• The Korean Journal of Physiology and Pharmacology
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• v.3 no.6
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• pp.597-603
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• 1999

Caldesmon (CaD), one of microfilament-associated proteins, plays a key role in microfilament assembly in mitosis. We have investigated the effects of overexpression of the high molecular weight isoform of CaD (h-CaD) on the physiology of vascular smooth muscle cells (VSMCs). Rat aortic VSMCs were stably transfected with plasmids carrying a full length human h-CaD cDNA under control of cytomegalovirus promoter. The majority of the overexpressed h-CaD appears to be localized predominantly on cytoskeleton structures as determined by detergent lysis. The overexpression of h-CaD, however, does not decrease the level of endogenous low molecular weight isoform of CaD. h-CaD overexpressing VSMCs (h-CaD/VSMCs) show a decreased growth rate than that of vector-only transfected cells when determined by $[^3H]thymidine$ uptake and cell counting after fetal bovine serum (FBS) stimulation. h-CaD/VSMCs were smaller than vector-transfected cells by 18% in cell diameter. These data suggest that overexpression of h-CaD can inhibit the poliferation and the cell volume of VSMCs stimulated by growth factors and that the gene therapy with h-CaD may be helpful to prevent the conditions associated with hypertrophy and/or hyperplasia of VSMCs after arterial injuries.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.3
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• pp.13-24
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• 2006

Purpose : 이 연구는 MCF-7 인간 유방암 세포주에 대한 활락효령단(活絡效靈丹) 추출물의 증식억제효과, 세포독성효과, 세포사 유발효과를 확인하기 위하여 이루어졌다. Methods : MCF-7 인간 유방암 세포주는 Dulbecco's modified Eagle's medium/F12 (DMEM/F12)에 10% fetal bovine serum (FBS)와 항생제를 가하여 만든 배지를 이용하여 배양하였고 MCF-7 세포를 96-well plate에 접종한 후 다양한 농도 (0 ${\sim}$ 2000 g/ml)의 활락효령단(活絡效靈丹)이 든 배지로 처리한 후 72시간 동안 배양하였고 또한 1000g/ml의 활락효령단(活絡效靈丹)이 든 배지로 처리한 후 48, 96, 192 시간동안 배양하여 각각 MTS assay kit로 세포생존율을 측정하였다. 세포독성은 Sulforhodamine B assay 방법을 이용해 측정하였고 세포사 과정에서 MCF-7 세포에서의 caspase 활성화를 측정하기 위해 Western blotting을 수행하여 poly ADP ribose polymerase (PARP)의 절단을 확인하였다. Results : 실험결과 활락효령단(活絡效靈丹) 추출물에 의한 세포성장 및 독성효과는 시간 및 농도에 비례하는 것으로 나타났고 세포고사과정에서 작용하는 caspase의 전기질인 PARP 절단량이 활락효령단(活絡效靈丹) 처리 농도에 비례에 증가하였다. Conclusions : 활락효령단(活絡效靈丹)은 다양한 기전에 의해서 유방암 세포에 대한 억제효과를 가질 수 있는 것으로 인식할 수 있다.

• Herbal Formula Science
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• v.14 no.2
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• pp.86-96
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• 2006

Objectives : This study wad designed to elucidate the short term effect of Rossa rugosae Radix on proliferation, differentiation and maturation of 3T3-L1 Preadipocyte. Methods : 3T3-L1 preadipocytes obtained from Korean Cell line Bank were cultured in a Dulbecco's modified eagle medium(DMED) culture colution containing 10% fetal bovine serum(FBS) and various concentration of aqueous extract of Rossa rugosae Radix on proliferation, differentiation and maturation of 3T3-L1 preadipocytes were investigate after treatment for 24 hours b measuring MTT, Oil Red O and latate dehydrogenase activity.. Results : The Rossa rugosae Radix extract inhibited significantly the proliferation of 3T3-L1 preadipocytes and tended to increase latate dehydrogenase activity in the media of differentiated 3T3-L1 preadipocytes & matured 3T3-L1 preadipocytes. the extract also inhibit the lipid accumulation of differentiated and maturaion of 3T3-L1 preadipocytes, suggesting that Rossa rugosae Radix has anti-obesity effect: however further in vivo study is needed to demonstrate its pharmacological effects.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.2
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• pp.358-364
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• 2006

