• Title/Summary/Keyword: 면역조직세포화학 염색

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Detection of Apoptosis by M30 Monoclonal Antibody in Non-small Cell Lung Carcinomas (비소세포 폐암에서 단클론항체 M30를 이용한 세포자멸사 측정)

  • Kim, Gwang-Il;Lee, Gun;Lim, Chang-Young;Lee, Hyeon-Jae
    • Journal of Chest Surgery
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    • v.40 no.2 s.271
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    • pp.114-121
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    • 2007
  • Background: Apoptosis plays a crucial role in carcinogenesis, as well as in development and tissue homeostasis. Terminal deoxyribonucleotidyl transferase mediated neck end labelling (TUNEL) and in situ nick end labelling (ISEL) have been used to investigate the apoptosis in tissues. Since the introduction of the M30 monoclonal antibody to overcome drawbacks of TUNEL and ISEL, the apoptosis in various tumors, with the exception of pulmonary carcinomas, has been studied. In this study, attempts were made to examine the correlation of apoptosis in non-small cell carcinomas, using both M30 and the expression of p53 protein, with the clinicopathological factors. Material and Method: Forty five patients with surgically resected non-small cell carcinomas were included. Immunohistochemical staining with M30 and p53 monoclonal antibody were peformed, and their expressions compared with the clinicopathological features. The overall survival time and recurrence-free survival time were calculated, and the factors influencing the survival time analyzed using a univariate analysis. The effects of the expression stati of M30 and p53 on the risks of cancer related to both death and recurrence were evaluated using a multivariate analysis. Result: The p53 positive group had many more M30 positive cells than the p53 negative group (p53 positive group; $61.7{\pm}26.8$ cells vs. p53 negative group; $45.6{\pm}29.6$ cells, p=0.005) and significantly more p53 positive patients showing at least 10 positive cells (apoptotic index, $Al{\ge}1$) on M30 staining (p53 positive group; 52.4% (11/21) vs. p53 negative group 16,7% (4/24), p=0.025). In the univariate analysis, the survival times in relation to smoking (pack-year), performance status (PS) and Al showed significant differences. The multivariate analysis demonstrated the relative risk (R.R) of cancer death increased almost 7.5-fold (R.R 7.482; 95% Cl $1.886{\sim}29.678$; p=0.004) and the risk of recurrence almost 3,8-fold (R.R 3.795; 95% Cl: $1.184{\sim}12.158$; p=0.025) in the high Al (${\ge}1$) compared to the low Al (<1) group. There was no prognostic effect of p53 expression on the survival time or risk of cancer death and recurrence. Conclusion: In non-small cell lung carcinomas, M30 immunohistochemistry was an excellent method for analyzing apoptosis; the high apoptotic index could be an adverse prognostic predictive factor.

Immunocytochemical Localization of Metallothionein in Gastric Adenocarcinoma (위암 조직내 Metallothionein의 면역 세포화학적 연구)

  • Yang, Seung-Ha;Shin, Kil-Sang;Kim, Wan-Jong
    • Applied Microscopy
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    • v.32 no.4
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    • pp.411-419
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    • 2002
  • Metallothionein (MT) is a family of ubiquitous, low molecular weight ($6,000{\sim}7,000D$), cysteine-rich ($30{\sim}35%$) inducible protein with a high affinity to metal ions and has no aromatic amino acids and histidine. Some of the known functions of MT include detoxification of heavy metals and alkylating agents and neutralization of free radicals. Also, this protein has been reported to involve in tumor pathophysiology and therapy resistance. MT expression may affect a number of cellular processes including gene expression, apoptosis, proliferation and differentiation. Many reports on the physiological and biochemical properties of MT have been published, but ultrastructural reports on the localization of MT in human gastric cancer tissues are extremely rare. The present study was undertaken to examine the ultrastructural features and the localization of MT within the gastric adenocarcinoma. Ultrastructures of gastric cancer cells were characterized by the high nuclear cytoplasmic ratio, the interdigitation between cells, the irregular nucleus containing much heterochromatin and the wide distribution of free ribosomes in the cytoplasm. Immunohistochemical reaction for MT was prominent in the gastric adenocarcinoma. And the immunogold labellings were more prominent within the nucleus than the cytoplasm. Particularly, immunogold particles were numerously seen at nulcleolus or nucleolar associated heterochromatin. These results suggest that MT expression by gastric cancer cells is associated with cell proliferative activity and is possibly synthesized in the cytoplasm, and then the protein is transported into the nucleus to participate in any transcriptional steps.

