• Journal of Yeungnam Medical Science
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• v.3 no.1
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• pp.103-110
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• 1986

Cytochrome P-450(CP-450) is one of the three components of the liver microsomal enzyme system which hydroxylates fatty acids, hydrocarbons and a variety of drugs and other foreign compounds. Female Balb/c mice were immunized with purified polychlorinated bipheny(PCB)-induced CP-450 LMII. The spleen cells derived from immunized mice were fused with $SP^2$ myeloma cells using polyethylene glycol(PEG 3500). The hybrid cells were selected by hypoxanthine-aminopterine and thymidine(HAT) medium and the culture fluid were screened by enzyme-linked immunosorbent assay to CP450 LMII. The hybrid cess(${\times}10^7$) were innoculated into intraperitoneal cavity of Balb/c mice for the purpose of production of ascitic fluids. Monoclonal antibody(Mab) was purified from ascitic fluid by DEAE cellulose ion exchange chromatography and $I^{125}$-labeled Mab was also confirmed by autoradiography and SDS-polyacrylamide gel electrophoresis (MW : 55,000 and 110,000).

• Journal of Yeungnam Medical Science
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• v.34 no.1
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• pp.43-54
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• 2017

Background: Extracellular sulfatases (Sulfs), sulfatase 1 (Sulf1) and sulfatase 2 (Sulf2), play a pivotal role in cell signaling by remodeling the 6-O-sulfation of heparan sulfate proteoglycans on the cell surface. The present study examined the effects of Sulfs on angiotensin II (Ang II)-induced hypertensive mediator expression and vascular smooth muscle cells (VSMCs) proliferation in spontaneously hypertensive rats (SHR). Methods: Ang II receptors, 12-lipoxygenase (12-LO), and endothelin-1 (ET-1) messenger RNA (mRNA) expressions in SHR VSMCs were analyzed by real-time polymerase chain reaction and Western blotting. VSMCs proliferation was determined by [$^3H$]-thymidine incorporation. Results: Basal Sulfs mRNAs expression and enzyme activity were elevated in SHR VSMCs. However, Sulfs had no effect on the basal or Ang II-induced 12-LO and ET-1 mRNA expression in SHR VSMCs. The inhibition of Ang II-induced 12-LO and ET-1 expression by blockade of the Ang II type 2 receptor ($AT_2\;R$) pathway was not observed in Sulf1 siRNA-transfected SHR VSMCs. However, Sulf2 did not affect the action of $AT_2\;R$ inhibitor on Ang II-induced 12-LO and ET-1 expression in SHR VSMCs. The down-regulation of Sulf1 induced a reduction of $AT_2\;R$ mRNA expression in SHR VSMCs. In addition, the inhibition of Ang II-induced VSMCs proliferation by blockade of the $AT_2\;R$ pathway was mediated by Sulf1 in SHR VSMCs. Conclusion: These findings suggest that extracellular sulfatase Sulf1 plays a modulatory role in the $AT_2\;R$ pathway that leads to an Ang II-induced hypertensive effects in SHR VSMCs.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.12
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• pp.4995-5000
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• 2014

Background: Cancer constitutes a key pressure on public health regardless of the economy state in different countries. As a kind of highly malignant epithelial tumor, lacrimal gland adenoid cystic carcinoma can occur in any part of the body, such as salivary gland, submandibular gland, trachea, lung, breast, skin and lacrimal gland. Chemotherapy is one of the key treatment techniques, but drug resistance, especially MDR, seriously blunts its effects. As an element of the 60S large ribosomal subunit, the ribosomal protein L39-L gene appears to be documented specifically in the human testis and many human cancer samples of different origins. Materials and Methods: Total RNA of cultured drug-resistant and susceptible lacrimal gland adenoid cystic carcinoma cells was seperated, and real time quantitative RT-PCR were used to reveal transcription differences between amycin resistant and susceptible strains of lacrimal gland adenoid cystic carcinoma cells. Viability assays were used to present the amycin resistance difference in a RPL39-L transfected lacrimal gland adenoid cystic carcinoma cell line as compared to control vector and null-transfected lacrimal gland adenoid cystic carcinoma cell lines. Results: The ribosomal protein L39-L transcription level was 6.5-fold higher in the drug-resistant human lacrimal gland adenoid cystic carcinoma cell line than in the susceptible cell line by quantitative RT-PCR analysis. The ribosomal protein L39-L transfected cells revealed enhanced drug resistance compared to plasmid vector-transfected or null-transfected cells as determined by methyl tritiated thymidine (3H-TdR) incorporation. Conclusions: The ribosomal protein L39-L gene could possibly have influence on the drug resistance mechanism of lacrimal gland adenoid cystic carcinoma cells.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.1070-1076
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• 2009