To investigate the possible mechanism of Drynariae Rhizoma extracts as a candidate of anti-cancer drug, I examined the effects of Drynariae Rhizoma extracts on the apoptosis of U937 cell line. MTT assay, flow cytometric analysis, SDS-polyacrylamide gel electrophoresis, Western blot analysis, and RT-PCR were performed. Drynariae Rhizoma extracts treatment reduced the cell viablilty of U937 cells in a dose-dependent manner, which was associated with induction of apoptotic cell death. Drynariae Rhizoma extracts treatment also reduced the levels of Bcl-xL anti-apoptotic protein expression and increased the levels of caspase-3, p53, pro-apoptotic protein, in U937 cells. RT-PCR data revealed that the level of bcl-2, bcl-xL mRNA expressions decreased in a dose-dependent manner. These findings suggest that Drynariae Rhizoma extracts may have induction of apoptotic cell death via regulation of several growth regulatory gene products. The abbreviations used are: FBS, fetal bovine serum; PBS, phosphate buffered saline; PI, propidium iodide; OD, optical density; DiOC6, 3,3-dihexyloxa carbcyanine iodide; MTT, 3 [4-5-dimethylthiazol-2-yl] -2-diphenyltetrazolium bromide.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.1
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• pp.155-165
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• 2006

이 연구는 MCF-7 인간 유방암 세포주에 대한 귀출파징탕(歸朮破癥湯) 추출물의 증식억제효과 세포독성효과 세포사 유발효과를 확인하기 위하여 이루어졌다. MCF-7 인간 유방암 세포주는 Dulbecco's modified Eagle's medium/F12 (DMEM/F12)에 10% fetal bovine serum(FBS;Gibco) 와 항생제를 가하여 만든 배지를 이용하여 배양하였고, MCPF-7 세포를 96- well plate에 접종한 후 다양한 농도(0~2000g/ml)의 귀출파징탕(歸朮破癥湯)이 든 배지로 처리하고 다양한 시간(48, 96, 192)동안 배양하여 현미경으로 관찰하고 각각 MTS assay kit를 이용하여 세포생존율을 측정하였다. 세포독성은 Sulforhodamine B assay 방법을 이용해 측정하였고 세포사 과정에서 MCF-7세포에서의 caspase 활성화를 측정하기 위해 Western blotting을 수행하여 poly ADP ribose polymerase(PARP)의 절단을 확인하였다. 실험결과 귀출파징탕(歸朮破癥湯) 추출물에 의한 세포성장 및 독성효과는 시간 및 농도에 비례하는 것으로 나타났고 세포고사과정에서 작용하는 caspase의 전 기질인 PARP 절단량이 귀출파징탕(歸朮破癥湯) 처리 농도와 시간에 비례하여 증가하였다. 이것은 caspase-3가 MCF-7 세포의 성장을 억제하는데 중요한 역할을 수행함을 의미한다. 따라서 귀출파징탕(歸朮破癥湯)은 다양한 기전에 의해서 유방암 세포에 대한 억제효과를 가진다는 것을 인식할 수 있다.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.1
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• pp.166-177
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• 2006

Purpose : This investigation was undertaken to evaluate the antiproliferation, atoptosis of Sam-nueng(Sparganii Rhizoma) extract using MCF-7 human breast cancer cells. Methods : MCF-7 cells were cultured in Dulbecco's modified Eagle's medium/F12(DMEM/F12) supplemented with 10 % fetal bovine serum(FBS; Gibco) and antibiotics. At varying times after extract treatment, cells were harvested with scraper and processed for analysis of protein expression, proliferation, cytotoxicity and apoptosis. Results : Our results show that the extract of Sam-nueng(Sparganii Rhizoma) strongly inhibits the proliferation of MCF-7 cells in a dose and time-dependent manner. Sulforhodamine B showed that the addition of Sam-nueng(Sparganii Rhizoma) extract reduced the viability of MCF-7 cells in a dose-dependent manner. Poly[ADP(ribose)] polymerase(PARP) which serves as a marker of cells undergoing apoptosis, a major substrate for caspase-3 was extensively cleaved in the Sam-nueng(Sparganii Rhizoma)-treated cells. Conclusion : So, we can conclude that Sam-nueng(Sparganii Rhizoma) can have an inhibitive effect on MCF-7 human breast cancer cells by variable mechanisms.

