• Journal of Life Science
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• v.25 no.1
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• pp.1-7
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• 2015

Cyclin D is a member of the cyclin protein family, which plays a critical role as a core member of the mammalian cell cycle machinery. D-type cyclins (D1, D2, and D3) bind to and activate the cyclin-dependent kinases 4 and 6, which can then phosphorylate the retinoblastoma tumor suppressor gene products. This phosphorylation in turn leads to release or derepression of E2F transcription factors that promote progression from the G1 to S phase of the cell cycle. Among the D-type cyclins, cyclin D3 encoded by the CCND3 gene is one of the least well studied. In the present study, we have investigated the biochemistry of the proteolytic mechanism that leads to loss of cyclin D3 protein. Treatment of human prostate carcinoma PC-3-M cells with lovastatin and actinomycin D resulted in a loss of cyclin D3 protein that was completely reversible by the peptide aldehyde calpain inhibitor, LLnL. Additionally, using inhibitors for various proteolytic systems, we show that degradation of cyclin D3 protein involves the $Ca^{2+}$-activated neutral protease calpain. Moreover, the half-life of cyclin D3 protein half-life increased by at least 10-fold in PC-3M cells in response to the calpain inhibitor. We have also demonstrated that the transient expression of the calpain inhibitor calpastatin increased cyclin D3 protein in serum-starved NIH 3T3 cells. These data suggested that the function of cyclin D3 is regulated by $Ca^{2+}$-dependent protease calpain.

• Archives of Pharmacal Research
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• v.21 no.3
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• pp.353-356
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• 1998

Geraniol (1), olivetol (2), cannabinoids (3 and 4) and 5-fluorouracil (5) were tested for their growth inhibitory effects against human oral epitheloid carcinoma cell lines (KB) and NIH 3T3 fibrobalsts using two different 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. Cannabigerol (3) exhibited the highest growth-inhibitory activity against the cancer cell lines.

• Journal of Life Science
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• v.24 no.9
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• pp.1001-1005
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• 2014

Ginsenoside Rg3 is one of the active ingredients extracted from red ginseng, and it is an effective chemical component of the human body and well known in herbal medicine as a restorative agent. Several studies have shown that Rg3 has a potent anti-tumor effect on various cancer cell lines. However, Rg3-induced apoptosis in B16F10 melanoma cancer cells is not well understood. In the present study, we tested whether ginsenoside Rg3 could induce apoptosis in B16F10 melanoma cells. We found that Rg3 could inhibit B16F10 melanoma cell viability in a dose-dependent manner, but not normal cells, such as EA.hy.926 and NIH3T3 cells. We also found that Rg3 could induce apoptosis in B16F10 melanoma cells using tunnel-staining assay in a dose-dependent manner. Rg3 treatment induces the phosphorylation of p38 and the expression of Bax, but it inhibits the expressions of the phosphorylation of focal adhesion kinase Bcl2 and pro-caspase3. Taken together, our data suggest that Rg3 could be useful as an anti-cancer agent in B16F10 melanoma cells.

• BMB Reports
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• v.29 no.3
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• pp.205-209
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• 1996

To investigate the role of heparin in keratinocyte growth factor (KGF) and acidic fibroblast growth factor (aFGF) high affinity binding to the KGF receptor (KGFR), a cell free system was established which utilized a secreted chimeric molecule between the KGFR extracellular domain and the immunoglobulin heavy chain Fc domain (KGFR-HFc). KGFR-HFc was purified from NIH 3T3 cells and demonstrated the binding of $[^3H]-heparin$ as well as heparin Sepharose. Scatchard analysis showed that the dissociation constant for heparin binding to KGFR-HFc was 140 nM. High affinity KGF and aFGF binding to KGFR-HFc remained unchanged after treatment with 0.6 M NaCl, which is the concentration sufficient to release any bound heparin to the KGFR-HFc. These results strongly suggest that although the KGFR interacts with heparin, the presence of heparin is not absolutely required for high affinity binding of either KGF or aFGF to the KGFR.

• Journal of Ginseng Research
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• v.19 no.2
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• pp.114-116
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• 1995

Panax ginseng ginsenosides were examined for their affects on the DNA synthesis. The DNA 1 synthesis was measured by the thymidine incorporation into NIH3T3 cells. The ginsenoside, panaxytriol, $Rh_1$ and $Rh_2$ showed reduced [$^{3}H$]-thymidine incorporation. However, other ginsenosides of $Rh_1$, $Rh_2$ and $Rh_3$ did not inhibit DNA synthesis. Among the various ginsenosides, ginsenoside $Rh_2$ was found to be the most inhibitory on DNA synthesis. We suggest $Rh_2$ as one of the potential choice of antiproliferative drugs.

