• Asian Pacific Journal of Cancer Prevention
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• v.13 no.4
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• pp.1097-1104
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• 2012

Background/Aim: Pristimerin isolated from Celastrus and Maytenus spp can inhibit proteasome activity. However, whether pristimerin can modulate cancer metastasis is unknown. Methods: The impacts of pristimerin on the purified and intracellular chymotrypsin proteasomal activity, the levels of regulator of G protein signaling 4 (RGS 4) expression and breast cancer cell lamellipodia formation, and the migration and invasion were determined by enzymatic, Western blot, immunofluorescent, and transwell assays, respectively. Results: We found that pristimerin inhibited human chymotrypsin proteasomal activity in MDA-MB-231 cells in a dose-dependent manner. Pristimerin also inhibited breast cancer cell lamellipodia formation, migration, and invasion in vitro by up-regulating RGS4 expression. Thus, knockdown of RGS4 attenuated pristimerin-mediated inhibition of breast cancer cell migration and invasion. Furthermore, pristimerin inhibited growth and invasion of implanted breast tumors in mice. Conclusion: Pristmerin inhibits proteasomal activity and increases the levels of RGS4, inhibiting the migration and invasion of breast cancer cells.

• Journal of Chest Surgery
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• v.26 no.2
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• pp.80-85
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• 1993

Synthetic and biosynthetic vascular grafts of small diameter have long been considered to be prone to thrombosis, ultimately leading to the complete graft occlusion. Endothelial cell seeding onto synthetic blood-contacting surfaces has been suggested to be an ideal means to solve this problem. This study described a culture method of bovine endothelial cells and evaluated blood-compatibility and seeding efficiency of cultured endothelial cells. Bovine pulmonary artery endothelial cells were harvested enzymatically and grown to confluence on polystyrene culture flask surfaces using established techniques. The identification of endothelial cells was made through the demonstration of expression of factor VIII R:Ag by immunofluorescent technique. To quantitate the effect of improvement in blood-compatibility of viable endothelial cells, endothelial monolayers were exposed to blood containing $\^$111/In-oxine labeled platelets. Viable endothelial monolayers retained less labeled platelets than control surfaces. The Indium-labeled endothelial cells were seeded onto three different blood-contacting surfaces of Dacron vascular graft immobilized in specially equipped wells and incubated for specific time intervals (t=15, 30, 60, 120 minutes). Longer incubation times showed improved cell adherence in collagen-coated and fibrin-coated Dacron vascular graft groups. However in untreated Dacron grafts, no direct relationship was observed between incubation time and endothelial cell seeding efficiency. This may be due to leakage of endothelial cells through porosity of Dacron grafts in this in-vitro experimental condition.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.494-498
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• 1989

Hybridoma cell lines, which secrete monoclonal antibodies (mAbs) against the surface antigens of Cryptosporidium parvum Sporozoites, were produced by fusing spleen cells of C. parvum Sporozoite-immunized mice with P3-X63-Ag8 myeloma cells. Two cloned antibody-secreting cell lines, Kor1 and Ea2, were established and produced IgG1 and IgG2a antibodies, respectively. Percoll-purified sporozoites were solubilized and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Western blot assay demonstrates that an antigen of 20-kDa was bound by monoclonals. By indirect immunofluorescence microscopy, mAb exhibited uniform binding to the sporozoite surface.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.35 no.2
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• pp.55-65
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• 2009

Purpose: We determined the therapeutic effects of blockade of epidermal growth factor(EGF) and vascular endothelial growth factor(VEGF) receptor tyrosine kinases on the growth of oral squamous cell carcinoma(OSCC) xenografted in athymic nude mice. Experimental Design: We investigated the in vivo antitumor effects of a tyrosine kinase inhibitor for EGFR and VEGFR-2, AEE788 in a mouth floor(orthotopic) tumor model. Nude mice with orthotopic tumors were randomized to receive AEE788, paclitaxel, a combination of AEE788 and paclitaxel, or control. Antitumor mechanisms of AEE788 were determined by immunohistochemical/immunofluorescent and apoptosis assays. Results: Tumors of mice treated with AEE788 demonstrated down-regulation of phosphorylated EGFR, phosphorylated VEGFR and their downstream mediators(pMAPK and pAkt), decreased proliferative index, decreased microvessel density(MVD). As a result, growth of the primary tumor and nodal metastatic potentials were inhibited by AEE788. Conclusion: These data show that EGFR and VEGFR can be molecular targets for the treatment of OSCC.

