• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.1 s.32
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• pp.169-176
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• 2007

Objectives : The purpose of this study was to investigate inhibitory effect of Gagamtongsung-San(GTS) on the cancer. Methods : This study estimated the cytotoxicity of GTS about L1210 and NIH3T3. We used GTS extract distilled with water, n-Hexane, Ethyl acetate and Butanol. The cytotoxicitys of GTS about cancer cells and normal cells were tested using a colorimetric tetrazoliun assay(MTT assay). Results : The results of this study were obtained as follow ; l. Cytotoxicity of water extract of GTS in L1210 cell lines was significantly increased, compared with NIH3T3. 2. n-Hexane fraction of GTS had similar cytotoxicity between L1210 and NIH3T3, and that have similar $IC_{50}$ of water extract of GTS at 276 ${\mu}g/ml$ 3. Ethyl acetate fraction of GTS had low degree cytotoxicity both L1210 and NIH3T3 cell lines. 4. Butanol fraction of GTS had cytotoxicity between L1210 and NIH3T3. Significantly, Cytotoxicity of GTS in L1210 cell lines was significant increased. 5. $H_2O$ fraction of GTS had no cytotoxicity both L1210 and NIH3T3.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.1 s.32
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• pp.145-153
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• 2007

Objecgtive : The aim of present study was to investigate inhibition effect of Whakijogyung-Tang(WJT) on the tumor cell lines. This study estimated the cytotoxicity of WJT about viability of S-180 and NlH3T3. Methods : The cytotoxicity of WJT about viability of cells were tested using a colorimetric tetrazoliun assay(MTT assay) Results and Conclusion : 1. Water extract of WJT had $IC_{50}$ of 863 ${\mu}g/ml$ in S-180 cell lines, but cytotoxicity of NIH3T3 was not significant difference compare with S-180. 2. n-Hexane fraction of WJT had similar cytotoxicity between S-180 and NIH3T3, but that could not have $IC_{50}$ in S-180 cell lines. 3. Ethyl acetate fraction of WJT had low degree cytotoxicity both S-180 and NIH3T3 cell lines. 4. Significantly, Butanol fraction of WJT had differenct citotoxicity between S-180 and NIH3T3. 5. $H_2O_2$ fraction of WJT had no cytotoxicity both S-180 and NIH3T3.

• Korean Journal of Veterinary Research
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• v.30 no.4
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• pp.459-464
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• 1990

Inhibitory effect of caffeine on NIH3T3 cell proliferation was studied by using [$^3H$]thymidine incorporation and a modified one-step silver staining technique. The latter technique shows argyrophilic nucleolar organizer region-associated proteins (Ag-NORs), which are seen in nuclei as black dots. The result was compared with the counts of [$^3H$] thymidine incorporation. The results were summarized as follows; 1. The Ag-NORs numbers of NIH3T3 cells were $6.81{\pm}1.38$ at 24 hrs, $7.13{\pm}1.26$ at 48 hrs, $8.50{\pm}2.04$ at 72 hrs after incubation. 2. The numbers of Ag-NORs were significantly decrease in caffeine treated groups (p<0.01). 3. The counts of [$^3H$] thymidine incorporation were similar to the result of using Ag-NORs staining technique. It is concluded that Ag-NOR is a rapid, simple and compatible method to quantitate cell proliferation as compared with [$^3H$]thymidine incorporation.

• Molecules and Cells
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• v.27 no.3
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• pp.299-305
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• 2009

Over-expression of phospholipase D (PLD) 1 or PLD2 down-regulated CKII activity in NIH3T3 cells. The same results were found with catalytically inactive mutants of PLD isozymes, indicating that the catalytic activity of PLD is not required for PLD-mediated CKII inhibition. Consistent with this, 1-butanol did not alter CKII activity. The reduction in CKII activity in PLD-over-expressing NIH3T3 cells was due to reduced protein level, but not mRNA level, of the $CKII{\beta}$ subunit. This PLD-induced $CKII{\beta}$ degradation was mediated by ubiquitin-proteasome machinery, but MAP kinase and mTOR were not involved in $CKII{\beta}$ degradation. PLD isozymes interacted with the $CKII{\beta}$ subunit. Immunocytochemical staining revealed that PLD and $CKII{\beta}$ colocalize in the cytoplasm of NIH3T3 cells, especially in the perinuclear region. PLD binding to $CKII{\beta}$ inhibited $CKII{\beta}$ autophosphorylation, which is known to be important for $CKII{\beta}$ stability. In summary, the current data indicate that PLD isozymes can down-regulate CKII activity through the acceleration of $CKII{\beta}$ degradation by ubiquitin-proteasome machinery.

• The Journal of Natural Sciences
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• v.15 no.1
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• pp.89-95
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• 2005

JSRV, which causes sheep lung cancer, is known to have the transforming activity of NIH3T3 cells. Especially Envelope protein of this virus has the transforming activity to NIH3T3. To know the effects on the transcriptional activity of transcription factors by this viral protein transient transfection was performed by using the luciferase reporter system. The result showed that JSRV Envelope protein increased the transcriptional activity of NF-kB and AP-1.

