• Title/Summary/Keyword: Cancer cells

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Plasminogen Activator Inhibitor Type 1 (PAI-1) A15T Gene Polymorphism Is Associated with Prognosis in Patients with EGFR Mutation Positive Pulmonary Adenocarcinoma

  • Lim, Ju Eun;Park, Moo Suk;Kim, Eun Young;Jung, Ji Ye;Kang, Young Ae;Kim, Young Sam;Kim, Se Kyu;Shim, Hyo Sup;Cho, Byoung Chul;Chang, Joon
    • Tuberculosis and Respiratory Diseases
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    • v.75 no.4
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    • pp.140-149
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    • 2013
  • Background: Plasminogen activator inhibitor type 1 (PAI-1), an important regulator of plasminogen activator system which controls degradation of extracellular membrane and progression of tumor cells, and PAI-1 gene polymorphic variants have been known as the prognostic biomarkers of non-small cell lung cancer patients. Recently, experimental in vitro study revealed that transforming growth factor-${\beta}1$ initiated PAI-1 transcription through epithelial growth factor receptor (EGFR) signaling pathway. However, there is little clinical evidence on the association between PAI-1 A15T gene polymorphism and prognosis of Korean population with pulmonary adenocarcinoma and the influence of activating mutation of EGFR kinase domain. Methods: We retrospectively reviewed the medical records of 171 patients who were diagnosed with pulmonary adenocarcinoma and undergone EGFR mutation analysis from 1995 through 2009. Results: In all patients with pulmonary adenocarcinoma, there was no significant association between PAI-1 A15T polymorphic variants and prognosis for overall survival. However, further subgroup analysis showed that the group with AG/AA genotype had a shorter 3-year survival time than the group with GG genotype in patients with EGFR mutant-type pulmonary adenocarcinoma (mean survival time, 24.9 months vs. 32.5 months, respectively; p=0.015). In multivariate analysis of 3-year survival for patients with pulmonary adenocarcinoma harboring mutant-type EGFR, the AG/AA genotype carriers had poorer prognosis than the GG genotype carriers (hazard ratio, 7.729; 95% confidence interval, 1.414-42.250; p=0.018). Conclusion: According to our study of Korean population with pulmonary adenocarcinoma, AG/AA genotype of PAI-1 A15T would be a significant predictor of poor short-term survival in patients with pulmonary adenocarcinoma harboring mutant-type EGFR.

Solid-Phase Refolding of Poly-Lysine fusion Protein of hEGF and Angiogenin (Poly-lysine이 연결된 hEGF와 angiogenin의 융합단백질의 고체상 재접힘)

  • Park, Sang-Joong;Ryu, Kang;Suh, Chang-Woo;Chai, Young-Gyu;Kwon, Oh-Byung;Park, Seung-Kook;Lee, Eun-Kyu
    • KSBB Journal
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    • v.17 no.2
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    • pp.153-157
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    • 2002
  • A fusion protein, consisting of a human epidermal growth factor as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as an inclusion body in recombinant E. coli, yet when the conventional solution-phase refolding process was used the refolding yield was very low due to severe aggregation, probably because of the opposite surface charge resulting from the vastly different pl values of each domain. Accordingly the solid-phase refolding process, which exploits the ionic interactions between a solid matrix and the protein, was tried, however the ionic binding yield was also very low regardless of the resins and pH conditions used. Therefore, to provide a higher affinity toward the solid matrix, six Iysine residues were tagged to the N-terminus of the hEGF domain. When cation exchange resins, such as heparin- or CM-Sepharose, were used as the matrix, the adsorption capacity increased 2.5~3-fold and the subsequent refolding yield increased nearly 15-fold compared to the conventional process. A similat result was also obtained when an Ni-NTA metal affinity resin was used.