• The Korean Journal of Nuclear Medicine
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• v.25 no.1
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• pp.110-116
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• 1991

To elucidate alteration of peripheral T cell subsets in thyroid tumors, the author enumerated T cell subsets in periphral blood by indirect immunofluorescent method, using monoclonal antibodies (CD3, CD4 and CD8) in 17 cases of thyroid cancer, 12 cases of thyroid adenoma, and 16 cases of adult healthy subjects as controls. Diagnoses were confirmed histopatologically in thyroid cancer and adenoma, and were established on the basis of commonly accepted clinical and biochemical criteria in Hashimoto's thyroiditis. The blood was drawn from veins of the patients and control subjects in Pusan National University Hospital during the period of January to October 1990. The results obtained were summarized as follow: 1) The percentage of CD3+ cells was significantly decreased in thyroid cancer as compared with healthy subjects. 2) The percentage of CD4+ cells was not different among thyroid cancer, thyroid adenoma, Hashimoto's thyroiditis and control subjects each other. 3) The percentage of CD8+ cells was significantly decreased in thyroid cancer as compared with adult healthy subjects, and tended to be decreased as compared with thyroid adenoma and Ha-shimoto's thyroiditis. 4) The CD/CD8 ratio was significantly increased in thyroid cancer as compared with control subjects, and tended to be increased as compared with thyroid adenoma and Hashimoto's thyroiditis. On the basis of the results, it can be suggested that the immunodysfunction may be due to decreased soppressor/cytotoxic T cells in thyroid cancer.

• Korean Journal of Microbiology
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• v.33 no.2
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• pp.97-104
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• 1997

Herpes simpex virus type 2 (HSV-2) infects the genital and oral mucosae of human and other animals. HSV-2 infection is a widespread health problem causing various clinical syndromes including oral, genital, and ocular lesions, viral encephalitis, and recurrent diseases. Hybridorna cell lines secreting a monoclonal antibody (mAb) against the HSV-2 were produced by fusing spleen cells of HSV-2-immunized mice with Sp2/0-AgI4 myeloma cells. One hybridoma cell line was established and its monoclonal C-2, IgM, recognized the antigens of 134, 86, and 43 kDa in western blot analysis. In SDS-P AGE analysis of HSV -2 antigens, 25 bands were separated between 3D kDa and 159 kDa. In indirect immunofluorescent assay, mAbs exhibited binding to the virus antigen expressed on Vero cell infected with HSV-2.

• Archives of Plastic Surgery
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• v.33 no.6
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• pp.757-760
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• 2006

Purpose: Anaplastic large cell lymphoma, has the following three characteristics of a malignant lymphoma; 1) An irregular large nucleus, called pathologic atypical cells, 2) Eosinophilic cytoplasm, 3) Immunologically positive for Ki-1. Anaplastic large cell lymphoma occurs mostly in the lymph nodes, but about 40% has been observed to occur in other tissues. Skin is the one of the main sources of origin and it is called 'primary cutaneous anaplastic large cell lymphoma'. Methods: A 69-year-old male patient with an erythematous nodule, sized $1.5{\times}1.7cm$ on his right hand dorsum was excised under local anesthesia and on biopsy was diagnosed as 'Dermatofibrosarcoma Protuberans'. Three months after the local excision and biopsy, same natured mass reoccurred in the same region, and then spontaneous regressed after three weeks. However, metastatic large mass of $4.0{\times}5.0cm$, of same nature was observed on the elbow. The large mass was operated with wide excision and biopsy. Results: On final diagnosis, with an immunofluorescent stain with CD30(Ki-1), 'Primary cutaneous large cell lymphoma' was made. After follow up for three years, we did not observed recurrence and metastasis. Conclusion: We have reported that we have diagnosed primary cutaneous large cell lymphoma and treated without recurrence and metastasis.

