• Biomolecules & Therapeutics
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• 제6권4호
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• pp.345-350
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• 1998

The adverse effects of ethanol on cell proliferation have been described for a variety of tissues and cells. In the present study, we investigated whether chronic ethanol intoxication impairs the cell proliferation and DNA synthesis induced by oncogenic $H-ras^{V12}$ - and $v-K-ras^{V12}$-transformed cells. Ethanol treatment inhibited the cell proliferation and the DNA synthesis of control parental fibroblasts in a time- and dose-dependent manner. In contrast, ethanol did not suppress the proliferation of either oncogenic $H-ras^{V12}$ - or $v-K-ras^{V12}$ -transformed fibroblasts. Microinjection of oncogenic $H-Ras^{V12}$ protein induces DNA synthesis and ethanol treatment did not interfere with the DNA synthesis. The antiproliferative toxicity of ethanol was rescued by antioxidants, such as N-acetylcysteine and 4-methlpyrazole. These results indicate that the antiproliferative action site of ethanol toxicity lies upstream or is independent of Ras and ethanol exerts its toxicity through a free radical formation.

• 생명과학회지
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• 제17권12호
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• pp.1723-1728
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• 2007

본 연구에서는 다양한 기능성을 가지는 복분자를 80% methanol로 추출한 후 분획물들을 제조하여 항산화 효과 및 인간 유래 위암세포인 AGS와 KatoIII을 이용하여 성장 저해 효과와 DNA손상을 측정하였다. 항산화 효과를 측정해 보기 위하여 총 폴리페놀 함량과 플라보노이드 함량, DPPH free radical의 소거활성을 측정하였는데, 먼저 폴리페놀의 경우 butanol fr., ethylacetate fr., water fr.순으로 나타났고 특히 butanol fr.의 경우 $546.25{\mu}g/mg$의 매우 높은 폴리페놀 함량을 확인 할 수 있었다. 플라보노이드의 함량 결과 ethylacetate fr., butanol fr., water fr.,순으로 나타났으며 ethylacetate fr.의 경우 $141.78{\mu}g/mg$의 높은 플라보노이드 함량을 확인할 수 있었다. DPPH free radical의 소거활성에서는 폴리페놀 함량의 순서와 같은 효과를 보였으며, 합성 항산화제인 BHA의 $RC_{50}$값 $3{\mu}g/ml$보다 water fr.을 제외하고 더 좋은 소거능을 확인 할 수 있었다. 인간 유래 위암세포주의 성장 저해 효과를 살펴본 결과 AGS와 KatoIII 세포주 모두 butanol fr.에서 가장 좋은 저해능을 나타내었고, butanol fr., ethylacetate fr., water fr. 순서로 저해효과를 나타내었다. DNA손상을 측정한 결과 역시 이와 같은 순으로 나타났으며, butanol fr.에서 1시간 이내에 약 50%정도의 손상유도 효과를 보였다. 이상의 결과에서 복분자 methanol추출물과 각각의 분획물은 항산화 효과 및 위암 세포주에 대한 성장 저해 활성 및 높은 DNA손상 유도 효과를 갖고 있음을 알 수 있었다.

• 한국가축번식학회지
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• 제19권2호
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• pp.89-93
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• 1995

In this study was demonstrate that retrovirus-mediated gene transfer is one of the promising alternatives to the conventional pronuclear DNA microinjection approach, especially in transferring the exogenous genes into the boving embryos. By co-culturing of zona of zona-free one-cell stage embryos with the retrovirus-producing cells for 24 hours followed by 6 days of culture in virus-free medium, we could get morulae and blastocysts expressing the E. coli LacZ genes which were transferred by our retrovirus vector. The results obtained in this study are summarized as follows : 1. Addition of 5$\mu\textrm{g}$/ml of polybrene in the embryo and virus-producing cell co-culture medium did not affect development of zona-free one-cell embryo. 2. Compared with the intact embryos removal of zona at one-cell stage before co-culturing with the virus-producing cells for one day caused only slight decrease of embryo develpment. 3. Co-culture of 625 zona-free one-cell stage embryos with the virus-producing cells resulted in 65(10.4%) morulae or blastocysts, and 12.3%(8/65) of the morulae or blastocysts were E. coli LacZ positive.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3281-3285
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• 2012