In vitro Evaluation of Biological Activities of Wa-song (Orostachys japonicus A. Berger) and Korean Traditional Plants Mixture (와송과 한약재 복합물의 in vitro 생리활성 평가)

  • Lee, Soo-Jung;Shin, Jung-Hye;Kang, Jae-Ran;Hwang, Cho-Rong;Sung, Nak-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.3
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    • pp.295-301
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    • 2012
  • This study was carried out to determine the biological activities of Wa-song (Orostachys japonicus) hot water extracts. Four types of extract samples were prepared, including Wa-song, traditional plants mixture [PM; mixture of Baekbokryung (Poria cocos), Changchul (Atractylodis rhizoma), and Sa-in (Amomum xanthoides)], and two different ratio composites of these (mixture of PM and Wa-song extract, 1:1 (v/v); PMO-1 and 1:3 (v/v); PMO-3). Their biological activities were measured using various in vitro assays. Total phenolic and flavonoid contents of PM were higher compared to those of Wa-song, and those of PMO-1 were higher than those of PMO-3. Further, PMO-1 contained higher ABTS and DPPH radical scavenging, reducing power, and nitrite scavenging activities than PMO-3. On the contrary, PMO-3 contained higher tyrosinase and inhibitory activities of MCF-7 and HT-29 cancer cells than PMO-1. According to the results, biological activities of PMOs were significantly higher than those of Wa-song extract and PM in in vitro assays. Therefore, we expect that PMOs could show higher biological activities than Wa-song extract alone in vivo.

Effect of p-Phenylenediamine to Rat Skin (p-Phenylenediamine이 흰쥐의 피부 조직에 미치는 영향)

  • Lee, SangHee;Cho, HyunGug;Lee, Sang-Il
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.9
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    • pp.1330-1335
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    • 2005
  • Red brownish p-pheylenediamine (PPD) has been widely used hair dye for women. The dye was known to cause systemic anaphylaxis, dermatitis and bladder cancer. But the effect of PPD toxicity with oxygen free radical has not been studied. This study investigated the degree of skin injury by PPD. PPD ($2.5\%$ PPD in $2\%\;NH_{4}OH$) was applied to the rat skin ($25 mg/16.5\;cm^2$) 3 or 5 times every other day. Histopathological findings demonstrated the proliferation of epithelial cells and the increased keratinization by PPD. The activities of glucose 6-phosphatase (G6Pase) was decreased and acid phosphatase (ACP) was increased in PPD-applied rat skin. Groups in which PPD was applied 5 times were more damaged than groups applied 3 times. To examine the relationship between tissue damage and oxygen free radicals, effect of PPD on xanthine oxidase (XO) activity was measured and XO activity was more significantly increased in the group treated with PPD 5 times than 3 times. However, reduced glutathione (GSH) content, and the activities of catalase (CAT), super-oxide dismutase (SOD) and glutathione S -transferase (GST) were more decreased in PPD-applied groups than in controls. Even though the activities of XOD was not changed in the group treated with PPD 3 times, the decreased activities of oxygen free radical system and the damaged skin tissue were observed. This result might be caused by the production of toxic PPD metabolites in rat skin. In conclusion, topical PPD application led to skin injury in a dose-dependent manner, probably due to the generation rate of oxygen free radical.

Role of Blood Flow vs. $O_{2}$ Consumption in Nicotinamide-induced Increase $pO_{2}$ in a Murine Tumor (Nicotinamide에 의한 종양내 산소 분압의 증가에 있어서 혈류 또는 산소 소모의 역할)

  • Lee Intae;Demhartner Thomas J.;Cho Moon-June
    • Radiation Oncology Journal
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    • v.12 no.1
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    • pp.17-25
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    • 1994
  • We evaluated the effect of nicotinamide on cellular $O_{2}$ consumption and metabolic status i.e., adenylate phosphates and $NAD^{+}$in-vitro, and changes in blood flow in-vivo, to determine whether changes in cellular metabolism or increased oxygen availability, was responsible for increased tumor oxygenation. Thirty min, pre-incubation of cells with$\∼$4mM (= 500mg/kg) nicotinamide resulted in no change in cellular $O_{2}$ consumption. Similarly neither the adenylate Phosphates nor the cellular $NAD^{+}$levels were altered in the presence of $\∼$4mM nicontinamide. In-vivo, nicotinamide (500mg/kg) increased $O_{2}$ availability as estimated by changes in relative tumor blood flow (RBC flux). The changes in RBC flux measured by the laser Doppler method, were tumor volume dependent and increased from$\∼$35$ \% $ in$\∼$ 150$mm_{3}$tumors to$\∼$~75$ \% $ in$\∼$500$mm^{3}$ tumors. In conclusion, these observations indicate a reduction in local tissue $O_{2}$ consumption is not a mechanism of improved tumor oxygenation by nicotinamide in FSa II murine tumor model. The primary mechanism of increased $pO_{2}$ appears to be an increased local tumor blood flow.

