조기진통 융모막에서 cDNA Microarray를 이용한 유전자 발현 분석

고현선 ( Hyun Sun Ko ),김연희 ( Yeun Hee Kim ),길기철 ( Ki Cheal Kil ),양동은 ( Dong Eun Yang ),박인양 ( In Yang Park ),김사진 ( Sa Jin Kim ),박태철 ( Tai Churl Park ),신종철 ( Jong Chul Shin ) 대한주산의학회 2006 大韓周産醫學會雜誌 Vol.17 No.2

목적 : 조기진통으로 인한 조기분만과 진통을 겪은 만삭분만의 융모막에서 발현되는 gene expression profile을 구별하고자 하였다. 방법 : DNA chip을 이용 조기진통으로 조기분만 5예와 진통을 겪은 만삭분만 5예의 융모막에서 분만직후 각각 RNA를 추출하여 cDNA microarray 실험을 시행하였다. 결과 : 조기 분만시 융모막에서 증가된 발현양상을 보인 유전자는 총 104개 였으며, 이중 1.5배 이상 증가된 것은 총 18개로 cell adhesion, cell cycle regulation, development, transport, morphogenesis, muscle contraction, signal transduction, 및 transcription 등에 관련된 유전자였다. 조기 분만시 융모막에서 유의하게 감소된 발현 양상을 보인 총 102개였고, 이중 1.5배 이상 감소발현을 나타낸 유전자는 총 15개로 cell differentiation, development, metabolism, morphogenesis, RNA processing, signal transduction, transcription과 transport 등에 관련된 유전자였다. 결론 : 본 연구는 cDNA microarray를 이용하면 조기진통에 의한 조기분만의 원인이나 기전을 분자 유전학적 측면에서 더 많이 이해할 수 있을 것이라는 사실을 보여준다. Objective : We used a genome-wide approach to identify differentially expressed genes in patients with preterm delivery caused by preterm labor to improve the understanding of underlying molecular mechanism. Methods : RNA was isolated from the chorions of patients with preterm delivery caused by preterm labor with intact membranes and term vaginal delivery. cDNA microarray experiments were used to identify differentially expressed genes, and reverse transcriptase-polymerase chain reaction was used in follow-up experiment. Results : cDNA microarray experiments identified significantly increased expression of 104 genes and decreased expression of 102 genes in the preterm cases. 18 genes showed more than 1.5-fold increased expressions in the chorions of patients with preterm delivery by preterm labor than those with term vaginal delivery. In preterm delivery, up-regulated genes were associated with cell adhesion, cell cycle regulation, development, transport, morphogenesis, muscle contraction, signal transduction, and transcription. 15 genes showed more than 1.5-fold decreased expressions in chorions of patients delivered preterm by preterm labor than delivered term by labor. In preterm delivery, down-regulated genes were associated with cell differentiation, development, metabolism, morphogenesis, RNA processing, signal transduction, transcription, and transport. Conclusion : This study suggests cDNA micro array technique might provide insights into the molecular basis of preterm delivery caused by preterm labor.

증례: 자기공명영상에 의해 진단된 자궁벽내 임신의 내과적 치료 2예

고현선 ( Hyun Sun Ko ),이영 ( Young Lee ),이희중 ( Hee Joong Lee ),박인양 ( In Yang Park ),정대영 ( Dae Young Chung ),김수평 ( Soo Pyung Kim ),박태철 ( Tai Churl Park ),신종철 ( Jong Chul Shin ) 대한주산의학회 2005 Perinatology Vol.16 No.3

자궁벽내 임신은 자궁외 임신의 1% 미만에서 발생하는 드문 질환으로, 과거에는 수술적 치료가 이루어 졌으나, 최근 자궁에 대한 보존적 치료가 시도되고 있다. 저자들은 임신낭이 파열된 이후 초음파로 진단이 불명확한 경우 자기공명영상을 이용하여 자궁벽내 임신을 진단하고, methotrexate를 통한 내과적 치료를 통해 성공적으로 치료된 자궁벽내 임신 2예를 경험하였기에 문헌고찰과 함께 보고하는 바이다. Intramural pregnancy is one of the rare forms of ectopic pregnancy and the optimal treatment is unknown. We describe two cases of intramural pregnancy treated with systemic methotrexate, which were diagnosed by pelvic MRI. Medical treatment with methotrexate may be a conservative treatment alternative for intramural pregnancies.

