• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6863-6867
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• 2013

Phyllanthus emblica (PE) is known to exhibit various pharmacological properties. This study aimed to evaluate the antimetastatic potential of a PE aqueous extract. Cytotoxicity to human fibrosarcoma cells, HT1080, was determined by viability assay using the 3-(4,5-dimethylthiazol,2-yl)-2,5-diphenyltetrazolium bromide (MTT) reagent. Cell migration and invasion were investigated using chemotaxis chambers containing membranes precoated with collagen IV and Matrigel, respectively. Cell attachment onto normal surfaces of cell culture plates was tested to determine the cell-adhesion capability. The molecular mechanism of antimetastatic activity was assessed by measuring the gene expression of matrix metalloproteinases, MMP2, and MMP9, using reverse transcription-polymerase chain reaction (RT-PCR) assay. The mRNA levels of both genes were significantly down-regulated after pretreatment with PE extract for 5 days. Our findings show the antimetastatic function of PE extract in reducing cell proliferation, migration, invasion, and adhesion in both dose- and time-dependent manners, especially growth arrest with low $IC_{50}$ value. A decrease in the expression of both MMP2 and MMP9 seems to be the cellular mechanism for antimetastasis in this case. There is a high potential to use PE extracts clinically as an optional adjuvant therapeutic drug for therapeutic intervention strategies in cancer therapy or chemoprevention.

• Journal of Microbiology and Biotechnology
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• 제15권3호
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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.

• 혜화의학회지
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• 제8권1호
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• pp.131-146
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• 1999

To evaluate the antitumor activity of Kamisagoonjatang(KST), Kami-jihwangtang (KJT) and Kamigoonjajihwangtang(KKJT), studies were done experimentally. The results were obtained as follows: 1. In cytotoxicity against B16-F10, HT1080, SNU, and L1210, con-centration inhibiting cell growth up to below 50% of control was recognized at $10^{-3}g/ml$ of KKJT. 2. In cytotoxicity against A549, SK-OV-3, XF498 and HCT15, concentration inhibiting cell growth up to below 30% of control was over $400{\mu}g/ml$ of KKJT only and also over $200{\mu}g/ml$ against SK-MEL-2. 3. In Inhibitory effect on activity of DNA topoisomerase I, the $IC_{50}$ was shown $200-400{\mu}g/m{\ell}$, of KST, over $400{\mu}g/m{\ell}$ of KJT and $100-200{\mu}g/m{\ell}$ of KKJT. 4. The T/C% was 122.8 in KJT, 127.4 in KST and 158.4 in KKJ-Ttreated group in S-180 bearing ICR mice. 5. In hematological changes in S-180 bearing ICR mice, numbers of WBC were decreased significantly in KJT and KKJT treated groups as compared with control, whereas those of platelet were increased with no significance in all groups as compared with control. From above results it was concluded that KKJT could be usefully applied for the prevention and treatment of cancer.

• 혜화의학회지
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• 제9권1호
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• pp.155-167
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• 2000

To evaluate the antitumor activity and antimetastatic effects of Gobongeerlhobang(GG1B), studies were done experimentally. The results were obtained as follows: 1. GG1B extracts exhibited a significant cytotoxicity against HT1080, A549, SK-MEL-2 and SK-OV-3 cell lines. 2. The T/C% was 120.7% in GG1B treated group in S-180 bearing ICR mice. 3. GG1B extracts exhibited inefficient adhesive effect of A549, B16-BL6 cell to complex extracellular matrix. 4. GG1B extracts showed a significant inhibition of lung metastasis of B16-BL6 cells in C57BL/6. 5. In CAM assays, angiogenesis was insignificantly inhibited in GG1B treated group than control group. These results suggested that GG1B extracts might be usefully applied for prevention and treatment of cancer.

• BMB Reports
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• 제47권7호
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• pp.393-398
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• 2014

The role of Bax inhibitor-1 (BI-1) in the protective mechanism against apoptotic stimuli has been studied; however, as little is known about its role in death receptor-mediated cell death, this study was designed to investigate the effect of BI-1 on Fas-induced cell death, and the underlying mechanisms. HT1080 adenocarcinoma cells were cultured in high concentration of glucose media and transfected with vector alone (Neo cells) or BI-1-vector (BI-1 cells), and treated with Fas. In cell viability, apoptosis, and caspase-3 analyses, the BI-1 cells showed enhanced sensitivity to Fas. Fas significantly decreased cytosolic pH in BI-1 cells, compared with Neo cells, and this decrease correlated with BI-1 oligomerization, mitochondrial $Ca^{2+}$ accumulation, and significant inhibition of sodium-hydrogen exchanger (NHE) activity. Compared with Neo cells, a single treatment of BI-1 cells with the NHE inhibitor EIPA or siRNA against NHE significantly increased cell death, which suggests that the viability of BI-1 cells is affected by the maintenance of intracellular pH homeostasis through NHE.