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Effects of Saururus Chinensis Baill Extracts on Anticancer Activity and Cadmium Induced Cytotoxicity (삼백초 추출물의 항암활성과 카드뮴에 대한 독성억제효과)

  • Park Seung-Joung;Yoo Hyun-Ju;Choi Hee-Seok;Seo Byeong-Yun;Yang Seon-Ho;Kim Yeong-Hun;Jeong Jae-Yeal;Lee Ki-Nam
    • Journal of Society of Preventive Korean Medicine
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    • v.8 no.2
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    • pp.81-98
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    • 2004
  • 1. The cell viability was determined by MTT method. Their cytotoxic activities against three cancer cell lines such as A549, MDA-MB-231 and SNU-C4 cell line were tested. Among them, The methanol extract of Saururus chinensis Baill. showed the strongest cytotoxic effect against SNU-C4 cells. These results suggest that the methanol extract of Saururus Chinensis Baill. possessed a potential antitumorous agent. 2. In vitro the antitoxic activity of ethanol extract of Saururus Chinensis Bail on NIH 3T3 fibroblasts was evaluate by the MTT (3-(4,5-dimethyl- thiazol-2-yl)-2,5-diphenyl-2H-tetra캐lium bromide) and SRB (sulforhodamine B protein) assays. The number of NIH 3T3 fibroblasts were increased and tend to regenerate. These results suggest that Saururus Chinensis Bail extract retains a potential antitoxic activity. 3. Complexation of Cd (II) ion with ligands such as quercetin has been determined by UV-vis spectrophotometric method in tris buffer solution at various pH. It was found that only 1 : 1 Cd-complex is formed by the interaction between catechol moiety in ring B and cadmium [Cd(II)] in aqueous solution. The spectral parameters for Cd-complex were determined by Beer's law at various pH. It has shown that 1 : 1 Cd-complex has a maximum absorbance and red shift by the alkaline pH.

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Establishment of I-131, Tc-99m Labeling Methods to In-house Anti-CEA Antibodies and Evaluation of the Immunological Characteristics (국산 항 CEA 항체의 I-131, Tc-99m 표지법 확립 및 면역학적 특성 분석)

  • Chung, June-Key;Lee, Dong-Soo;Lee, Myung-Chul;Chung, Hong-Keun;Koh, Chang-Soon;Hong, Mee-Kyoung;Choi, Seok-Rye;Seo, Il-Taek;Chung, Jun-Ho
    • The Korean Journal of Nuclear Medicine
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    • v.26 no.2
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    • pp.346-354
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    • 1992
  • Cancer cells have several tumor-associated antigens on the cell surfaces, and antibodies against these antigens have been developed by many investigators. Radiolabeled antibodies have been used as new methods to diagnose and treat malignant tumors. Especially anti-carcinoembryonic antigen (CEA) is the most popular antibody for these purposes. In this investigation, we tried to label $^{131}I$ and $^{99m}Tc $ to anti-CEA monoclonal antibodies which were developed in the Seoul National University College of Medicine. We found CEA-79 and CEA-92 antibodies had the better immunological characteristics among 8 anti-CEA monoclonal antibodies. And radioiodination of CEA-79 could be performed by chloramine-T method, while radioiodination of CEA-92 by iodogen method. To label these antibodies with $^{99m}Tc $, we used pretargeting transchelation as direct labeling method. At first, $^{99m}Tc $ was bound to glucaric acid, and monoclonal antibody was reduced by $\beta-mercaptoethanol$. When these were incubated together. $^{99m}Tc $ bound to glucarate was switched to monoclonal antibody because of higher affinity. We established conditions of several steps in this method. Anti-CEA monoclonal antibodies labeled with $^{131}I$ and $^{99m}Tc $ are expected to be used valuably in the detection and treatment of malignant tumors.