조기 분만 태반조직 및 태반 막에서 GeneFishing(TM) DEG PCR 기법을 이용한 유전자 발현 검색

고현선 ( Hyun Sun Ko ),김연희 ( Yeun Hee Kim ),길기철 ( Ki Cheol Kil ),정주희 ( Ju Hee Jeong ),양동은 ( Dong Eun Yang ),정대영 ( Dae Young Chung ),박태철 ( Tai Churl Park ),김수평 ( Soo Pyung Kim ),신종철 ( Jong Chul Shin ) 대한주산의학회 2005 Perinatology Vol.16 No.4

목적: 조기 진통으로 인한 조기분만시 태반 조직 및 태반막에서 GeneFishing(TM) DEG kit의 PCR 기법을 이용한 유전자 발현 양상을 조사하였다. 방법: 39주 이상의 만삭 분만과 32주 미만의 조기 분만시 태반 조직, 양막, 융모막에서 total RNA를 추출하여 GeneFishing(TM) DEG kit를 이용한 PCR을 시행하였다. 발현차이를 보이는 DNA를 정제하여, cloning하 후 ABI HITACHI 3100 genetic analyzer (Perkin-Elmer ABI, Foster City, Calif. USA)로 염기서열을 분석하였다. 분석된 염기서열은 NCBI (<http://www.ncbi.nlm.nih.gov/blast/>)의 blast search를 이용하여 검색하여 유전자를 확인하였다. 결과: 20개의 annealing control primers (ACPs)를 이용하여 발현차이를 보이는 13개의 band를 확인하였고, 그중 조기 분만에서 발현이 증가된 것은 7개, 발현이 감소된 것은 6개였다. BLAST search를 이용하여 확인한 결과 이들 중 각각의 유전자를 대표하는 DNA 절편인 unigene 정보가 있는 유전자는 11개였다. 정보가 있는 유전자 중 조기분만시 발현 증가를 보인 것은 insulin-like growth factor II associated protein, vigilin, acyl-Coenzyme A dehydrogenase, tissue inhibitor of metalloproteinase 1 (TIMP1), ribosomal protein S26 (RPS26), follistatin-like 1 (FSTL1) 이었고, 발현 감소를 보인 것은 두개의 mitochondrial DNAs, ribosomal protein S28 (RPS28), transglutaminase 2 (TGM2), heparin sulfate proteoglycan (HSPG, perlecan) 이었다. 결론: Annealing control primer를 이용한 GeneFishing(TM) DEG kit로 조기분만과 관련된 유전자들을 확인하여 조산의 발생기전을 분자유전학적 측면에서 연구하였으며, 이들 유전자의 기능적 특성에 대한 연구가 필요하다고 생각된다. Objective : The aim of this study was to investigate the gene expression profiles using GeneFishing(TM) kit in human placentae and their membranes delivered at preterm caused by preterm labor. Methods : Specimens were obtained from placenta, chorion, and amnion delivered at preterm and term, respectively. Total RNAs were isolated from each specimen. Thereafter, the profiles of expression genes between preterm and term specimens were compared using a new and accurate reverse transcription-polymerase chain reaction (RT-PCR) that involves annealing control primers (ACPs) to identify the genes expressed differentially and screened by basic local alignment search tool (BLAST) search. Results : Using 20 ACPs, 13 differentially expressed genes (DEGs) were identified and sequenced. 7 of them were expressed up-regulation, while 6 were expressed down-regulation in preterm deliveries. A BLAST searches revealed that 11 were known genes and 2 were unknown genes. Among known genes, up-regulated genes were insulin-like growth factor II associated protein, vigilin, acyl-Coenzyme A dehydrogenase, tissue inhibitor of metalloproteinase 1 (TIMP1), ribosomal protein S26 (RPS26), follistatin-like 1 (FSTL1) and down-regulated genes were two mitochondrial DNAs, ribosomal protein S28 (RPS28), transglutaminase 2 (TGM2), heparin sulfate proteoglycan (HSPG, perlecan). Conclusion : This study shows that the ACP system is a good method for the identification of preterm-related genes. Furthermore, this study suggests that further analysis of the differentially expressed genes in preterm we have identified should provide insights into the molecular basis of preterm delivery caused by preterm labor.

