• Title/Summary/Keyword: Protein Expression

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The Study of CYFRA 21-1 and Epidermal Growth Factor Receptor Levels in Cancer Tissue of Bronchogenic Carcinoma Patients (폐암환자의 암조직내 CYFRA 21-1과 Epidermal Growth Factor Receptor의 측정치에 대한 연구)

  • 김대연;김송명
    • Journal of Chest Surgery
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    • v.30 no.9
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    • pp.854-861
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    • 1997
  • CYPRA 21-1 is known to be a cytokeratin 19 fragment, and it can be detected by using two specific monoclonal antibodies (KS 19-1 and BM 19-21) and can be clinically applied as a useful circulating tumor marker The epidermal growth factor receptor (EGF-R) expression was evaluated and characterized by its tyrosine protein kinase activity and by its ligand-stimulated autophosphorylation, a property shared with other peptide growth factor receptors. Autocrine or para'urine action was initiated by a growth factor, or by a transforming growth factor o, which had an extensive homology with EGP and which also stimulated tyrosine kinase activity on the EGF-R. The CYFRA 21-1 and the EGF-R levels in 30 patients with primary lung tumors were investigated. There were 24 patients with squamous cell carcinomas and 6 patients with adenocarcinomas. Specimen 5 mm3 in size were sampled at three different locations ; the main lesion, the boundary between the lesion and the unaffected tissue, and the unaffected tissue of the patients. The results were as follows 1. The CYPRA 21-1 concentration in the cancer boundary, the most malignant region,(348.6 : 89.9 ng/ml) was the lowest value. The CYFRA 21-1 concentration in unaffected tissue,(718.4$\pm$77.8 ng/ml) was higher than that in the main lesion. which had intact cellularity. 2. The EGF-R concentration in the main lesion was higher than that in the unaffected tissue, and the EGF-R concentration in a squamous cell cacinoma was higher than that in an adenocarcinoma. also, the EGF-R concentration in the cancer b undary was highest at stage 1, ll. The EGF-R concentration was higher in the main cancer lesion that in the unaffected tissue at stage 111, IV. 3. The CYFRA 21-1 was a cytoplasmic skeleton and the EGF-R was a cell-wall component; there was no correlation. In conclusion, CYFRA 21-1 was abundant in the cytoplasm but had a higher concentration in the unaffected tissue than in the main cancer lesion. The CYFRA 21-1 concentration of the tissue did not reflect the amount of cancer activity, the EGP-R was located in the cell membrane, the level of tissue that reflects cancer activity, so the main cancer lesion had a higher concentration than the unaffected tissue. CYFRA 21-1 is not a useful tumor maker at the tissue level. Because the EGF-R concentration re(looted the cancer activity, its a useful tumor marker for lung cancer.

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Identification of Oocyte-Specific Diva-Associated Proteins using Mass Spectrometry (Mass Spectrometry를 이용한 난자 특이적인 Diva와 상호작용하는 단백질의 동정)

  • Yoon, Se-Jin;Kim, Jung-Woong;Choi, Kyung-Hee;Lee, Sook-Hwan;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.3
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    • pp.189-198
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    • 2006
  • Objective: We previously described that Diva is highly expressed in matured metaphase II (MII) oocytes compared to immature germinal vesicle (GV) oocytes in mouse. We report here that the expression of Diva transcript as well as protein is oocyte-specific. To elucidate its physiological role in oocyte, the binding partner(s) of Diva has been identified by using immunoprecipitation (IP) followed by Mass Spectrometry. Methods: NIH/3T3 cells were transiently transfected for 24 h with either empty vector for control or FLAG-tagged mouse Diva construct, and IP was performed with anti-FLAG antibody. The immuno-isolated complexes were resolved by SDS-PAGE on a 12% gel followed by Coomassie Blue staining. For in-gel digestion, 15 bands of interest were excised manually and digested with trypsin. All mass spectra were acquired at a positive reflector mode by a 4700 Proteomics Analyzer (Applied Biosystems, Framingham, MA). Proteins were identified by searching the NCBI nonredundant database using MASCOT Peptide Mass Fingerprint software (Matrixscience, London). Results: Diva-associated complexes were formed in FLAG-tagged mouse Diva-overexpressed NIH/3T3 cells via IP using anti-FLAG-conjugated beads. Among the excised 15 bands, actin and actin-binding proteins such as tropomyosin, tropomodulin 3, and ${\alpha}$-actinin were identified. Binding between Diva and actin or tropomyosin was confirmed by IP followed by Western blot analysis. Both bindings were also detected endogenously in mouse ovaries, indicating that Diva works with actin and tropomyosin. Conclusions: This is the first report that immuno-isolated Diva-associated complexes are related to actin filament of the cytoskeletal system. When we consider the association of Diva with actin and tropomyosin, oocyte-specific Diva may play a role in modulating the cytoskeletal system during oocyte maturation.