• The Korean Journal of Physiology and Pharmacology
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• v.11 no.1
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• pp.31-36
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• 2007

To elucidate a potential molecular link between diabetes and atherosclerosis, we investigated the role of Janus tyrosine kinase(JAK) for NAD(P)H oxidase-derived superoxide generation in the enhanced proliferative capacity of vascular smooth muscle cells(VSMC) of Otsuka Long-Evans Tokushima Fatty(OLETF) rat, an animal model of type 2 diabetes. An enhanced proliferative response to 10% fetal bovine serum(FBS) and superoxide generation with an increased NAD(P)H oxidase activity were observed in diabetic(OLETF) VSMC. Both the enhanced proliferation and superoxide generation in diabetic VSMC were significantly attenuated by AG490, JAK2 inhibitor, and PP2, Src kinase inhibitor. Tyrosine phosphorylation of proteins in diabetic VSMC, especially JAK2, was increased compared to control VSMC. Furthermore, the enhanced NAD(P)H oxidase activity in diabetic VSMC was significantly attenuated by AG490 in a dose-dependent manner. Together, these results indicate that the signal pathway which leads to diabetes-associated activation of Src kinase/JAK is critically involved in the diabetic VSMC proliferation through NAD(P)H oxidase activation and superoxide generation.

• Journal of the Korean institute of surface engineering
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• v.50 no.5
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• pp.360-365
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• 2017

In this study, ion release and biocompatibility of sintered Ni-Cr-Ti alloy for dental prosthodontics have been researched by corrosion and cell culture test. The microstructures of the alloys were observed by optical microscope, and corrosion behavior was investigated using potentiostat (Model PARSTAT 2273, EG&G, USA). Cell culture was carried out using hGf cell in DMEM (Welgene Inc., South Korea) supplemented with 10% fetal bovine serum (FBS) (Welgene Inc., South Korea) and antibiotic antimycotic solution (Welgene Inc., South Korea). After corrosion and cell culture test, surface morphologies were observed by field-emission scanning electron microscopy. For wettability behaviors, contact angles were measured by wettability test. As the content of Ti increased, the number of pit decreased and the corrosion resistance was improved from anodic polarization test, also, polarization resistance of samples containing Ti remarkably improved as compared with the alloy not containing Ti. The sintered alloy showed a low contact angle due to the pores formed on the surface. The addition of Ti element showed that the cell survival rate was better than that of the control group.

• International Journal of Oral Biology
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• v.37 no.2
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• pp.43-50
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• 2012

The use of high throughput screening (HTS) in drug development is principally for the selection new drug candidates or screening of chemical toxicants. This system minimizes the experimental environment and allows for the screening of candidates at the same time. Umbilical cord-derived stem cells have some of the characteristics of fetal stem cell and have several advantages such as the ease with which they can be obtained and lack of ethical issues. To establish a HTS system, optimized conditions that mimic typical cell culture conditions in a minimal space such as 96 well plates are needed for stem cell growth. We have thus established a novel HTS system using human umbilical cord derived-mesenchymal stem cells (hUC-MSCs). To determine the optimal cell number, hUC-MSCs were serially diluted and seeded at 750, 500, 200 and 100 cells per well on 96 well plates. The maintenance efficiencies of these dilutions were compared for 3, 7, 9, and 14 days. The fetal bovine serum (FBS) concentration (20, 10, 5 and 1%) and the cell numbers (750, 500 and 200 cells/well) were compared for 3, 5 and 7 days. In addition, we evaluated the optimal conditions for cell cycle block. These four independent optimization experiments were conducted using an MTT assay. In the results, the optimal conditions for a HTS system using hUC-MSCs were determined to be 300 cell/well cultured for 8 days with 1 or 5% FBS. In addition, we demonstrated that the optimal conditions for a cell cycle block in this culture system are 48 hours in the absence of FBS. In addition, we selected four types of novel small molecule candidates using our HTS system which demonstrates the feasibility if using hUC-MSCs for this type of screen. Moreover, the four candidate compounds can be tested for stem cell research application.

• BMB Reports
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• v.42 no.4
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• pp.194-199
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• 2009

The effects of the ginsenoside Rb2 (Rb2) on lipid metabolism were characterized in 3T3-L1 adipocytes to evaluate their utility for treating obesity. While the amounts of total cholesterol and triacylglycerol (TAG) were markedly increased in the adipocytes treated with high amounts of cholesterol and fetal bovine serum (FBS), the test groups treated with Rb2 showed levels that were close to normal. The effect of Rb2 on these cells was comparable to that of lovastatin. Rb2 enhanced the expression of the sterol regulated element binding protein (SREBP) mRNA whereas treatment with cholesterol and FBS led to a reduction in the abundance of this transcript. The activity of fatty acid synthetase (FAS) was lower in the cholesterol group compared to the Rb2 treatment group suggesting that the observed decrease in cholesterol levels and activated SREBP was mediated by Rb2. Treatment with Rb2 also resulted in a decrease in TAG levels in adipocytes cultured under high fatty acid conditions. This effect was mediated by stimulating the expression of SREBP and Leptin mRNA, suggesting that Rb2 might be a valuable component capable of lowering the levels of lipids.