Immunohistochemical Detection of the Grolwth Hormone-like Substance in Sparganum of Spirometra erinacei (고충(Sparganum)에서 성장호르몬 유사물질의 면역조직화학적 검출)

  • 김명옥;최완성김창환
    • The Korean Journal of Zoology
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    • v.35 no.2
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    • pp.173-182
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    • 1992
  • 흰쥐에 Spirometro rrinoce측 제3기 유충(고충 sparganum)이 감염되었을때 유충에서 생성된 성장호르몬 유사물질이 흰쥐 성장에 미치는 영향을 조사하기 위하여, 감염 기일이 경과에 따라 흰쥐의 뇌하수체와 유충의 체내에서 성장호르몬 분비세포를 면역조직화학염색으로 검색하였다. 유충의 표피와 표피성 근육층 및 근질 근육층에서 면역반응성을 띄는 성장 호르몬이 동정되었으며 감염시기의 경과에 따라 성장호르몬 분비세포의 수가 점차 증가하였다. 이와는 대조적으로 흰쥐의 뇌하수체에서는 성장호르몬 분비세포의 수가 감염 기일의 경과에 따라 정상대조군에 비하여 점차 감소 하였으며, 감염후 3개월이 경과되면 다시 증가하여 정상대조군의 수준으로 회복되었다. 또한 유충에 감염된 기일의 경과에 따른 횐쥐의 혈중 성장호르몬 농도변화를 dot-ELISA 방법으로 추정한 결깍 정상대조군의 성장호르몬의 양과 유의한 차이는 없었다. 결론적으로 유충의 체내에서 생성된 성장호르몬 유사물질이 전이숙주인 흰쥐의 성장을 유도할 것으로 사료된다.

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The Significance of p53 Expression in Serum and Tissue from Patients with Lung Cancer (원발성 폐암환자의 혈청 및 조직에서의 p53단백 표현)

  • Chang, Jung-Hyun;Sung, Sun-Hee
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.2
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    • pp.333-340
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    • 1998
  • Background: Lung cancer is the leading cause of cancer over the world. P53 alteration is by far the most common genetic defect in lung cancer. The mutation of p53 protein involves the loss of inhibitory function of p53 related tumor suppressor gene and resultant oncogenesis. The analysis of p53 alterations consists of immunohistochemical stain, PCR based assay, or serologic ELISA (enzyme-linked immunosorbent assay). Methods : Serum levels of p53 mutant protein were measured in 69 cases of lung cancer (adenocarcinoma n=29, epidermoid n=16, small cell n=13, large cell n=1, undifferentiated n=1, undetermined n=9) and 42 controls of respiratory disorders using ELISA. Immunohistochemical stain in tissue was performed using monoclonal antibody of p53 in lung cancer subjects. Results: Both serum p53s in nonsmall cell cancer ($0.28{\pm}0.44ng/ml$) and in small cell cancer ($0.20{\pm}0.14ng/ml$) were not different from controls ($0.34{\pm}0.20ng/ml$). Also there was no significant difference in serum p53 according to tumor stages. P53 immunohistochemical stain showed 50% positivity overall in lung cancer. There were no close correlation between serologic level and positivity of immunohistochemical stain. Conclusion: The serologic determination of p53 mutant protein is thought to have no diagnostic role in lung cancer. Immunohistochemical stain in lung cancer specimen shows 50% positivity.