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Beneficial Effect of Curcumin on Epidermal Permeability Barrier Function in Hairless Rat (무모쥐에서 자외선에 의한 피부 장벽 손상에 미치는 커큐민의 보호 효과)

  • Jeon, Hee-Young;Kim, Jeong-Kee;Kim, Wan-Gi;Lee, Sang-Jun
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.686-690
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    • 2008
  • Recent research has shown that curcumin has beneficial effects in a variety of skin diseases, including scleroderma, psoriasis, and skin cancer. In this study, we assessed the effects of curcumin on epidermal permeability barrier function in vivo and in vitro. In order to evaluate the effects of curcumin on epidermal permeability barrier function in vivo, hairless rats were exposed to UVB irradiation, and curcumin was administered orally at a dosage of 150 mg/kg per day for 8 weeks. Transepidermal water loss (TEWL) and epidermal thickness were measured at the end of the experiment. The expression of filaggrin, a marker of keratinocyte differentiation, and serine palmitoyltransferase (SPT), a marker of the formation of the stratum corneum lipid barrier, in human HaCat keratinocytes were analyzed. The in vivo results showed that an 8 week administration of curcumin markedly prevented the UVB-induced increase in TEWL. The UV-induced increase in epidermal thickness was also reduced significantly by curcumin treatment. The in vitro results demonstrated the concentration-dependent effects of curcumin on the expression of both filaggrin and SPT in HaCat cells, reflecting the notion that curcumin can induce epidermal keratinocyte differentiation and can improve the recovery of skin barrier functions. These results show that curcumin is a promising candidate for the improvement of epidermal permeability barrier function.

Clinical and Immunohistochemical Characteristics of Thyroid Papillary Cancer with Lateral Neck Lymph Node Metastasis (측경부 림프절 전이를 동반한 갑상선 유두상암의 임상 양상과 면역조직화학적 특성)

  • Lee, Seon-Uk;Jin, Sung-Min;Lee, Sang-Hyuk;Sohn, Jin-Hee;Chae, Seung-Wan;Kim, Dong-Hoon
    • Korean Journal of Head & Neck Oncology
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    • v.27 no.1
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    • pp.59-65
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    • 2011
  • Background and Objectives : Papillary thyroid carcinoma(PTC) frequently metastasize to the regional neck, however, lateral neck lymph node metastasis is less common. The aim of this study is to investigate clinical and immunohistochemical features of PTC with lateral LN metastasis, and determine the predictive factors for lateral LN metastases. Material and Methods : We undertook a retrospective study of 83 patients treated between January 2007 and December 2009 for PTC by thyroidectomy with or without lateral neck dissection. The following criteria were used to study the clinical predictive value of lateral LN. metastases : sex, age, tumor size, multifocality, extracapsular spread(ECS) and lymphovascular emboli. Immunohistochemical staining for VEGF-A, VEGF-C, Bax, Bcl-2, Cyclin D1, Cyclin E, $p27^{kip1}$ and $p57^{kip2}$ was performed, and quantified blindly by three pathologists who had no clinical information of the patients. Immunohistochemical expression was scored as high(>50% of cells stained) or low(0-49%). Results : With use of univariate and multivariate analysis, tumor size(>2cm) and ECS were independent correlates of lateral LN metastasis in PTC. Expression of VEGF-C, Bax, and Cyclin D1 in the PTC with lateral LN metastasis was scored higher than in PTC without lateral LN metastasis(p<0.05). Conclusion : The important risk factors for lateral LN metastasis in PTC are primary tumor size and the presence of ECS. And expression of VEGF-C, Bax and cyclin D1 may be considered of lateral LN metastatic potential in PTC.

Cytotoxic Flavonoids from the Whole Plants of Chrysanthemum zawadskii Herbich var. latilobum Kitamura (구절초에서 분리한 Flavonoids의 인체암 세포주에 대한 세포독성 효과)

  • Kwon, Hyun-Sook;Ha, Tae-Joung;Hwang, Seon-Woo;Jin, Young-Min;Nam, Sang-Hae;Park, Ki-Hun;Yang, Min-Suk
    • Journal of Life Science
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    • v.16 no.5
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    • pp.746-749
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    • 2006
  • Two known flavonoids, luteolin (1) and acacetin (2) were isolated from a $CHCI_3$ soluble fraction of the whole plants of Chrysanthemum zawadskii Herbich var. latilobum Kitamura, and their structures were determined by NMR analysis. The luteolin (1) was isolated from this plant for the first time. These compounds were examined for their in vitro cytotoxic activities against four human cancer cell lines including HCT116 (colon), UO-31 (renal), PC-3 (prostate) and A549 (lung) by sulforhodamine B(SRB) assay. Acacetin (2) showed significant cytotoxic activity against HCT116 and UO-31 cells with an $IC_{50}$ of 2.44 and $2.89\;{\mu}g/ml$, respectively.