IMMUNOHISTOCHEMICAL ASSAYS FOR THE EXPRESSION OF EPIDERMAL GROWTH FACTOR-SIGNALING PROTEINS IN ADENOID CYSTIC CARCINOMAS OF HUMAN SALIVARY GLANDS (타액선 선양낭성암종에서 상피성장인자 신호전달 단백의 발현에 관한 면역조직화학적 연구)

  • Park, Young-Wook;Kim, Jung-Hwan
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.6
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    • pp.499-510
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    • 2006
  • Malignant tumors of the human salivary glands may arise from major or minor salivary glands. Adenoid cystic carcinoma (ACC) is the second most common malignant neoplasm in the salivary glands. ACC is occasionally highly aggressive tumor that readily invades adjacent tissues and metastasize to distant organs at early stages of the disease. Although ACC tends to grow slowly, treatment outcome may be poor due to wide local infiltration, perineural or intraneural spread and a propensity for hematogenous metastasis. Therefore, knowledge of cellular and molecular characteristics that influence the growth, survival and metastasis of tumor cells, is important for new treatment strategies of salivary ACC. I determined expressions of epiderma growth factor (EGF)-signaling molecules using surgical specimens of human ACCs. Protein expressions of EGF, transforming growth $factor(TGF)-{\alpha}$, EGF receptor (EGFR), phosphorylated EGFR (pEGFR), and human EGF receptor (HER)-2 were assessed in 18 cases of salivary ACC by immunohistochemical staining. Adjacent normal salivary tissues and mucosal tissues, uninvolved by the malignant tumor, served as internal controls. Most of the tumors, especially ACC with a tubulocribriform pattern, were positive for EGF signaling molecules. The overall percentages of the 18 specimens expressing EGF, $TGF-{\alpha}$, EGFR, pEGFR, and HER2 were 50, 89, 61, 61 and 83% respectively. Moreover, tumor-associated endothelial cells and infiltrating immune-related cells in the stroma of ACC, also expressed these biomarkers. Taken together, EGF-signaling molecules are actively expressed in salivary ACC. Therefore, we suggest that these biomarkers can be molecular targets for new treatment strategies of salivary tumors.

Anti-Inflammatory Effect of Chondrus ocellatus Holmes Ethanol Extract on Lipopolysaccharide-induced Inflammatory Responses in RAW 264.7 Cells (Lipopolysaccharide로 유도된 RAW 264.7 세포와 마우스모델에 대한 진두발 에탄올 추출물의 항염증 효과)

  • Bae, Nan-Young;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Park, Ji-Hye;Park, Sun-Hee;Sung, Nak-Yun;Byun, Eui-Hong;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.268-277
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    • 2016
  • This study aimed to investigate the anti-inflammatory effect of the ethanol extract from Chondrus ocellatus Holmes (COHEE) in RAW 264.7 cells and in a mouse ear edema model, by measuring the production of lipopolysaccharide-induced inflammatory response mediators. There were no cytotoxic effects on the proliferation of macrophages treated with COHEE compared with the control. COHEE inhibited the production of nitric oxide and pro-inflammatory cytokines [interleukin (IL)-6, tumor necrosis factor-α, and IL-1β]. The extract also reduced the expression of inducible nitric oxide synthase, cyclooxygenase-2, nuclear factor-κB p65, and phosphorylated mitogen-activated protein kinase in a dose-dependent manner. In the croton-oil-induced ear edema model, COHEE decreased the formation of mouse ear edema at the highest dose compared with the control, and histological analysis revealed that the epidermal/dermal tissue thickness and mast cell numbers were reduced. Therefore, these results suggest that COHEE may be a promising topical anti-inflammatory therapeutic material through its action of modulating NF-κB and the MAPK signaling pathway.