Telomerase reverse transcriptase (TERT), which prolongs the replicative life span of cells, is highly upregulated in 85-90% of human cancers, whereas most normal somatic tissues in humans express limited levels of the telomerase activity. Therefore, TERT has been a potential target for anticancer therapy. Recently, we described a new approach to human cancer gene therapy, which is based on the group I intron of Tetrahymena thermophila. This ribozyme can specifically mediate RNA replacement of human TERT (hTERT) transcript with a new transcript harboring anticancer activity through a trans-splicing reaction, resulting in selective regression of hTERT-positive cancer cells. However, to validate the therapeutic potential of the ribozyme in animal models, ribozymes targeting inherent transcripts of the animal should be developed. In this study, we developed a Tetrahymena-based trans-splicing ribozyme that can specifically target and replace the mouse TERT (mTERT) RNA. This ribozyme can trigger transgene activity not only also in mTERT-expressing cells but hTERT-positive cancer cells. Importantly, the ribozyme could selectively induce activity of the suicide gene, a herpes simplex virus thymidine kinase gene, in cancer cells expressing the TERT RNA and thereby specifically hamper the survival of these cells when treated with ganciclovir. The mTERT-targeting ribozyme will be useful for evaluation of the RNA replacement approach as a cancer gene therapeutic tool in the mouse model with syngeneic tumors.

• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.352-373
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• 2003

Rhamnose-rich (RROP-s) and fucose-rich (FROP-s) oligo-and polysaccharides were prepared and extensively characterised by physical and chemical procedures [1,2] and compared to L-fucose. Their biological properties were then studied on human skin fibroblast cell cultures, human skin explant cultures and on hairless rat skin, using a variety of cell-biological, biochemical and computerised morphometrical procedures. Among the most important properties we could establish, the following are of particular interest for the tretment and prevention of age-dependent modifications of human skin (loss of skin-tissue, cells and matrix, wrinkle formation and others) : stimulation of cell proliferation (by $^3$[H]-thymidine incorporation and the MTT test), scavenging of reactive oxygen species (ROS) using several different procedures, and protease (MMP-2 and MMP-9) down-regulation. A topical preparation, using RROP-s and FROP-s, and/or L-fucose, was shown to increase cell proliferation, dermal matrix synthesis, efficient scavenging of ROS-s and to increase also the thickness of dermal tissue when applied for 4 weeks on hairless rat skin, accompanied by the densification of collagen bundles as well as by an increase of elastin synthesis. Using fluorescent labeled FROPs, it could be shown that these oligosaccharides react with cell-membrane receptors and especially with the elastin-laminin-receptor and the fucose-mannose receptor, but they penetrate also in the cell nucleus, suggesting the possibility of a direct action on the regulation of gene expression. When applied to the human skin of a team of voluntary women encompassing all age-groups, the efficiency of FROP-containing preparation could be confirmed using indentometry and computerised evaluation of skin micro-relief, as well as evaluation of periorbital wrinkles. It appears therefore that these preparations correspond to all the requirements of active anti-aging principles.

• Nuclear Engineering and Technology
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• v.38 no.3
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• pp.285-292
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• 2006

Thermoresistant (TR) clones of radiation-induced fibrosarcoma (RIF) cells have been reported to show an adaptive response to 1cGy of low dose radiation, and HSP25 and inducible HSP70 are involved in this process. In this study, to further elucidate the mechanism by which HSP25 and inducible HSP70 regulate the adaptive response, HSP25 or inducible HSP70 overexpressed RIF cells were irradiated with 1cGy and the cell cycle was analyzed. HSP25 or inducible HSP70 overexpressed cells together with TR cells showed increased G1 phase after 1cGy irradiation, while RIF cells did not. $[^3H]-Thymidine$ and BrdU incorporation also indicated that both HSP25 and inducible HSP70 are involved in G1 arrest after 1cGy irradiation. Molecular analysis revealed upregulation of p27Cip/Kip protein in HSP25 and inducible HSP70 overexpressed cells, and cotransfection of p27Cip/Kip antisense abolished the induction of the adaptive response and 1cGy-mediated G1 arrest. The above results indicate that induction of an adaptive response by HSP25 and inducible HSP70 is mediated by upregulation of p27Cip/Kip protein, resulting in low dose radiation-induced G1 arrest.

• YAKHAK HOEJI
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• v.38 no.6
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• pp.806-813
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• 1994

These experiments were conducted to investigate the effects of Cervi cornu extract on lymphocyte blastogenesis in spleen, thymus, lymph node, born marrow cells of Balb/c mouse, haemagglutination reaction against sheep red blood cell (SRBC), plaque forming cell (PFC) assay against SRBC and IL-2 production. Lymphocyte blastogenesis was determined by $[^3H]-thymidine$ incorporation. According to the lymphcoyte blastogenesis test on the immune cell. Ceriv cornu extrat was showed a potent mitogenic activity on the spleen and lymph node cells, but had mild mitogenic activity on the thymus and born marrow cells. Mitogenic active component of Crevi cornu extract was identified to be materials where molecular weights are higher than 5,000 by membrane filteration method. Cervi cornu extrat was shown to increase mitogenic effect on the lipopolysaccharide (LPS)-stimulated spleen cells significantly, but decrease mitogenic effect on the Con A stimulated spleen cell at the concentration 0.3%, 1% and 3%. Ceriv cornu extract didn't show to be haemagglutination reaction and showed to inhibit the Con A-induced haemagglutination reaction against SREC. Result of SRBC-PEC test. Ceriv cornu extract significantly increase the number of PEC at the concentration of 0.1% and 1%. When IL-2 or IL-4 production was determined by proliferation of CTLL-2 cells. Ceriv cornu extract was not shown to stimulate the production of IL-2. From the above results, it is shown that Ceriv cornu extract increased antibody production by B cells, but nor IL-2 production by helper T cells.