Objectives: To evaluate the clinicopathologic correlation and prognostic value of HPV18 DNA viral load in patients with early-stage cervical neuroendocrine carcinoma (NECA). Methods: Formalin-fixed, paraffin-embedded tissue of cervical NECA patients with known HPV18 infection and clinicopathologic data including follow-up results were collected. The HPV18 DNA load was assessed with quantitative PCR targeting the HPV18 E6E7 region. Results: Twenty-one patients with early-stage (IB-IIA) cervical NECA were identified. HPV18 DNA viral load ranged from 0.83 to 55,174 copies/cell (median 5.90). Disease progression, observed in 10 cases (48%), was not significantly associated with any clinicopathologic variables. However, the group of patients with progressive disease tended to have a higher rate of pelvic lymph node metastasis (50% versus 9%, p=0.063) and a lower median value of HPV18 DNA viral load (4.37 versus 8.17 copies/cell, p=0.198) compared to the non-recurrent group. When stratified by a cut-off viral load value of 5.00 copies/cell, the group of patients with viral load ${\leq}5.00$ copies/cell had a significantly shorter disease-free survival than the group with viral load >5.00 copies/cell (p=0.028). The group with a lower viral load also tended to have a higher rate of disease progression (75% versus 31%, p=0.080). No significant difference in the other clinicopathologic variables between the lower and higher viral load groups was identified. Conclusion: HPV18 DNA viral load may have a prognostic value in patients with early-stage NECA of the cervix. A low viral load may be predictive of shortened disease-free survival in these patients.

• Journal of Acupuncture Research
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• 제15권2호
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• pp.135-145
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• 1998

Bupleury radix has been used for the treatment of fever, liver disease, inflammation in traditional medicine. The present study was carried out to evaluate the antioxidant effects of Bupleury radix aqua-acupuncture solution (BRAS) in vitro. Oxygen derived free radicals produced in the course of normal aerobic life, such as superoxide anion radical($O_2^-$ ), hydroxyl radicaI( OH), hydrogen peroxide($H_2O_2$) and singlet oxygen($^1O_2$) can attack polyunsaturated fatty acid in cell membranes, enzymes, other cell compounds, and give rise to lipid peroxidation, DNA damage, lipofuscin accumulation, structure alteration of cell membrane and cell death. In this study, antioxidant effects of BRAS on lipid peroxidation were determined according to the method of TBA. BRAS inhibited markedly peroxidation of linoleic acid during the autoxidation, and also inhibited lipid peroxidation induced by hydroxyl radical derived from $H_2O_2-Fe^{2+}$ in rat liver homogenate. And BRAS showed 30% scavenging effect on DPPH radical, also exhibited a 30% inhibitory effect on superoxide generation from xanthine-xanthine oxidase system. In addition, BRAS protected the cell death induced by tert-butyl hydroperoxide(t-BHP) and significantly increased cell viability in the normal rat liver cell(Ac2F).

• BMB Reports
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• 제32권3호
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• pp.266-272
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• 1999

The insulin-like growth factor (IGF) system consisting of IGF-I, IGF-II, IGF-receptors, and IGF-binding proteins (IGFBP) regulates the proliferation of a variety of cancer cell types. To examine whether a decrease in endogenous IGFBP-3 stimulates proliferation or inhibits differentiation, Caco-2 cells, a human colon adenocarcinoma cell line, were stably transfected with an anti-sense IGFBP-3 expression construct or pcDNA3 vector as control. Accumulation of IGFBP-3 mRNA and secretion of IGFBP-3 into serum-free conditioned medium, 9 days after plating, were significantly lower in Caco-2 cell clones transfected with anti-sense IGFBP-3 cDNA compared to the controls. The anti-sense clones grew at a similar rate to the controls for 8 days after plating, but achieved a higher final density between days 10 and 12. The levels of sucrase-isomaltase mRNA, a marker of enterocyte differentiation of Caco-2 cells, were lower in the anti-sense clones examined on day 9. In conclusion, proliferation of Caco-2 cells can be stimulated by lowering endogenously-produced IGFBP-3.