• 동아시아식생활학회지
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• 제11권1호
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• pp.19-25
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• 2001

This study was performed to examine the antimutagenic and cytotoxic effects of potato vinegar and commercial vinegars(cider, brown rice, persimmon vinegars) on Salmonella typhimurium TA98, TA100 and cancer cell lines using Ames test and cytotoxicity assay, respectively. In Ames test, all vinegars did notexhibit any mutagenicity , but showed substantial inhibitory effects against N- methyl - N -nitro - N- nitrosog -uanidine(MNNG) , 4-nitroquinoline-1-oxide(4NQO), 3-amino-1,4-dimethyl-5H-pyrido(4,3-b)indol(Trp-P-1)and benzo( $\alpha$ )pyrene(B( $\alpha$ )P). The number of revertants per plate decreased significantly when these vinegars(80 ug/plate) were added to the assay system using TA100 strain. Especially, potato vinegar(80 ug/plate) showed high inhibition rate of 69.9% against mutagenicity of B( $\alpha$ )P on TA100 strain. In the cytotoxicity assay, these vinegars also showed prominent cytotoxic activity against human cancer cell lines. Potato vinegar(10 ug/well) showed the strongest cytotoxic effect against HT1080 (fibrosacoma cell) andK562 ( myelogenous leukemia) at the same concentration when compared with other vinegars.

• Natural Product Sciences
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• 제13권2호
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• pp.128-131
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• 2007

Tumor angiogenesis is a critical step f3r the growth and metastasis of solid tumors. Vascular endothelial growth factor (VEGF) is the most important angiogenic molecule associated with tumor-induced neovascularization. VEGF exerts its activity through binding to its receptor tyrosine kinase, KDR/Flk-1, expressed on the surface of endothelial cells. This study was carried out to investigate inhibitory effect of extracts from root cortex of Paeonia suffruticosa Andrews on VEGF binding to VEGF receptor. The MeOH extract from P. suffrutiocosa Andr. inhibited the binding of KDR/Flk-1-Fc to immobilized VEGF$_{165}$ more than 45% at the concentration of 100 ${\mu}$g/mL. The MeOH extract was further fractionated into n-hexane, ethyl acetate, n-BuOH, and aqueous fractions. Among the four fractions, the ethyl acetate fraction from the root cortex of P. suffruticosa Andr. exhibited highly effective inhibition (${\approx}$ 79% inhibition) and then n-BuOH fraction (${\approx}$ 45% inhibition) on the binding of KDR/Flk-1-Fc to immobilized VEGF$_{165}$ at the concentration of 100 ${\mu}$g/mL. The ethyl acetate fraction from the root cortex of P. suffruticosa Andr. more efficiently blocked VEGF-induced human umbilical vein endothelial cell proliferation, than the growth of HT1080 human fibrosarcoma. Our results suggest that P. suffruticosa Andr. may be used as a candidate fur developing anti-angiogenic agent.

• Molecules and Cells
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• 제38권1호
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• pp.33-39
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• 2015

The correction of disease-causing mutations by single-strand oligonucleotide-templated DNA repair (ssOR) is an attractive approach to gene therapy, but major improvements in ssOR efficiency and consistency are needed. The mechanism of ssOR is poorly understood but may involve annealing of oligonucleotides to transiently exposed single-stranded regions in the target duplex. In bacteria and yeast it has been shown that ssOR is promoted by expression of $Red{\beta}$, a single-strand DNA annealing protein from bacteriophage lambda. Here we show that $Red{\beta}$ expression is well tolerated in a human cell line where it consistently promotes ssOR. By use of short interfering RNA, we also show that ssOR is stimulated by the transient depletion of the endogenous DNA mismatch repair protein MSH2. Furthermore, we find that the effects of $Red{\beta}$ expression and MSH2 depletion on ssOR can be combined with a degree of cooperativity. These results suggest that oligonucleotide annealing and mismatch recognition are distinct but interdependent events in ssOR that can be usefully modulated in gene correction strategies.