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Effects of Opuntia ficus-indica Complexes on Blood Glucose and Pancreatic Islets Histology in Streptozotocin-induced Diabetic Rats (노팔천연복합물이 Streptozotocin으로 유발된 당뇨 쥐의 혈당 및 췌장조직에 미치는 영향)

  • Yoon, Jin-A;Kim, Je-Jung;Song, Byeng-Chun
    • Journal of the East Asian Society of Dietary Life
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    • v.22 no.3
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    • pp.334-340
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    • 2012
  • This study was carried out to investigate the effects of Opuntia ficus-indica complex (OF) on blood glucose, glucose tolerance, plasma insulin level and histopathological appearance of pancreatic islets in streptozotoxin (STZ)-induced diabetic rats. Thirty-two male Sprague-Daweley rats were divided into non-diabetic control (NC), diabetic control (DC), diabetic OF of 2% (OF-2) and diabetic OF of 5% (OF-5) and fed experimental diets for 3 weeks. Compared to the DC group fasting blood glucose levels in the OF-2 and OF-5 groups were significantly (p<0.05) reduced while fasting plasma insulin level in the OF-2 and OF-5 groups were significantly (p<0.05) increased. Glucose tolerance in the OF-2 and OF-5 groups were improved. Histopathological observation of pancreatic islets of the OF-2 and OF-5 groups showed hyperplasia which was very similar to NC. Numbers of ${\beta}$-cells in OF-2 ($47.81{\pm}0.92$) and OF-5 ($81.64{\pm}2.80$) were higher than numbers of ${\beta}$-cells in DC ($13.18{\pm}1.01$). These results imply that the intake of OF improves ${\beta}$-cell proliferation and prevents the death of ${\beta}$-cells in STZ-induced diabetic rats.

Differential Diagnosis of Pleural Mesothelioma and Metastatic Adenocarcinoma by Immunohistochemistry (면역조직화학염색법을 이용한 흉막의 악성중피종과 전이성 선암의 감별진단)

  • Ko, Kyung-Haeng;Park, Chang-Min;Rim, Myung-Soo;Kim, Yoo-Il;Jang, Il-Gweon;Hwang, Joon-Hwa;Lim, Sung-Chul;Kim, Young-Chul;Park, Kyung-Ok;Park, Chang-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.4
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    • pp.478-487
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    • 1999
  • Background : Differential diagnosis of pleural malignant mesothelioma from secondary metastatic adenocarcinoma is often difficult. A variety of pathologic techniques have been developed to make a differential diagnosis of carcinoma from mesothelioma. Immunohistochemistry detecting diverse antigenic substances such as CEA, Leu-M1, Bn-3, S-100 protein, vimentin, CK and EMA has been claimed to be of value as a panel in the differential diagnosis of adenocarcinoma from mesothelioma. The aim of this study was to investigate the suitable antibodies to distinguish mesothelioma from metastatic adenocarcinoma and establish candidate markers in a panel. Methods : Complete, one-hour immunohistochemical staining using antibodies against cytokeratin (CK), epithelial membrane antigen(EMA), S-100 protein, vimentin, B72-3, Leu-M1, and carcino-embryonic antigen(CEA) was applied to cell blocks from 7 mesotheliomas and 7 adenocarcinomas which were confirmed by electron microscopic and histpathologic methods. Results : All adenocarcinomas and 71.4% of mesotheliomas expressed the cytokeratin and EMA. S-100 protein and vimentin were expressed in 57.1% and 42.9% of mesotheliomas and 14.3% and 28.5% of adenocarcinomas, respectively. B72-3 was expressed in all adenocarcinomas, but in none of mesotheliomas. Leu-M1 was positive in 71.4% of the adenocarcinoma and 14.3% of the mesotheliomas. CEA was positive in all adenocarcinomas and 42.9% of mesotheliomas. Leu-M1 and B72-3 were coexpressed in 71.4% of adenocarcinomas but in none of mesothelioma. B72-3 and CEA were coexpressed in all adenocarcinomas, but in none of mesotheliomas. Conclusion : We concluded that B72-3 immunohistochemistry or panel staining of B72-3 and CEA could be recommanded for the differential diagnosis of pleural mesothelioma from metastatic adenocarcinoma.