• The korean journal of orthodontics
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• v.21 no.1 s.33
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• pp.97-111
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• 1991

It is the aim of this study to investigate the effects of sodium fluoride and sodium orthovanadate upon the proliferation and activity of the osteoblast (MC3T3-E1 cells). MC3T3-E1 cells were cultured in $\alpha-MEM$ containing $10\%$ FBS and various concentration of sodium fluoride and sodium orthovanadate was appended to serum free media. DNA synthesis was examined through the $[^3H]$ thymidine incorporation into DNA. Collagen synthesis was examined through the $[^3H]$ proline incorporation into collagenase digestible protein and noncollagen protein. The following results were drawn; 1. Sodium fluoride stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M$ to $10{\mu}M$ (P < 0.005). 2. Sodium orthovanadate stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M\;to\;8{\mu}M$, however showed diminution at $10{\mu}M$ (P < 0.001). 3. Sodium fluoride and sodium orthovanadate stimulated the percent collagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M$ to $10{\mu}M$ (P < 0.001). 4. Sodium fluoride and sodium orthovanadate stimulated the noncollagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M\;to\;10{\mu}M$ (P < 0.001). In conclusion, sodium fluoride and sodium orthovanadate stimulate the proliferation and activity of osteoblast by stimulation of DNA synthesis and collagen and noncollagen synthesis in osteoblast.

• Journal of the East Asian Society of Dietary Life
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• v.4 no.3
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• pp.59-65
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• 1994

Growth-stimulating effects of cow's milk was examined using Vero cell cultre. Medium containing whole cow's milk stimulated cell growth about the same degree as that containing fetal bovine serum. The growth-stimulating factor in cow's milk was purified using hydrophobic (phenyl-sepharose) and gel filtration (Sephadex G-100) column chromatographies. It appeared that the factor is a highly hydrophobic protein, since the major growth-stimulating activity was found in the fractions eluted with 50% ethylene glycol from the phenyl-sepharose column during the purification. The purified factor showed a single band on the polyacrylamide gel electrophoresis in the presence of 1% (w/v) SDS. The factor has been found to have a relatively high molecular weight in the range of about Mr=100,000-150,000. In the presence of the purified factor (5%, w/v) in the culture medium, the incorporation of [3H]-thymidine into the cells was increased approximately 2,400-fold over that in the presence of 5% (w/v) fetal bovine serum. It seems that the growth-stimulating factor purified in this study is one of the major growth factors in the cow's milk.

• The Journal of Internal Korean Medicine
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• v.12 no.2
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• pp.1-15
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• 1991

Bigihwan which has been widely used in Oh-jug in oriental Medicine was investigated on its antitumor effect employing blood cancer cell lines. K 562 derived from human erytholeukemia, Raji from lymphoma and $MO_4$ from hlastogenic cancer were used in this study to see the analytical evaluation of Bigihwan' s antitumor effect using three different kinds of methods such as $^{3}H-thymidine$ up take assay. MTT assay and live cell counts by Trypan blue assay. The result obtained are as follows. 1. When higher than 10% Bigihwan was treated. inhibitory effect of tumor killing action was observed showing the increasing order of $MO_4$, K 562 and Raji(Fig. 3). 2. When 1 to 5% of Bigi-hwan was treated, 4 to 30% of tumor cell survival was observed according to various blood tumor cell lines suggesting that antitumor effect of Bigi-hwan was different as the characteristics of tumor cells showing 70 to 95% cell killing effent(Fig. 4). 3. Compared the survivals of cells by relative scales though the initial cpm was variable because of different cell growth rate. Raji was most effective being killed 95% by the treatment of 1% Bigihwan while Raji and K562 showed 93% by 5% Bigihwan.(Fig. 5) 4. The survival rate of Raji derived from Burkitt lymphoma was rather increased to 2.3 times when Bigihwan concentration was increased from 1 to 10% lmplying of refraining from over use of this anticancer drug. specially to lymphoma patients(Fig. 5). 5. Bigihwan was most effective to K 562 and then $MO_4$ showing 95% tumor cell death by using 1% of this anticancer drug while it was least effective to Raji showing only 68% of tumor cell death(Fig.7). 6. Judging from the all the analytical methods used in this study, through all different three tumor cell lines. Bigihwan was most effective to K 562 derived from human erythroleukemia.