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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
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.
인간 난소에서 생리 주기에 따른 Mullerian Inhibiting Substance와 그 수용체의 발현
김장흡 ( Jang Heub Kim ),정서호 ( Seo Ho Chung ),최은주 ( Eun Joo Choi ),황성진 ( Hwang Seong Jin ),조현희 ( Hyun Hee Jo ),김미란 ( Mee Ran Kim ),김은중 ( Eun Jung Kim ),김진홍 ( Jin Hong Kim ),류기성 ( Ki Sung Ryu ) 대한산부인과학회 2004 Obstetrics & Gynecology Science Vol.47 No.9
목적 : Mullerian inhibiting substance (MIS)의 주 생산 장기인 가임기 여성 난소에서 MIS와 MIS가 결합하는 MIS type II 수용체 (MISR II)의 정확한 발현위치와 월경에 따른 변화를 알아보고 생식생리에 미치는 역할을 밝히고자 연구를 시행하였다. 연구 방법 : 월경주기와 난소기능이 정상이면서 난소종양이 없는 부인과 환자의 난소조직을 월경주기별 (난포기, 배란기, 황체기)로 구분하여 각각 7예씩 총 21예를 대상 In this study, in order to further understanding of function of Mullerian inhibiting substance (MIS) and the ontogeny of the production profile of biologically active MIS and MIS type II receptor (MISR II), the patterns of their localization according to
( Jang Heub Kim ),( David T Maclaughlin ),( Patricia K Donahoe ) 대한산부인과학회 2014 Obstetrics & Gynecology Science Vol.57 No.5
Mullerian inhibiting substance (MIS), also called anti-Mullerian hormone (AMH), is a member of the transforming growth factor-β super-family of growth and differentiation response modifiers. It is produced in immature Sertoli cells in male embryos and binds to MIS/AMH receptors in primordial Mullerian ducts to cause regression of female reproductive structures that are the precursors to the fallopian tubes, the surface epithelium of the ovaries, the uterus, the cervix, and the upper third of the vagina. Because most gynecologic tumors originate from Mullerian ductderived tissues, and since MIS/AMH causes regression of the Mullerian duct in male embryos, it is expected to inhibit the growth of gynecologic tumors. Purified recombinant human MIS/AMH causes growth inhibition of epithelial ovarian cancer cells and cell lines in vitro and in vitro via MIS receptor-mediated mechanism. Furthermore, several lines of evidence suggest that MIS/AMH inhibits proliferation in tissues and cell lines of other MIS/AMH receptor-expressing gynecologic tumors such as cervical, endometrial, breast, and in endometriosis as well. These findings indicate that bioactive MIS/AMH recombinant protein should be tested in patients against tumors expressing the MIS/AMH receptor complex, perhaps beginning with ovarian cancer because it has the worst prognosis. The molecular tools to identify MIS/AMH receptor expressing ovarian and other cancers are in place, thus, it is possible to select patients for treatment. An MIS/AMH ELISA exists to follow administered doses of MIS/AMH, as well. Clinical trials await the production of sufficient supplies of qualified recombinant human MIS/AMH for this purpose.
난포발달에 따른 Mullerian Inhibiting Substance Type 2 수용체의 발현
김장흡 ( Jang Heub Kim ),정서호 ( Seo Ho Chung ),황성진 ( Seong Jin Hwang ),조현희 ( Hyun Hee Jo ),김미란 ( Mee Ran Kim ),권동진 ( Dong Jin Kwon ),유영옥 ( Young Ok Lew ),김진홍 ( Jin Hong Kim ),이진우 ( Jin Woo Lee ) 대한산부인과학회 2004 Obstetrics & Gynecology Science Vol.47 No.11
점막하 근종의 진단에 있어서의 초음파 자궁조영술의 유용성에 대한 자궁경 검사를 이용한 고찰
김용식(Yong Sik Kim),위승길(Seung Kil We),오성택(Sung Tack Oh) 대한산부인과학회 2001 Obstetrics & Gynecology Science Vol.44 No.10
N/A Objective : To evaluate the efficacy of sonohysterography on exact submucosal myoma. Methods : Transvaginal sonohysterography was performed on 45 patients who diagnosed as submucosal myoma and 29 patients who diagnosed as endometrial polyp by classic abdominal or vaginal sonogram, and then they received the diagnostic hysteroscopy at all. Results : In 24 of 45 patients on sonohysterogram, submucosal myoma was diagnosed that was protruded to uterine cavity more than 2/3 of mass. Nine patients had intramural myoma and 12 patients had myoma that protruded only lesser than 1/3 of mass on sonohysterogram. These 21 patients revealed the same finding on hysteroscopy. On hysteroscopy, submucosal myomas were in 20 of 24 patients and polyps were in 3 patients and synechia was in one patients. In 8 of 29 patients on sonohysterogram, endometrial hyperplasia was found and confirmed by hysteroscopy. In 16 of 21 patients whose finding was polyp on sonohsyterogram, polyp was confirmed by hysteroscopy. However hysteroscopic findings were myomas in 3 of 21 patients and synechia in one patients. Therefore in 29 of 74 patients (39.2%), further hysteroscopy was not needed by sonohysterogram. In 65 of 74 patients (87.8%), sonohysterographic findings were same as hysteroscopic findings. Conclusion : Sonohysterogram for diagnosis of submucosal myoma and polyp is essential procedure in order to avoid unnecessary hysteroscopy. However differentiation between polyp and submucosal myoma has still some difficulty.
Juhun Lee,Jong Mi Kim,Gun Oh Chong,Dae Gy Hong,Yoon Hee Lee 영남대학교 의과대학 2023 Yeungnam University Journal of Medicine Vol.40 No.-
Background: Over the last two decades, serum levels of anti-Müllerian hormone (AMH) have been shown to be reliable markers of ovarian reserve. This study aimed to compare baseline serum AMH levels and well-controlled clinical factors between patients with unilateral and bilateral ovarian endometriomas during the menstrual phase. Methods: We conducted a retrospective study. We enrolled 136 patients aged 18 to 36 years who were diagnosed with unilateral or bilateral ovarian endometriomas. Serum AMH levels of all patients and their latest two to three menstrual cycles were measured before surgery for ovarian endometriomas. The latest menstrual cycle length ranged from 26 to 30 days. Patients with irregular menstruation, a recent medication history of hormonal drugs other than oral contraceptive pills, a previous history of ovarian surgery, or any medical history influencing ovarian function were excluded. Results: Of the 136 patients, 76 (55.9%) had unilateral ovarian endometriomas and 60 (44.1%) had bilateral ovarian endometriomas. Serum AMH levels were not significantly different between the two groups in the follicular phase, luteal phase, or at any random time point. Conclusion: Serum AMH levels were not significantly different between unilateral and bilateral ovarian endometriomas in the follicular and luteal phases, or at any random time during the menstrual cycle when various confounding factors were excluded.