단기배양한 중피세포의 면역세포화학적 연구

전호종,이미자,이미숙,정유경,이영미,최형호,Jeon, Ho-Jong,Lee, Mi-Ja,Lee, Mi-Sook,Jeong, Yu-Kyung,Lee, Young-Mi,Choi, Hyung-Ho 대한세포병리학회 1995 대한세포병리학회지 Vol.6 No.2

Reactive humsn mesothelial cells were examined by immunocytochemical stain with intermediate filaments (cytokeratin [CK1, CK7, CK8, CK18, CD19), vimentin, desmin, actin), epithelial membrane antigen, carcinoembryonic antigen (CEA), MHC class II antigen (HLA-DR), LeuM-1 (CD15), $\alpha1-antitrypsin$(ACT), $\alpha1-antichymotrypsin$ (ACHT), CD68(KP-1) and FcyRIII(CD16). The mesothelial cells were isolated from patients with liver cirrhosis and pleural effusion, and short-term cultured in RPMI 1640 media containing 10% heat inactivated fetal calf serum and 1% identical supernatant fluid of the patients' transudates. The results obtained are as follows 1. The cultured-reactive mesothelial cells were positive for the protein of cytoskeleton such as cytokeratin and vimentin, but negative for desmin and actin. The resting mesothelial cells showed positive reactions for cylokeratin, but negative for vimentin, desmin and actin. 2. The primary antibodies to the cytokeratin were strongly reactive for CK1, CK8 and CK18 but negative for CK7 and CK19 in both reactive and resting mesothelial cells. 3. Resting mesothelial cells showed negative reactions for CEA, but strong positive reactions in cultured-reactive mesothelial cells. 4. The markers for the monocytes/histiocytes(CD11b, CD14, CD16, CD68, Iysozyme and $\alpha1-antitrypsin$ and $\alpha1-antichymotrypsin$) were nonreactive in resting mesothelial cells, but lysozyme and $\alpha1-antitrypsin$ were weakly reactive in reactive and proliferative mesothelial cells. 5. MHC Class II molecule(HLA-DR antigen) was negative in both resting and reactive mesothelial cells. These results suggest that the short-term cultured, reactive mesothelial cells show a newly aberrant expression of the vimentin and calcine-embryonic antigen. The reason of the aberrant expression of the intermediate filament and oncofetal antigen in reactive and proliferative mesothelial cells should be further evaluated.

체액도말에서의 AgNOR수의 유의성 - 반응성 중피세포와 악성세포의 감별 -

백승삼,홍은경,장세진,박문향,이중달,Paik, Seung-Sam,Hong, Eun-Kyung,Jang, Se-Jin,Park, Moon-Hyang,Lee, Jung-Dal 대한세포병리학회 1997 대한세포병리학회지 Vol.8 No.2

To distinguish reactive mesothelial cells from malignant cells in body fluid, we applied silver staining of nucleolar organizer regions(AgNORs) to ethanol fixed cytologic preparations. Fifty aspirated samples of benign(22 cases) and malignant(26 cases) body fluids were studied using the one step silver staining method. Two cytologically atypical samples were also included in the study. In malignant cases the mean AgNOR count was $3.56{\pm}0.81$, while in benign cases the mean AgNOR count was $2.02{\pm}0.33$. The difference of AgNOR counts between these two groups were statistically significant(p<0.001). The mean of atypical cases was 2.91. Both were diagnosed as malignant in follow-up cytology. In malignant effusions, there is statistically significant difference in AgNOR counts between cells forming complex papillae or clusters and singly scattered cells(p<0.05), $3.29{\pm}0.95\;and\;3.83{\pm}0.55$, respectively. We concluded that AgNOR count appears to be useful as a diagnostic tool especially when the cytologic differentiation is difficult.

Usefulness of E-Cadherin Expression in Malignant Effusion

임성직,김교영,김윤화,박용구,양문호,원남희,이주희,Lim, Sung-Jig,Kim, Gou-Young,Kim, Youn-Wha,Park, Yong-Koo,Yang, Moon-Ho,Won, Nam-Hee,Lee, Ju-Hie The Korean Society for Cytopathology 1999 대한세포병리학회지 Vol.10 No.2

체강액 내의 악성 종양세포와 중피세포의 구분이 종종 어려우나 환자의 치료나 종양의 임상기 결정에 큰 영향을 미치기 때문에 정확히 감별하는 것이 매우 중요하다. E-cadherin은 상피세포에서 표현되는 유착단백질이다. 본 연구에서는 체강액의 세포학적 검사에서 악성세포의 표지자로서 E-cadherin의 유용성을 알아보기 위하여 세포검사후 조직검사로 진단을 확인한 33예를 대상으로 체강액으로부터 만든 세포 블록에 대하여 E-cadherin에 대한 면역세포화학염색을 시행하였다. 33 예의 세포학적 진단은 선암종 25예, 비정형세포 8예였다. 선암종으로 진단하였던 25 예중 21예(84%)에서 E-cadherin 에 양성이었다. 비정형세포라고 진단하였던 8예중 6예에서 음성이었으며 양성으로 염색된 2예는 조직학적 검사로 전이성 암종임을 확인하였다. 반응성 중피세포나 염증세포는 모두 음성이었다. 민감도와 특이도는 각각 84%와 75%였다. 결론적으로 E-cadherin은 체강액에서 악성 종양세포와 반응성 중피세포와의 구분에 유용한 보조적인 표지자이다. The usefulness of E-cadherin immunostaining as a marker of malignancy in the body fluids was investigated in the present study. Thirty-three histologically proven cases of cell blocks from the pleural, peritoneal, and pericardial fluids were studied by immunocytochemistry for E-cadherin antibody using LSAB method. These cases were cytologically diagnosed as adenocarcinoma (25 cases) and atypical cells (8 cases). Tumor cells showed strong positive membranous staining for E-cadherin antibody in 21 out of 25 cases (84%) of adenocarcinoma. E-cadherin staining was not found in 6 of 8 cases of suspicious maligancy. The sensitivity and specificity were 84% and 75%, respectively. Reactive mesothelial cells and Inflammatory cells scattered were all negative. In conclusion, E-cadherin is an useful adjunctive marker to distinguish reactive mesothelial cells from the carcinoma cells in the body fluids.

