멜라닌세포성 피부병변에서 AgNORs의 단순 계수법과 컴퓨터 영상분석법의 비교연구

이성열(Sung Yul Lee),오칠환(Chil Hwan Oh) 대한피부과학회 1993 大韓皮膚科學會誌 Vol.31 No.6

Background : Nucleolar organizer regions(NORs) are loops of DNA that transcribe ribosomal RNA; they can be easily identified in paraffin section using silver method. Recently, the application of NORs to the study of various types of benign and malignant proliferation has suggested that the number of NORs per nucleus can provide an indication of degree of cellur maligriancy. Many authors indicated a significant overlapping of NORs counts between benign and malignant proliferation. Objective : The purpose of this study is to discriminate benign m laocytic nevi from malignant melanoma effectively, thus we use simple enumeration and computerized image analysis of AgNORs in melanocytic skin lesions. Method : NORs were investigated on silver stained histologicec ion of 18 malignant melanoma (MM), 19 acquired nevi(AN), 21 congenital nevi(CN). For each case, 100 nucleus sample were cuantified using image analysis system(AIC Inc., Roswell, GA). There is no consensus at present as to the best criterion for quantifying AgNORs proteins in melanocytic skin lesion. We prefer to quantify a large number of parameters. The mean value or standard deviation(SD), each calculated for a batch of 100 cells, are average again(mean+SD), over the 58 cases for six morphological criteria mean number of AgNORs per nucleu, mean ratio of AgNORs area per ruc eus area, mean area of largest AgNORs, mean ratio of largest AgNORs area per nucleus area, mean nucleus area per a AgNOR, and coefficient of variation of nucleus area. Results : 1. All of the six parameters show significant difference between benign melanocytic nevi and malignant melanoma. But., there is no significant difference between acquired nevi and congenital nevi. 2. In stepwise discriminate analysis, we discriminate benign melanoctic nevi from malignant melanoma effectively using combined two parameters(number of AgNORs and mean ratio of AgNORs area per nucleus area). But there is some overlapping between malignant melanoma and ber ign melanocytic nevi using each six parameters. 3. In metastatic and non-metastatic malignant melanoma, six param ters have not shown significant difference. Conclusion : Study of AgNORs in the image analysis system is a very useful tool to differentiate malignant melanoma from benign melanocytic nevi. (Kor J Dermatol) 1993;31(6):866-876)

9,10-Dimethyl-1,2-benzanthracene으로 유도된 햄스터 협낭 발암과정 중 AgNORs과 증식성 핵항윈(PCNA)의 악성도 측정 표지자로서의 유의성 평가

홍삼표,송명순,홍성두,이재일,임창윤 대한병리학회 1998 Journal of Pathology and Translational Medicine Vol.32 No.5

대장 및 직장의 침윤성 선암종에서 AgNORs수와 환자의 생존율과의 관계

강구,남은숙,신형식,김순란,박영의,이재정,박철재,이민철 대한대장항문학회 1997 Annals of Coloproctolgy Vol.13 No.1

방광이행세포암에서 PCNA 및 AgNORs 발현양상

박병곤,이상용,노미숙,나서희,홍숙희 대한병리학회 1994 Journal of Pathology and Translational Medicine Vol.28 No.5