• 치과방사선
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• 제29권2호
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• pp.435-449
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• 1999

Purpose : The study was aimed to detect the differences in the cell viability and the apoptosis induction after irradiation on normal and tumorigenic cells. Materials and Methods : The study. that was generated for two human normal cells(RHEK, HGF-l) and two human tumor cells(KB. HT-1080). was tested using MTT assay at 1 day and 3 day after irradiation and TUNEL assay under confocal laser scanning microscope at 1 day after irradiation. Single irradiation of 0.5. 1, 2. 4. and 8Gy were applied to the cells. The two fractions of 1. 2. 4. and 8Gy were separated with a 4-hour time interval. The irradiation was done with 5.38Gy/min dose rate using Cs-137 irradiator at room temperature. Results and Conclusions : 1. In 3-day group. the cell viability of HGF-1 cell was significantly decreased at 2. 4 and 8Gy irradiation, the cell viability of KB cell was significantly decreased at 8Gy irradiation and the cell viability of HT-I080 cell was significantly decreased at 4 and 8Gy irradiation. 2. There was significant difference between RHEK and KB cell line in the cell viability of 3-day group at 8Gy irradiation. There was significant difference between RHEK and HGF-1 cell line in the cell viability of 3-day group at 4 and 8Gy irradiation. 3. There was a significantly decreased cell viability in 3-day group than those in 1-day group at 2. 4 and 8Gy on HGF-1 cell. at 4 and 8Gy on HT-I080 cell. at 8Gy on KB cell. 4. We could detect DNA fragmented cells only on KB cell. Number of apoptotic cells of KB cell was significantly increased at 4 and 8Gy irradiation. However, there was no correlation between cell viability and apoptosis. 5. On all 4 cell lines, there were no differences between single and split irradiation method in cell viability and apoptosis.

• 생명과학회지
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• 제19권11호
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• pp.1598-1604
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• 2009

양파 껍질의 폐기는 생리활성 물질이 손실 될 수 있으므로 양파 껍질을 이용한 연구는 폐자원을 활용한다는 면에서 의미가 있다. 이에 본 연구에서는 양파를 육질과 껍질로 나누어 이들의 총 플라보노이드 함량을 측정하였고 세포 내 활성산소종 제거 및 인체 암세포 증식 억제 효과에 대하여 비교 검토하였다. 양파 껍질은 육질과 비교했을 때 48배의 플라보이드 함량을 나타내어 양파 껍질에 플라보노이드 성분이 다량 함유되어 있음을 확인하였다. 양파 껍질의 A+M과 MeOH 추출물을 0.05 mg/ml의 농도로 HT-1080 세포에 처리하였을 때 지질 과산화물의 생성을 크게 억제시켰다. 이들 두 추출물들은 $500{\mu}M$ $H_2O_2$만을 처리한 control군과 비교하였을 때 측정 시간 120분 동안 계속적으로 높은 세포 내 지질 과산화물 생성 억제능력을 나타내었고 특히 양파 껍질에 의한 저해 효과가 양파 육질보다 높았다. 양파 육질과 껍질의 추출물로부터 얻어진 분획물들 중에서는 양파 껍질의 85% aq. MeOH 및 BuOH 분획물에 의한 저해효과가 우수하였고 양파 육질의 경우에는 85% aq. MeOH 분획물에 의한 항산화효과가 높았다. 암세포증식 억제 실험에서 양파 껍질의 A+M 추출물의 $IC_{50}$은 AGS 및 HT-29 세포에서 각각 0.07 및 0.05 mg/ml로 양파 육질보다 저해효과가 높았고 껍질 MeOH 추출물의 $IC_{50}$은 두 세포에서 각각 0.75 및 3.31 mg/ml로 AGS 세포에 대한 증식 억제효과가 우수하였음을 관찰 할 수가 있었다. 분획물들 중에서는 양파 육질 및 껍질의 85% aq. MeOH 분획물의 AGS 세포에 대한 $IC_{50}$이 각각 0.04 및 0.03 mg/ml로 가장 높은 활성을 나타내었고. 이들 분획물들도 HT-29 세포보다는 AGS 세포에 대한 증식 억제효과가 높았음을 살펴 볼 수가 있었다. 대체로 water 분획물에 의한 활성산소종 저해 및 암세포 증식 억제효과는 분획물들 중 가장 낮았다. 이상의 결과로부터 양파 부산물인 껍질에도 생리활성 물질이 함유되어 있는 것으로 판단되므로 이를 활용한 양파가공품 개발이 필요하다고 여겨진다.