포도당분해산물이 사람 복막중피세포 활성화에 미치는 영향

송재숙,이경림,하헌주 이화여자대학교 약학연구소 2008 藥學硏究論文集 Vol.- No.17

상용 복막투석액에 함유된 고농도의 포도당과 포도당 분해산물(glucose degradation products: GDP)이 복막의 비후, 복막 투과성의 증가 및 한외여과 부전과 같은 복막의 구조적, 기능적 변화를 초래하리라 추정되고 있다. 본 연구에서는 GDP 성분이 사람 복막중피세포 활성화에 미치는 영향을 검색하였고 또 이때 ROS와 PKC가 관여하는지를 검색하였다. 혈청이 배제된 M199 배양액으로 성장을 동일화시킨 사람 복막중피세포를 GDP인 methylglyoxal(MGO), acetaldehyde, 그리고 3,4-dioxyglucosone-3-ene(3,4-DGE)으로 48시간 동안 자극하였고, 복막의 투과성에 대한 지표로서 혈과내피성장인자(vascular endothelial growth factor: VEGF)를, 섬유화의 지표로서 fibronectin과 heat shock protein 47(hsp47)의 단백을 정량하였다. 활성산소족(reactive oxygen species: ROS)과 protein kinase C(PKC)의 관여여부는 각각 항산화제 N-acetylcystein(NAC)과 PKC 억제제 calphostin C의 억제 효과로 검색하였다. MGO는 대조군과 비교하여 VEGF분비를 1.9배, fibronectin 분비를 1.5배 그리고 hsp47 표현을 1.3배로 유의하게 증가시켰다(p<0.05). MGO에 의한 VEGF 상향 조절은 calphostin C와 NAC에 의하여 유의하게 억제되었다. 사람 복막중피세포에서 VEGF 분비는 acetaldehyde에 의하여 증가하였으나 3,4-DGE에 의하여 억제되었고, fibronectin 분비와 hsp47 표현은 acetaldehyde나 3,4-DGE에 의하여 영향을 받지 않았다. 이상을 종합할 때, ROS 생산과 PKC 활성화가 상용투석액내 함유된 MGO에 의한 점진적인 복막의 투과성 증가, 세포외기질 축적 그리고 복막 섬유화를 유발하는 주된 신호체계로서 이를 선택적으로 억제함으로써 복막의 기능을 유지할 수 있을 것으로 생각된다. Both high glucose and glucose degradation products (GDP) have been implicated in alterations of peritoneal membrane structure and function during long-term peritoneal dialysis (PD). The present study examined the role of GDP including methylglyoxal (MGO), acetaldehyde, and 3,4-dideox-yglucosone (3,4-DGE) in HPMC activation with respect to membrane hyperpermeability or fibrosis. The role of reactive oxygen species (ROS) and activation of protein kinase C (PKC) in GDP-induced HPMC activation were also examined. Using M199 culture medium as control, growth arrested and synchronized HPMC were continuously stimulated by MGO, acetaldehyde, and 3,4-DGE for 48 hours. Vascular endothelial growth factor (VEGF) was quantified as a marker of peritoneal membrane hyperpermeability and fibronectin and heat shock protein 47 (hsp47) as markers of fibrosis. Involvement of ROS and PKC was examined by the inhibitory effect of N-acetylcystein (NAC) or calphostin C, respectively. MGO significantly increased VEGF (1.9-fold), fibronectin (1.5-fold), and hsp47 (1.3-fold) secretion compared with control M199. NAC and calphostin C effectively inhibited MGO-induced VEGF upregulation. Acetaldehyde stimulated and 3,4-DGE inhibited VEGF secretion. Fibronectin secretion and hsp47 expression in HPMC were not affected by acetaldehyde or 3,4-DGE. In conclusion, MGO upregulated VEGF and fibronectin secretion and hsp47 expression in HPMC, and PKC as well as ROS mediate MGO-induced VEGF secretion by HPMC. This implies that PKC activation and ROS generation by GDP may constitute important signals for activation of HPMC leading to progressive membrane hyperpermeability and accumulation of extracellular matrix and eventual peritoneal fibrosis.

식품 및 환경, 기타 포도당분해산물이 사람 복막중피세포 활성화에 미치는 영향

송재숙 ( Jae Sook Song ),이경림 ( Kyung Lim Lee ),하헌주 ( Hun Joo Ha ) 한국미생물생명공학회 2005 한국미생물·생명공학회지 Vol.33 No.4