• 한국식품위생안전성학회지
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• 제17권2호
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• pp.106-116
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• 2002

양강추출물 (Alpinia officinarum, 70% ethanol extract)과 함유 주성분인 galangirn은 프리라디칼소거작용과 지질과산화억제활성을 나타내었으며, $H_2O$$_2$ 또는 KO$_2$유도 세포독성에 대해서도 억제적으로 작용하여 세포보호효과를 나타내었다. DNA single strand breakage와 같은 산화적 스트레스에 대해서도 보호작용을 하고 있다. 또한, 마우스 소핵시험에 의하여 adriamycin과 같은 superoxide 우발물질에 의한 염색체 수준에서의 손상에 대해서도 억제효과를 나타내었다. 양강추출물은 galangin을 유효성분으로하여 산소프리라디칼들에 의한 산화적 손상에 억제적으로 작용하는 기전으로 산화적 스트레스에 대한 항노화, 암예방제로서의 응용가능성이 높은 추출물로서 향후 기능성 식품으로서의 응용가능성이 높을 것으로 판단되었다.

• Preventive Nutrition and Food Science
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• 제10권1호
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• pp.46-51
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• 2005

The protective effect of Acanthopanax senticosus (AS) extract on alloxan-induced pancreatic β-cell damage was investigated in HIT T-15 cells, a Syrian hamster pancreatic β-cell line. Alloxan caused the pancreatic β-cell damage through the generation of reactive oxygen free radicals, increased DNA fragmentation, and decreased cellular NAD/sup +/ levels. The β-cell damage was significantly prevented by the pretreatment with water soluble extract of AS roots. These results suggest that the protective effect of AS extract, on alloxan-induced β-cell damage, is primarily due to the inhibition of the generation of reactive oxygen free radical species (ROS) by alloxan.

• Asian Pacific Journal of Cancer Prevention
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• 제17권3호
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• pp.939-944
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• 2016

Background: Cell-free DNA circulating in blood is a candidate biomarker for malignant tumors. Unlike uniformly truncated DNA released from apoptotic non diseased cells, DNA released from necrotic cancer cells varies in size. Objectives: To measure the DNA integrity index in serum and the absolute DNA concentration to assess their clinical utility as potential serum biomarkers for colorectal carcinoma (CRC) compared to CEA and CA19-9. Materials and Methods: Fifty patients with CRC, 10 with benign colonic polyps and 20 healthy sex and age matched volunteers, were investigated by real time PCR of ALU repeats (ALU q-PCR) using two sets of primers (115 and 247 bp) amplifying different lengths of DNA fragments. The DNA integrity index was calculated as the ratio of q-PCR results of ALU 247/ALU 115bp. Results: Serum DNA integrity was statistically significantly higher in CRC patients compared to the benign and control groups (p<0.001). ROC curves for differentiating CRC patients from normal controls and benign groups had areas under curves of 0.90 and 0.85 respectively. Conclusions: The DNA integrity index is superior to the absolute DNA concentration as a potential serum biomarker for screening and diagnosis of CRC. It may also serve as an indicator for monitoring the progression of CRC patients. Combining CEA and CA19-9 with either of the genetic markers studied is better than either of them alone.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5563-5568
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• 2012

Based on our previous report on gastric cancer which documented ATP4A and ATP4B mRNA down-regulation in gastric tumors relative to normal gastric tissues, we hypothesized that epigenetic mechanisms could be responsible. ATP4A and ATP4B mRNA expression in gastric cancer cell lines AGS, SNU638 and NUGC-3 was examined using reverse transcriptase PCR (RT-PCR). AGS cells were treated with TSA or 5'-AzaDC and methylation specific PCR (MSP) and bisulfite sequencing PCR (BSP) analysis were performed. MSP analysis was on DNA from paraffin embedded tissues sections and plasma. Expression analysis revealed downregulation of ATP4A and ATP4B genes in gastric cancer cell lines relative to normal gastric tissue, while treatment with 5'-AzaDC re-activated expression of both. Search for CpG islands in their putative promoter regions did not indicate CpG islands (CGI) but only further downstream in the bodies of the genes. Methylation specific PCR (MSP) in the exon1 of the ATP4B gene and exon7 in ATP4A indicated methylation in all the gastric cancer cell lines tested. MSP analysis in tumor tissue samples revealed methylation in the majority of tumor samples, 15/19, for ATP4B and 8/8 for ATP4A. There was concordance between ATP4B and ATP4A down-regulation and methylation status in the tumour samples tested. ATP4B methylation was detectable in cell free DNA from gastric cancer patient's plasma samples. Thus ATP4A and ATP4B down-regulation involves DNA methylation and methylated ATP4B DNA in plasma is a potential biomarker for gastric cancer.