A Cyclin-Dependent Kinase Inhibitor, p16^{INK4A}, Induces Apoptosis in The Human Cancer Cells. (Cyclin-dependent Kinase저해 단백질 p16^{INK4A}의 인체 암세포에서의 세포사멸 유도 활성)

  • 김민경;이철훈
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.72-77
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    • 2004
  • Previously, we synthesized a novel Cyclin-dependent kinase inhibitor, MCS-5A. Also, we investigated the involvement of cell cycle regulatory events during MCS-5A-mediated apoptosis in HL-60(+p16/-p53) cells with up-regulation of p16 protein expression. In contrast, apoptosis was not observed in A549(-p16/+p53) cells. Therefore we propose that $p16^{INK4A}$ is a key enzyme for inducing apoptosis. In the present studies, we have explored the mechanism of $p16^{INK4A}$ -mediated cytotoxicity and the role of p16.sup INK4A/ overexpression in the induction of apoptosis in human tumor cells. The tumor suppressor gene $p16^{INK4A}$ is known as a cyclin-dependent kinase inhibitor (CKI) and cell cycle regulator. We expressed wild type $p16^{INK4A}$ in pcDNA3.1 vector and then transfected into non-small cell lung cancer (NSCLC) cell expressing different statue of p16$^{INK4A}$, p53 gene〔A549(-p16/+p53), H1299(-p16/-p53) and HeLa(+pl6/+p53) cell line〕. TUNEL assay (including propidium iodide staining following transfection of these cell line with pcDNA3.1-pl6) indicate that p16$^{INK4A}$-mediated cytotoxicity was associated with apoptosis. This is supported by studies demonstrating an induction of caspase 3 cleavage due to the transfection of A549, H1299 and HeLa cells with pcDNA3.1-pl6. These results suggest that p16$^{INK4A}$ has a new function of inducing apoptosis which is not related with the function of tumor suppressor gene p53.

A Comparison of Illness Behavior among Patients with Somatoform Disorders, Depressive Disorders and Psychosomatic Disorders (신체형장애, 우울장애 및 정신신체장애 환자들간의 질병행동의 비교)

  • Koh, Kyung-Bong;Ki, Sun-Wan
    • Korean Journal of Psychosomatic Medicine
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    • v.5 no.2
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    • pp.185-194
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    • 1997
  • A comparison was made regarding illness behavior among patients with somatoform disorders, depressive disorders and psychosomatic disorders. The subjects consisted of out-patients with somatoform disorders(N=52), depressive disorders(N=52) and psychosomatic disorders(N=51). illness behavior was assessed by illness Behavior Assessment Schedule and the questionnaire about help-seeking behavior. The patients with somatoform disorders and psychosomatic disorders more often affirmed the presence of somatic disease, were more likely to have phobia of disease, had more preoccupation with ideas of disease and more frequently shopped around oriental clinics than the patients with depressive disorders. The patients with somatoform disorders more often attributed its cause to physical factors, less often attributed the origin of affective disturbance to psychological causes, showed Less depression and irritability, and were less likely to accept psychiatric treatment recommended by other physicians than depressive patients. The patients with somatoform disorders were more likely to report having been told that they suffered from a mild illness than those with psychosomatic disorders. The patients with somatoform disorders with psychological problems tended to inhibit expression of their emotion. Female patients with somatoform disorders more often affirmed the presence of psychological disorder and attributed its cause to psychological factors than male ones. These results suggest that in illness behavior, patients with somatoform disorders are different from depressive patients, whereas the former patients are similar to psychosomatic patients except the discrepancy between therapists and patients regarding evaluation of their symptoms. Thus, it is emphasized that first, therapists need to approach patients with somatoform disorders somatically with understanding of their underlying need to deny psychological problems, followed by either psychological or biopsychosocial approach.