자궁내막증식증과 암종에서의 AgNOR, PCNA 및 유세포분석기를 이용한 DNA배수체 분석

성순희 梨花女子大學校 醫科大學 醫科學硏究所 1998 EMJ (Ewha medical journal) Vol.21 No.1

자궁내막의 증식증과 암종을 각 조직학적 등급별로 나누어 세포역동학적인 측면에서 이들의 상관관계를 규명하고자 파라핀포매조직을 이용하여 AgNOR 염색과 PCNA(proliferative cell nuclear antigen)에 대한 면역조직화학적 검색을 통해 형태계측학적인 방법으로 지수를 구하여 각각을 비교하여 보았으며 유세포 분석기를 이용하여 DNA배수체를 분석하였다. 내막증식증의 각 유형별 및 내막암종의 등급별 AgNOR수와 PCNA 지수는 정상, 단순형 증식증, 복합형 증식증, 비전형 증식증, 내막암종의 Ⅰ등급, Ⅱ등급, Ⅲ등급의 순으로 일련으로 증가하여 이미 알려진 각 유형의 악성 잠재력과 유의한 상관관계를 보였다. 또한 내막증식증중 비전형 증식증이 다른 유형의 증식증에 비하여 의미있게 증가하였고 오히려 Ⅰ등급의 내막양 암종과 유사하여 세포역동학적 측면에서 두 질환은 유사하게 간주되어야 할 것이다. 내막양 암종에서는 분화가 나쁜 Ⅲ등급이 Ⅰ,Ⅱ등급에 비해 의미있게 증가하여 분화도와 이들 증식지수간에 상관관계가 있는 것으로 해석된다. 또한 내막암종에서 분화도가 나쁠수록 비배수체가 증가하는 경향이었다. Endometrial hyperplasia(EH) and endometrial carcinoma(EC) very in their biologic potential, which may be correlated with the histologic grade. Evaluation of cellular kinetics, which may prove to be another measure of predicting biologic behavior. Accessments of AgNORs and PCNA(proliferative cell nuclear antigen) indeces in 33 cases of EH including 16 cases of simple hyperplasia(SH), 8 of complex hyperplasia(CH), and 9 of atypical hyperplasia(AH), and 28 of were as follows :Ⅰ, 12 of grade Ⅱ, and 6 of grade Ⅲ were performed. The results were as follows : 1. AgNOR counts per glandular cells(Mean SD) were 2.7±0.2 in normal proliferative and 2.3±0.2 in secretory endometrium, and increased to 3.2±0.3 in SH, 3.5±0.3 in CH, to 5.4±0.4 in AH, and finally 6.9±0.5 in endometrioid carcinoma(grade Ⅰ: 5.8±0.7, grade Ⅱ: 6.7±0.6, grade Ⅲ: 8.4±0.9). 2. PCNA indeces(percentages of nuclear positive cells of total cells of glands) were 16±14.2 in normal proliferative endometrium and 12±8.1 in secretory endometrium, and increased to 18.4±14.7 in SH, 21.6±17.8 in CH, 36.4±27.4 in AH, and finally 42.1±31.3 in EC(grade Ⅰ: 38.3±23.2,grade Ⅱ: 39.4±25.4, and grade Ⅲ: 58.4±35.3). 3. DNA aneuploidy was detected in 4 cases of EC(40%), and tended to be more frequently found in poorer histologic grade. This data suggest that cell kinetic evaluation of EH and EC using AgNORs and PCNA is well correlated with histologic grade. And with the aspect of biologic potential, AH could be regarded as well differentiated EC.

신세포암종에 있어서 핵등급과 AgNORs수 및 PCNA 표지지수 사이의 상관관계

이혜진,한영임,서강석,이선경 대한병리학회 1996 Journal of Pathology and Translational Medicine Vol.30 No.2

성상세포종의 악성도와 Ki-67, PCNA, AgNORs 및 p53 단백 발현의 상관관계

김동자,임재원,손윤경 대한병리학회 1999 Journal of Pathology and Translational Medicine Vol.33 No.1

