance of combined anti-GBM Ab and ANCA.

기존 원발성 막성 신병증에 합병된 반월상 사구체신염 1 예

임상혁(Sang Hyuk Lim),권현희(Hyun Hee Kwon),박경찬(Kyoung Chan Park),류정일(Jung Il Ryu),유성수(Sung Soo You),박민식(Min Sik Park),조창호(Chang Ho Cho),김용진(Yong Jin Kim),안기성(Ki Sung Ahn),이인희(In Hee Lee) 대한신장학회 2002 Kidney Research and Clinical Practice Vol.21 No.5

Membranous nephropathy (MGN) is a common cause for nephrotic syndrome in adults. Renal failure usually develops gradually in patients with MGN and rapid deterioration of renal function is a rare complication. Moreover, the development of crescentric glomerulonephritis (CGN) as a cause of acute renal failure (ARF) in patients with preexisting MGN is very rare. We report 20-year-old male patient with MGN who presented with ARF due to superimposed CGN. He had been diagnosed as idiopathic MGN and his renal function was maintained within normal range. After 13 months of conservative treatment, ARF was developed and renal biopsy was performed to differentiate the possible causes of ARF. Renal biopsy revealed diffuse crescents formation on preexisting MGN. Anti-GBM antibody and ANCA were not detected. This patient was treated with combination therapy including methylprednisolone pulse therapy followed by high dose steroid and cyclophosphamide. His renal function was improved significantly 2 weeks after treatment. In conclusion, when unexplained ARF in patient with MGN develops, prompt investigation for superimposed conditions including CGN is necessary and repeat renal biopsy should be considered.

결절성 경화증이 동반된 태아 심장 횡문근종

길기철 ( Ki Cheol Kil ),남선영 ( Sun Young Nam ),김수미 ( Su Mi Kim ),하재은 ( Jae Eun Ha ),류기성 ( Ki Sung Ryu ),이영 ( Young Lee ) 대한주산의학회 2009 Perinatology Vol.20 No.4

Fetal cardiac tumors are a rare condition and are histologically benign. Cardiac rabdomyoma is the most common cardiac tumor and may be associated with tuberous sclerosis. Clinical follow-up including monitoring of fetal well-being is warranted as long as the fetus remains asymptomatic and there is no evidence for hydrops fetalis. If obstruction to blood flow and/or early fetal compromise is noted, then the decision of whether to deliver early must be made. We report a case of fetal cardiac tumor diagnosed by antenatal ultrasonography and diagnosed as tuberous sclerosis with brain MRI scanning after birth.

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.

펜타센 TFT의 Flexible OLED 구동

최기범(Ki Beom Choe),류기성(Gi Seong Ryu),변현숙(Hyun Sook Byun),허영헌(Y. X. X),황성범(Sung Beom Hwang),송정근(Chun Kun Song) 제어로봇시스템학회 2004 제어로봇시스템학회 합동학술대회 논문집 Vol.2004 No.12

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.

임신중 골반 기저를 뚫고 질벽으로 탈출한 장막하근종

조윤성 ( Yun Sung Jo ),권지영 ( Ji Young Kwon ),임희순 ( Hee Sun Lim ),문영주 ( Young Joo Mun ),이상형 ( Sang Hyoung Lee ),제동성 ( Dong Sung Jae ),한구택 ( Gu Taek Han ),류기성 ( Ki Sung Ryu ) 대한산부인과학회 2006 Obstetrics & Gynecology Science Vol.49 No.12

Myoma is the most common tumor in gynecologic field. As ultrasonography because popular in antenatal care, the more cases of myoma and those adverse effects during pregnancy are more frequently detected. The management of myoma during pregnancy is conservative, but in rare circumstances, surgical intervention including myomectomy may be required. We have experienced a case of protruded subserosal myoma with the uterine cervix in midtrimester of pregnancy. The patient was managed surgically by transvaginal myomectomy and had successfully maintained pregnancy. We report a case of protruded subserosal myoma through pelvic floor in pregnancy with brief review of literatures.

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.