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β-elemene Induces Caspase-dependent Apoptosis in Human Glioma Cells in vitro through the Upregulation of Bax and Fas/FasL and Downregulation of Bcl-2

  • Li, Chen-Long;Chang, Liang;Guo, Lin;Zhao, Dan;Liu, Hui-Bin;Wang, Qiu-Shi;Zhang, Ping;Du, Wen-Zhong;Liu, Xing;Zhang, Hai-Tao;Liu, Yang;Zhang, Yao;Xie, Jing-Hong;Ming, Jian-Guang;Cui, Yu-Qiong;Sun, Ying;Zhang, Zhi-Ren;Jiang, Chuan-Lu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10407-10412
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    • 2015
  • Background: ${\beta}$-elemene, extracted from herb medicine Curcuma wenyujin has potent anti-tumor effects in various cancer cell lines. However, the activity of ${\beta}$-elemene against glioma cells remains unclear. In the present study, we assessed effects of ${\beta}$-elemene on human glioma cells and explored the underlying mechanism. Materials and Methods: Human glioma U87 cells were used. Cell proliferation was determined with MTT assay and colony formation assay to detect the effect of ${\beta}$-elemene at different doses and times. Fluorescence microscopy was used to observe cell apoptosis with Hoechst 33258 staining and change of glioma apoptosis and cell cycling were analyzed by flow cytometry. Real-time quantitative PCR and Western-blotting assay were performed to investigated the influence of ${\beta}$-elemene on expression levels of Fas/FasL, caspase-3, Bcl-2 and Bax. The experiment was divided into two groups: the blank control group and ${\beta}$-elemne treatment group. Results: With increase in the concentration of ${\beta}$-elemene, cytotoxic effects were enhanced in the glioma cell line and the concentration of inhibited cell viability ($IC_{50}$) was $48.5{\mu}g/mL$ for 24h. ${\beta}$-elemene could induce cell cycle arrest in the G0/G1 phase. With Hoechst 33258 staining, apoptotic nuclear morphological changes were observed. Activation of caspase-3,-8 and -9 was increased and the pro-apoptotic factors Fas/FasL and Bax were upregulated, while the anti-apoptotic Bcl-2 was downregulated after treatment with ${\beta}$-elemene at both mRNA and protein levels. Furthermore, proliferation and colony formation by U87 cells were inhibited by ${\beta}$-elemene in a time and does-dependent manner. Conclusions: Our results indicate that ${\beta}$-elemene inhibits growth and induces apoptosis of human glioma cells in vitro. The induction of apoptosis appears to be related with the upregulation of Fas/FasL and Bax, activation of caspase-3,-8 and -9 and downregulation of Bcl-2, which then trigger major apoptotic cascades.

Anti-Oxidant, Anti-Melanogenic, and Anti-Inflammatory Activities of Zanthoxylum schinifolium Extract and its Solvent Fractions (산초 추출물 및 분획물의 항산화, 미백 및 항염증 활성)

  • Jin, Kyong-Suk;Oh, You Na;Park, Jung Ae;Lee, Ji Young;Jin, Soojung;Hyun, Sook Kyung;Hwang, Hye Jin;Kwon, Hyun Ju;Kim, Byung Woo
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.371-379
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    • 2012
  • This study was designed to explore new nutraceutical and cosmetic resources possessing biological activities from the plant kingdom. To fulfill this purpose, we analyzed the anti-oxidative, anti-melanogenic, and anti-inflammatory activities of Zanthoxylum schinifolium extract (ZSE) and its solvent fractions using in vitro assays and cell culture model systems. Three kinds of ZSE treated with methanol, ethanol, and water exhibited potent anti-oxidative activities through DPPH radical scavenging capacity, and inhibited in vitro DOPA oxidation. Furthermore, Z. schinifolium methanol extract (ZSME) inhibited the ${\alpha}$-melanocyte stimulating hormone, which induces melanin contents in B16F10 cells. Its anti-melanogenic activity originates from the inhibition of tyrosinase enzyme activity and melanogenesis related protein expression. Moreover, lipopolysaccharide induced nitric oxide production in the RAW 264.7 cell line was also ameliorated by ZSME treatment in a dose dependent manner. Among the four solvent fractions of ZSME treated with dichloromethane, ethyl acetate, n-butanol, and water, three fractions, except water, showed significant anti-melanogenic and anti-inflammatory activities. Taken together, these results provide important new insights into Z. schinifolium, indicating that it possesses numerous biological activities such as anti-oxidative, anti-melanogenic, and anti-inflammatory activities. Therefore, it may well serve as a promising material in the field of nutraceuticals and cosmetics.