• Proceedings of the Korean Society of Toxicology Conference
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• 2002.05a
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• pp.81-81
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• 2002

The recent development of cDNA microarray or cDNA chip technology has made it possible to analyze the expression of thousands of genes at once. In present study, we made radioresistant clones (#1 and #4) from NIH3T3 cells which are not tumorigenic and we identified 4 genes using microarray system, cdk6, cdc25B, mdm-2 and nidogene, which were altered in radiaiton resistanct NIH3T3 cells.(omitted)

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.4
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• pp.633-640
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• 2004

This study was performed to investigate the cytotoxic effect of the water-extract from Chaga mushroom (Inonotus obliquus) compositions containing powdered green tea in HCT-15 human colon carcinoma, AGS human gastric carcinoma and NIH3T3 mouse normal fibroblast cells using viable cell count and MTT assay. The water-extract from Chaga mushroom compositions induced inhibitory effects on proliferation of HCT-15 and AGS cells in the MTT assay and viable cell count. However, mouse normal NIH3T3 cells were exhibited 80% survival under the same condition. Chaga mushroom compositions showed highly antiproliferative effect in human cancer cell line HCT-15 and AGS, but not in mouse normal cell line NIH3T3. These results suggest that Chaga mushroom (Inonotus obliquus) compositions containing powdered green tea are the candidate for chemoprevention in colon and gastric cancer.

• The Korea Journal of Herbology
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• v.21 no.1
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• pp.101-107
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• 2006

Objectives : It is well known that hexavalent chromium has toxic effect on normal cells. Recently, toxic effect of hexavalent chromium is diminished by the some extracts derived from herbs or plants. But, the toxic or protective mechanism of hexavalent chromium is well unknown. This study was performed to examine the protective effect of poncirin against $Na_2Cr_2O_7$-induced cytotoxicity on NIH3T3 fibroblasts. Methods : The protective effect of the cytotoxicity induced by $Na_2Cr_2O_7$ was measured by the cell viability after NIH3T3 fibroblasts were cultured with or without $Na_2Cr_2O_7$ for 48 hours. Antitoxic effects of poncirin on the cytotoxicity induced by $Na_2Cr_2O_7$ were examined by colorimetric assays such as MTT or XTT assay. Results : $Na_2Cr_2O_7$ decreased cell viability by the decreased absorbance in MTT or XTT assay, but, the poncirin increased cell viability which was decreased by $Na_2Cr_2O_7$-induced cytotoxicity on NIH3T3 fibroblasts. Conclusion : These results suggest that $Na_2Cr_2O_7$ showed cytotoxicity effect on NIH3T3 fibroblasts by the decrease of cell viavility, and poncirin was effective in the protection of $Na_2Cr_2O_7$-induced cytotoxicity in these cultures.

• Applied Microscopy
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• v.36 no.3
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• pp.165-172
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• 2006

Secretory leukocyte protease inhibitor (SLPI) involves tissue protection against the destructive action of neutrophil elastase at the site of inflammation. Several studies on new functions of SLPI have demonstrated that SLPI may play a primary role in innate immunity than protease inhibitor, To identify the function of SLPI by lipopolysaccharide (LPS) stimulation in the embryonic fibroblast (NIH3T3) cells. we studied the expression of SLPI compared to other growth factors involving the LPS treatment. To address this, we performed the reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of the SLPI and some growth factors such as VEGF. bFGF, and PDGF-BB after LPS stimulation. NIH3T3 cells were exposed 100 ng/mL Escherichia coli LPS for 30min, 60min, 90min, 24h, and 48h, respectively. The result of RT-PCR showed that SLPI and VEGF mRNA was expressed strongly in NIH3T3 without related to LPS stimulation. mRNA of bFGF was weakly expressed such as the expression of the control. PDGF mRNA expression gradually increased follows at time course. However, SLPI protein level was increased in lysates and culture medium by LPS stimulation. Phase contrast microscopic and scanning electron microscopic observation showed that the increased cell number and cytoplasmic enlargement of the NIH3T3 cells. Therefore, it suggests that the LPS upregulates SLPI expression in NIH3T3 cells. Moreover, secreted SLPI may stimulate cell proliferation and migration.

• Microbiology and Biotechnology Letters
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• v.39 no.4
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• pp.337-344
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• 2011

Integrase MJ1 from the bacteriophage ${\Phi}FC1$ carries out recombination between two DNA sequences (the phage attachment site, attP and the bacterial attachment site, attB) in NIH3T3 mouse cells. In this study, the integration vector containing attP, attB and the integrase gene MJ, was constructed. The integration mediated by integrase MJ1 in Escherichia coli led to excision of LacZ. Therefore, the frequency of integration was measured by the counting of the white colony, which is detectable on X-Gal plates. The extrachromosomal integration in NIH3T3 mouse cells was monitored by the expression of the green fluorescent protein (GFP) as a reporter. To demonstrate integration mediated integrase MJ1 in NIH3T3 cells, vectors containing attP and attB were co-transfected into NIH3T3 cells. The integration was confirmed by fluorescent microscopy. The expression of GFP was induced in NIH3T3 cells expressing MJ1 without accessory factors. By contrast, the excision mediated by the MJ1 between attR and attL had no effect on the expression of GFP. These results suggest that integrase MJ1 may enable a variety of genomic modifications for research and therapeutic purposes in higher living cells.