TC-1 세포주를 이용한 동물모델에서 As4O6에 의한 혈관폐쇄 효과

남궁정 ( Jeong Namkung ),배수미 ( Su Mi Bae ),문란영 ( Lan Ying Wen ),오은경 ( Eun Kyeong Oh ),신재은 ( Jea Eun Shin ),김용욱 ( Yong Wook Kim ),김태응 ( Tae Eung Kim ),박태철 ( Tai Churl Park ),안웅식 ( Woong Shick Ahn ) 대한산부인과학회 2009 Obstetrics & Gynecology Science Vol.52 No.2

목적: As2O3는 다양한 고형암에서 혈관손상을 통하여 항종양 효과를 유도하는 것으로 알려져 있다. 본 연구에서는 자궁경부암동물 모델에서 새로운 비소 화합물인 As4O6와 As2O3를 비교하여 종양 내 혈관폐쇄 현상을 확인하고자 하였다. 연구 방법: HPV 16-E6/E7 유전자를 가지고 있는 TC-1 종양세포를 이용한 자궁경부암 동물모델에서 종양의 부피가 200~250 ㎣에 이르면 PBS, As2O3 and As4O6를 복강 내 투여하였다. 비소 화합물을 주사한 후 종양의 크기는 2~3일 간격으로 측정하고, 종양내혈관 폐쇄현상은 Evans blue 추출법과 Hoechst 33342 염색법 등 2가지 방법을 이용하여 측정하였다. 또한 헤마토실린-에오신 염색을 통하여 조직학적 변화를 관찰하였다. 결과: As2O3와 As4O6 주사 후, 종양 성장은 대조군에 비해 효과적으로 지연되었으며, As4O6 처리군이 가장 우수한 항종양 효과를 나타내었다. 또한 비소화합물의 투여는 종양내의 거대한 세포괴사와 혈관폐쇄 현상을 유도하였다. 결론: TC-1 세포주를 이식한 고형암 동물모델에서 As4O6의 처치는 혈관폐쇄 효과에 의한 항종양 효과를 나타내었다. Objective: Arsenic trioxide (As2O3) is known to have potent anti-vascular activity and significantly suppress solid tumor growth. The present study was conducted to investigate the vascular shutdown effects of a novel arsenic compound, tetraasrsenic oxide (As4O6), in comparison with As2O3 using cervical cancer animal model. Methods: Mice tumor challenge model was used C57BL/6 mice transplanted with TC-1 cells. After the growth of tumors was reached up 200~250 ㎣, mice were divided into 3 groups randomly for control and treatment of either As2O3 or As4O6. As2O3 and As4O6 was treated by i.p. injection. The tumor size was caliperated in twice for weeks and anti-vascular effect were assessed by Evans blue extraction assay and Hoechst 33342 staining. In tumor tissue, histopathological feaure was obserevd by hematoxylin and eosin (H&E) staining. Results: In mice treated with either As2O3 and As4O6 (i.p.), both of As2O3 and As4O6 was significantly suppressed the tumor growth compared with control group. Moreover, effect of As4O6 is more pronounced. These tumor growth inhibition is led to the massive necrosis and vacular shutdown in tumor tissue. Conclusion: This study suggests that As4O6 may have potential anticancer activity via vascular shutdown in C57BL/6 mice transplanted with TC-1 cells.

임신 중기에 발생한 림프구성 뇌하수체염

고현선 ( Hyun Sun Ko ),이희중 ( Hee Joong Lee ),이근호 ( Keun Ho Lee ),장은덕 ( Eun Deok Chang ),강석구 ( Seok Gu Kang ),신종철 ( Jong Chul Shin ),박태철 ( Tai Churl Park ) 대한주산의학회 2005 Perinatology Vol.16 No.4

Lymphocytic hypophysitis, a rare inflammatory disorder in the pituitary gland that is often associated with pregnancy, is caused by autoimmune destruction of the pituitary gland evidenced by diffuse lymphocytic infiltration in the pituitary gland. We report a case of lymphocytic hypophysitis in a 38-year-old woman presenting with aggravating headache and sudden visual loss during pregnancy, which was misdiagnosed as pituitary adenoma before neurosurgery. Magnetic resonance imaging, with enhancement, showed an expanding 2.5×2.0×1.7 cm sized sellar mass. The patient underwent transsphenoidal surgery with a preoperative diagnosis of pituitary macroadenoma. A histological examination revealed diffuse lymphocytes and fibrosis of adenohypophysis with atrophy, which are characteristic of lymphocytic hypophysitis. We report a case of lymphocytic hypophysitis occurred during pregnancy with a brief review of literatures.