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Immunohistochemical Localization of 36 and 29 kDa proteins in sparganum (면역조직염색법으로 관찰한 스파르가눔 층체에서의 36, 29 kDa 항원 단백질의 생성위치)

  • Kim, Lee-Su;Kong, Yoon;Kang, Shin-Yong;Cho, Seung-Yull
    • Parasites, Hosts and Diseases
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    • v.30 no.1
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    • pp.25-32
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    • 1992
  • Antigenic proteins of 36 and 29 kDa were localized in Spirometra mansoni plerocercoid (sparganum) immunohistochemically by avidin biotin complex (ABC) staining. When polyclonal antibodies such as BALB/c mouse serum immunized with crude saline extract of sparganum or confirmed sparganosis sera were reacted as pri-mary antibodies, the positive chromogen (3-amino, 9-ethylcarbazole) reactions were recognized at syncytial tegument, tegumental cells, muscle and parenchymal cells and lining cells of excretory canals. A monoclonal antibody(MAb) which was reacting to 36 and 29 kDa proteins in the extract of the worm was localized at the syncytial tegument and tegumental cells. The present results suggested that the potent antigenic proteins of 36 and 29 kDa in sparganum were produced at the tegumental cells and transported to the syncytial tegument.

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Laminin Expression in the Rat Lung Development (흰쥐 폐 발생시 Laminin의 발현에 대한 연구)

  • Chung, Ho-Sam;Park, Chul-Hong;Paik, Doo-Jin;Baik, Tae-Kyung;Kim, Won-Kyu;Youn, Jee-Hee;Suh, Yun-Kyung
    • Applied Microscopy
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    • v.31 no.1
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    • pp.71-83
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    • 2001
  • Laminin, a kind of multidomain glycoproteins, is mainly localized in the basement membranes of various tissues. It is known that laminin plays an important part in mammalian lung morphogenesis. The authors have undertaken this study to investigate the changes in the distribution of laminin, and to find out cells which synthesize laminin during the organogenesis and differentiation of the lung. The fetal and neoantal rats (Sprague-Dawley strain) were used as experimental animals. The immunohisto-chemical methods were employed for detection of laminin within the developing lung tissue and the immunegold cytochemical methods were performed for detection of cells which synthesize laminin according to each stage of development. The results are as follows; 1. During fetal life, strong immunoreactivity for laminin is maintained in the basement membranes of the blood vessels and the bronchioles, the extracellular matrix of the mesenchyme, and basal lamina of the alveolar septum in the fetal rat lung. 2. After birth, laminin immunoreactivity at the alveolar septum is gradually reduced. 3. During fetal life, laminin is mainly detected within the cytoplasm of the mesenchymal cells, the endothelial cells of blood vessels and the fibroblasts in fetal rat lung. 4. According to the differentiation of type I and type II pneumocyte after birth, laminin is detected within cytoplasm of the type I pneumocytes, type II pneumocytes and fibroblasts. It is consequently suggested that laminin is largely expressed in the developing lung and laminin may be also synthesized by the type II pneumonocytes at early newborn stages.