Anti-Inflammatory Effect of Sargassum patens C. Agardh Ethanol Extract in LPS-induced RAW264.7 Cells and Mouse Ear Edema (LPS로 유도된 RAW 264.7 cell과 마우스 귀 부종 모델을 통한 쌍발이 모자반 에탄올 추출물의 항염증 효과)

  • Kim, Min-Ji;Kim, Min-Ju;Kim, Koth-Bong-Woo-Ri;Park, Sun-Hee;Choi, Hyeun-Deok;Park, So-Yeong;Kim, Ji-Hyun;Jang, Mi-Ran;Im, Moo-Hyeog;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.45 no.2
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    • pp.110-117
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    • 2017
  • The anti-inflammatory effect of Sargassum patens C. Agardh ethanol extract (SPEE) was examined based on the lipopolysaccharide (LPS)-induced inflammatory response in this study. SPEE treatment was not cytotoxic to macrophages compared to the control. The production of NO was suppressed by SPEE by approximately 28% at $100{\mu}g/ml$, and levels of interleukin-6, tumor necrosis $factor-{\alpha}$, and $interleukin-1{\beta}$ decreased in a dose-dependent manner. In addition, the expression of inducible nitric oxide synthase, cyclooxygenase-2, and nuclear $factor-{\kappa}B$ was suppressed by SPEE treatment. In vivo, croton oil-induced mouse ear edema was attenuated by SPEE and the infiltration of mast cells into the tissue decreased. Based on these results, SPEE inhibits the release of LPS-induced pro-inflammatory cytokines and mediators, suggesting that SPEE is a potential agent for anti-inflammatory therapies.

P53 Overexpression and Outcome of Radiation Therapy in Head & Neck Cancers (두경부종양 환자에서 p53의 과발현과 방사선치료결과)

  • Kim In Ah;Choi Ihl Bhong;Kang Ki Mun;Jang Ji Young;Kim Kyung Mi;Park Kyung Shin;Young Shin Kim;Kang Chang Suk;Cho Seung Ho;Kim Hyung Tae
    • Radiation Oncology Journal
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    • v.17 no.1
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    • pp.1-8
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    • 1999
  • Purpose : Experimental studies have implicated the wild type p53 In cellular response to radiation. Whether altered p53 function can lead to changes in clinical radiocurability remains an area of ongoing study. This study was performed to investigate whether any correlation between change of p53 and outcome of curative radiation therapy in patients with head and neck cancels. Methods : Immunohistochemical analysis with a mouse monoclonal antibody (DO-7) specific for human p53 was used to detect to overexpression of protein in formalin fixed, paraffin-embedded tumor sample from 55 head and neck cancer patients treated with curative radiation therapy (median dose of 7020 cGy) from February 1988 to March 1996 at 51. Mary's Hospital. Overexpression of p53 was correlated with locoregional control and survival using Kaplan-Meier method. A Cox regression multi-variate analysis was peformed that included all clinical variables and status of p53 expression. Results : Thirty-seven (67.2$\%$) patients showed overexpression of p53 by immunohistochemical staining in their tumor. One hundred percent of oral cavity, 70$\%$ of laryngeal, 66.7$\%$ of oropharyngeal, 66.7$\%$ of hypopharyngeal cancer showed p53 overexpression (P=0.05). The status of p53 had significant relationship with stage of disease (P=0.03) and history of smoking (P=0.001). The overexpression of p53 was not predictive of response rate to radiation therapy. The locoregional control was not significantly affected by p53 status. Overexpression of p53 didn't have any prognostic implication for disease free survival and overall survival. Primary site and stage of disease were significant prognostic factors for survival. Conclusions : The p53 overexpression as detected by immunohistochemical staining had significant correlation with stage, primary site of disease and smoking habit of patients. The p53 overexpression didn't have any predictive value for outcome of curative radiation therapy in a group of head and neck cancers.

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