• Journal of Microbiology and Biotechnology
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• v.15 no.3
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• pp.544-550
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• 2005

To develop a highly efficient mammalian expression vector, a series of vectors were constructed based on the murine cytomegalovirus (MCMV) immediate-early (IE) promoter and human ${\beta}$-globin second intron. The resulting MCMV promoter was several-fold stronger than the HCMV promoter in various mammalian cell lines, such as the NIH3T3, Neuro-2a, 293T, and HT1080 cell lines, and was only slightly weaker than the HCMV promoter in HeLa and CHO cells. The inclusion of the human ${\beta}$-globin second intron behind the MCMV promoter or HCMV promoter markedly enhanced the promoter activity in various mammalian cell lines, and the resultant MCMV/Glo-I expression system was stronger than the HCMV promoter from 4.7- to 11.2-fold in every cell line tested. Also, the MCMV/Glo-I promoter induced a higher level of the VSV-G protein in a transiently transfected 293T cell line, which is useful for the production of recombinant retrovirus and lentivirus vectors.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.10
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• pp.1357-1363
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• 2012

Pigs may need to be exploited as xenotransplantation donors due to the shortage of human organs, tissues and cells. Porcine endogenous retroviruses (PERVs) are a significant obstacle to xenotransplantation because they can infect human cells in vitro and have the potential for transmission of unexpected pathogens to humans. In this research, 101 pigs, including four commercial breeds (23 Berkshire, 13 Duroc, 22 Landrace and 14 Yorkshire pigs), one native breed (19 Korean native pigs) and one miniature breed (10 NIH miniature pigs) were used to investigate insertional variations for 11 PERV loci (three PERV-A, six PERV-B and two PERV-C). Over 60% of the pigs harbored one PERV-A (907F8) integration and five PERV-B (B3-3G, B3-7G, 742H1, 1155D9 and 465D1) integrations. However, two PERV-A loci (A1-6C and 1347C1) and one PERV-B locus (B3-7F) were absent in Duroc pigs. Moreover, two PERV-C loci (C2-6C and C4-2G) only existed in Korean native pigs and NIH miniature pigs. The results suggest that PERV insertional variations differ among pig breeds as well as among individuals within a breed. Also, the results presented here can be used for the selection of animals that do not have specific PERV integration for xenotransplantation research.

• Journal of Periodontal and Implant Science
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• v.27 no.3
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• pp.515-524
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• 1997

헤민과 메나디온 제한에 의한 Porphyromonas gingivalis(P. gingivalis) W50의 병독력의 변화를 검색하고자, 실험관내 병독력을 NIH 3T3 세포의 세포활성 변화로 관찰하였고, 생체내 병독력은 배양상청액을 ICR mouse 피하조직에 주사한 후의 염증반응을 관찰하였다. 헤민 존재 하에 배양한 P. gingivalis W50 배양상청액에 의한 mouse 3T3 세포의 세포활성은 헤민과 메나디온 없이 배양한 세포의 활성보다 낮았다. 헤민과 메나디온을 첨가하지 않고 배양한 세균의 생체내 병독력은 중등도의 염증세포 침윤과 울혈에 의한 출혈, 미약한 세포간질의 부종과 근육 파괴를 보였다. 메나디온 존재 하에서 배양한 세균은 미약한 염증세포의 침윤, 울혈에 의한 출혈 및 근육의 파괴가 관찰되었다. 헤민 존재하에서 중등도의 울혈에 의한 출혈, 미약한 세포간질의 부종, 염증세포의 침윤 및 근육파괴가 관찰되었다. 헤민과 메나디온 존재 하에서 배양한 세균은 심한 염증세포의 침윤과 중등도의 세포간질의 부종 및 울혈에 의한 출혈을 보였다. 이상의 연구 결과 P. gingivalis W50 배양 상층액의 병독력은 헤민에 의하여 영향을 받는 것으로 생각된다.

• The Korean Journal of Mycology
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• v.34 no.2
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• pp.105-111
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• 2006

Tremella fuciformis, one of edible and medicinal mushroom belonging to Tremellaceae of Basidiomycota, has been known to have a curative effect on sarcoma 180 of mice and lowering high blood pressure of human beings. Neutral salt soluble [0.9% NaCl (Fr. NaCl)], hot water soluble (Fr. HW) and methanol soluble (Fr. MeOH) substances were extracted from Tremella fuciformis. In vitro cytotoxicity tests, Fr. HW and Fr. NaCl were not cytotoxic against cancer cell lines such as NIH3T3, Sarcoma 180, and HT-29 at the concentration of $2000{\mu}g/ml$, while Fr. MeOH was cytotoxic to NIH3T3 and Sarcoma 180. Intraperitoneal injection with Fr. NaCl showed antitumor effect with life prolongation of 53% in mice inoculated with Sarcoma 180. Fr. NaCl improved the immunopotentiation activity of B lymphocyte by increasing the alkaline phosphatase activity by $3.0{\sim}8.1$ folds, respectively. Intraperitoneal injection with Fr. NaCl increased the numbers of peritoneal exudated cells and circulating leukocytes by 7.4 folds and 1.6 folds, respectively, than in the control group. The antitumor effect of T. fuciformis against Sarcoma 180 of mice was likely due to immunopotentiation activity.

• Molecules and Cells
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• v.27 no.6
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• pp.697-701
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• 2009

We previously identified cancer-upregulated gene 2 (CUG2) as a commonly up-regulated gene in various human cancer tissues, especially in ovary, liver, and lung (Lee et al., 2007a). CUG2 was determined to be a nuclear protein that exhibited high proto-oncogenic activities when overexpressed in NIH3T3 mouse fibroblast cells. To identify other cellular functions of CUG2, we performed yeast two-hybrid screening and identified CENP-T, a component of CENP-A nucleosome complex in the centromere, as an interacting partner of CUG2. Moreover, CENP-A, the principle centromeric determinant, was also found in complex with CENP-T/CUG2. Immunofluorescent staining revealed the co-localization of CUG2 with human centromeric markers. Inhibition of CUG2 expression drastically affected cell viability by inducing aberrant cell division. We propose that CUG2 is a new component of the human centromeric complex that is required for proper chromosome segregation during mitosis.