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Immunohistochemical Detection of Lymph Nodes Micrometastases in Patients of Pathologic Stage I Non-small-cell Lung Cancer (병리적 병기 1기의 비소세포폐암 환자에서 면역조직화학염색에 의한 림프절 미세전이 관찰)

  • Ryu, Jeong-Seon;Han, Hye-Seung;Kim, Min-Ji;Kwak, Seung-Min;Cho, Jae-Hwa;Yoon, Yong-Han;Lee, Hong-Lyeol;Chu, Young-Chae;Kim, Kwang-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.57 no.4
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    • pp.345-350
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    • 2004
  • Background : To evaluate the frequency and clinical significance of lymph node micrometastasis in patients of non-small-cell lung cancer pathologically staged to be T1-2,N0. Method : From consecutive 29 patients of non-small-cell lung cancer who received curative operation and routine systemic nodal dissection, we immunohistochemically examined 806 lymph nodes from mediastinal, hilar and peribronchial lesion. All slides were stained with hematoxylin and eosin staining for one section and with cytokeratin AE1/AE3 antibody for another consecutive section of same lymph node to find out micrometastasis. Results : In 806 lymph nodes examined, no tumor cell was seen on hematoxylin and eosin staining and micrometastic foci were shown to be on 0.37%(3) of 806 lymph nodes, in which were upper paratracheal, interlobar and peribronchial lymph node. These three positive stains constitute 10.3%(3) of the 29 patients with non-small-cell lung cancer. Nine patients died from disease progression(4), postoperative complication(3) and concomitant diseases(2). The four patients with disease progression did not show evidence of micrometastasis on their lymph node examination. Conclusion : The frequency of lymph node micrometastasis was in 0.37% of 806 lymph nodes examined. The study results might suggested that routine analysis of micrometastasis on the lymph node didn't give any clinical implication on patients with non-small-cell lung cancer.

The Signal Transduction Mechanisms on the Intestinal Mucosa of Rat Following Irradiation (방사선조사후 백서소장점막에서 발생하는 신호전달체계에 관한 연구)

  • Yoo Jeong Hyun;Kim Sung Sook;Lee Kyung Ja;Rhee Chung Sik
    • Radiation Oncology Journal
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    • v.15 no.2
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    • pp.79-95
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    • 1997
  • Purpose : Phospholipase C(PLC) isozymes play significant roles in signal transduction mechanism. $PLC-\gamma$ 1 is one of the key regulatory enzymes in signal transduction for cellular proliferation and differentiation. Ras oncoprotein, EGFR, and PKC are also known to be involved in cell growth. The exact mechanisms of these signal transduction following irradiation, however, were not clearly documented Thus, this study was Planned to determine the biological significance of PLC, ras oncoprotein, EGFR, and PKC in damage and regeneration of rat intestinal mucosa following irradiation. Material and Method : Sixty Sprague-Dawley rats were irradiated to entire body with a single dose of 8Gy. The rats were divided into S groups according to the sacrifice days after irradiation. The expression of PLC, ras oncoprotein, EGFR and PKC in each group were examined by the immunoblotting and immunohistochemistry. The histopathologic findings were observed using H&I stain, and the mitoses for the evidence of regeneration were counted using the light microscopy & PCNA kit. The Phosphoinositide(PI) hydrolyzing activity assay was also done for the indirect evaluation of $PLC-\gamma$ 1 activity. Results: In the immunohistochemistry , the expression of $PLC-{\beta}$ was negative for all grøups. The expression of $PLC-{\gamma}1$ was highest in the group III followed by group II in the proliferative zone of mucosa. The expression of $PKC-{\delta}1$ was strongly positive in group 1 followed by group II in the damaged surface epithelium. The above findings were also confirttled in the immunoblotting study. In the immunoblotting study, the expressions of $PLC-{\beta}$, $PLC-{\gamma}1$, and $PKC-{\delta}1$ were the same as the results of immunohis-tochemistry. The expression of ras oncoprctein was weakly positive in groups II, III and IV. The of EGFR was the highest in the group II, III, follwed by group IV and the expression of PKC was weakly positive in the group II and III. Conclusion: $PLC-{\gamma}1$ mediated signal transduction including ras oncoprotein, EGFR, and PKC play a significant role in mucosal regeneration after irradiation. $PLC-{\delta}1$ mediated signal transduction might have an important role in mucosal damage after irradiation. Further studies will be necessary to confirm the signal transduction mediating the $PKC-{\delta}1$.

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