수막종에서 스테로이드 호르몬 수용체, PCNA 및 AgNORs의 진단적 의의

김상표,권건영 계명대학교 의과학연구소 2000 계명의대학술지 Vol.19 No.2

We have reviewed immunohistochemically 80 cases with meningiomas in order to determine if correlation exist that might be useful as the diagnostic and prognostic factors on these tumors. Intracranial meningiomas obtained from 80 patients(55 female and 25 male, mean age 51.2±14.8, range 7-83 years) were examined for steroid hormones receptors, proliferating cell nuclear antigen(PCNA) and nucleolar organizer regions(AgNORs). Eighty intracranial meningiomas were classified by 66 benign, 4 recurrent, 7 atypical, and 3 malignant. Progesterone receptors(PRs) and estrogen receptors(ERs) status was scored as 0(0% nuclei positive), 1(<1%), 2(1-9%), 3(10-49%), or 4(≥50%). Nuclear staining for the PRs was found in 62 cases (77.5%), which was showed 50 of 66 in benign, 3 of 4 in recurrent, 6 of 7 in atypical and 3 of 3 in malignant. They showed statistically significant correlation(p<0.05) between tumor grading and PRs staining score. PRs staining score of recurrent four meningiomas also was lower(p<0.05) than those of non-recurrent benign meningiomas. There was no evidence of nuclear ERs staining in all cases. Immunostaining for PCNA revealed higher labelling indices in malignant meningiomas than that in benign, recurrent and atypical meningiomas. The mean number of nuclei for AgNORs tended to significantly increased in atypical and malignant meningiomas than that in benign and recurrent meningiomas. Therefore, PRs, PCNA and AgNORs are a useful adjunct in identifying a diagnostic and prognostic utility on meningiomas.

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.

Flowcytometric quantitative analysis and immunoblotting patterns of the c-myc oncoprotein in hepatocarclnogenesis

정자영(Ja Young Jeong),이영순(Yong Soon Lee) 한국예방수의학회 1998 예방수의학회지 Vol.22 No.3

본 연구는 화학적 간발암화 과정에서 c-myc 종양단백의 immunoblotting 패턴과 유세포 측정기를 이용한 정량분석을 수행하였다. 또한 종양성 간세포의 증식성 평가를 위하여 핵당 silver-binding nucleolar organizer regions(AgNORs)를 조사하였다. 실험군은 세 군으로 나누었으며, Ⅰ군은 대조군, Ⅱ군은 암 유발 물질 투여군으로, 2,2’-dihydroxy-di-n-propylnitrosamine(DHPN, 0.1% in d.w.)과 N-ethyl-N-hydroxyethylnitrosamine(EHEN, 0.15% in d. w.)을 1주 동안씩 각각 처치하였다. 암유발 및 촉진군으로 Ⅲ군은 암유발 물질 투여후, 촉진을 위하여 3’-methycholanthracene(3’-MC, 10mg/kg, i.p., twice a week)와 DL-serine (0.05% in d.w.)을 5주 동안 처치하였다. 모든 동물은 8, 12 그리고 20주에 희생시켜 조사하였다. 면역형광 염색은 붉은 색의 핵에 녹색의 둥근 점이 산재되어 관찰되었다. Dual-parametric FCM으로 분석한 c-myc 퍼센트는 Ⅰ군에서 18%, Ⅱ군에서 21~23% 그리고 Ⅲ군에서는 24~26%로 조사되었으며, 부검시간과 처치에 따른 유의차는 인정되지 않았다. c-myc 종양단백의 immunoblotting 조사에서는, 발암물질을 처치하지 않은 Ⅰ군에서 68 kDa의 강한 하나의 밴드가 관찰되었다. 그러나 발암물질 처치군인 Ⅲ군에서는 68 kDa 밴드 이외에 65 kDa의 밴드가 새롭게 나타났다. Ⅱ군의 AgNORs의 평균수는 암병변에서 정상부위에 비하여 8주 (p<0.05), 12주(p<0.01)에서 유의성 높게 나타났다. Ⅲ군에서도 역시 전암병변에서 8주 (p<0.05), 12주 (p<0.05) 그리고 20주(p<0.01)에서 유의성 높게 관찰되었다. 결론적으로 유세포 측정기로 측정한 c-myc 종양단백의 함량 측정방법은 간편하고, 단순하며 신속한 방법이었다. 발암물질을 처치한 간에서 immunoblotting한 c-myc 종양단백은 68 kDa이 약해지고, 새롭게 65 kDa이 합성되어 나타났다. 따라서 발암물질에 의하여 새롭게 합성된 p65c-myc는 발암물질의 투여 결과이며, 발암물질에 의한 구조적 변성에서 기인된 것으로 사료된다. 한편 핵당 AgNORs의 평가는 종양성 간세포 또는 증식성 세포의 평가를 가능하게 해주는 것으로 조사되었다.