• Biomedical Science Letters
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• v.13 no.4
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• pp.273-278
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• 2007

Stem cells have been the subject of increasing scientific interest because of their utility in numerous biomedical applications. Stem cells are capable of renewing themselves; that is, they can be continuously cultured in an undifferentiated state, giving rise to more specialized cells of the human body. Therefore, stem cells are an important new tools for developing unique, in vitro model systems to test drugs and chemicals and a potential to predict or anticipate toxicity in humans. In the present study, in vitro cultured F3 immortalized human neural stem cell line and in vivo adult Sprague Dawley rats was used to evaluate the cytotoxicity of anticancer drug paclitaxel. In vitro apoptotic activity of paclitaxel was evaluated in F3 cell line by a MTT assay and DAPI test. The cell death was induced with the treatment of 20 nM paclitaxel and chromatin degradation was detected by DAPI staining, which was analyzed by fluorescent microscope. In vivo studies, we also observed nestin immunoreactivity on subventricular zone, which is stem cell rich region in the adult brain of the SD rat. Immunofluorescent staining result shows that pixel intensities of nestin were decreased in a dose dependent manner. These results suggest that paclitaxel is able to induce cytotoxic activity both in F3 neural stem cell line and neural stem cell in SD rat brain.

• Korean Journal of Veterinary Research
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• v.29 no.3
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• pp.343-359
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• 1989

To study the effects of ibaraki virus on preimplantation mouse embryos collected from prepubertal ICR and BALB/cByJ mice (30~40days old) by superovulation, zona pellucidaintact(ZPI) or free(ZPF) embryos(n=774) of 4- to 8-cell and morulae were exposed to $10^{5.8}$ $TCID_{50}$ of the virus up to 96 hours. The embryos were examined morphologically by observing the degeneration and hatching rates, and virologically and immunologically by determining the presence of infection with the virus, in addition, the effect of washing the embryos to remove virus possibly attached to was also investigated. The ZPI 4- to 8-cell embryos and morulae exposed to the virus showed considerably higher degeneration rate than those not exposed, for 96, and for 72 to 96 hours, respectively(p<0.01). The ZPF 4- to 8-cell embryos and morulae exposed to the virus showed considerably higher degeneration rates than those not exposed, throughout the whole culture hours in vitro (p<0.01). The ZPI 4- to 8-cell embryos and morulae not exposed to the virus showed considerably higher rates of hatched blastocyst than those exposed (p<0.01). The virus infection rates of the ZPF 4- to 8-cell embryos and morulae were significantly higher than those of the ZPI embryos according to cell culture system. The viral antigen was detected exclusively on the zona pellucida of ZPI embryos, while the antigen was evenly distributed in the blastomeres of ZPF embryos by the immunofluorescent assay. In the ZPI embryos exposed to ibaraki virus, the virus was detected in the two times-washing groups, but not in the ten times-washing groups. The results indicated that zona pellucida of murine embryos would provide an effective protection and that ten times-washing of the ZPI embryos previously exposed to the virus was effective to remove virus from the embryos.

• BMB Reports
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• v.46 no.2
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• pp.65-72
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• 2013

In situ detection of RNAs is becoming increasingly important for analysis of gene expression within and between intact cells in tissues. International genomics efforts are now cataloging patterns of RNA transcription that play roles in cell function, differentiation, and disease formation, and they are demon-strating the importance of coding and noncoding RNA transcripts in these processes. However, these techniques typically provide ensemble averages of transcription across many cells. In situ hybridization-based analysis methods complement these studies by providing information about how expression levels change between cells within normal and diseased tissues, and they provide information about the localization of transcripts within cells, which is important in understanding mechanisms of gene regulation. Multi-color, single-molecule fluorescence in situ hybridization (smFISH) is particularly useful since it enables analysis of several different transcripts simultaneously. Combining smFISH with immunofluorescent protein detection provides additional information about the association between transcription level, cellular localization, and protein expression in individual cells.