• The Journal of Korean Obstetrics and Gynecology
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• v.21 no.2
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• pp.38-48
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• 2008

Purpose: This study was conducted to investigate the effects of Hominis Placenta (紫河車) on the growth of human uterine myoma cells and cell apoptosis. Methods: Human uterine leiomyoma cells were cultured and treated with Hominis Placenta extract for 48 hours. Cell proliferation and activity was analyzed by MTT assay. We analyzed the cell cycle of human uterine myoma cells treated Hominis Placenta extract by FACS. Expression of proteins related to cell apoptosis (Bax, Bcl-2), cyclin-D1 and VEGF were evaluated by Western blotting method. Results: The human uterine myoma cells treated by Hominis Placenta extract didn't proliferate below the concentration of $10mg/m{\ell}$. And there was no remarkable difference on cell cycle analysis below the concentration of $10mg/m{\ell}$. The expression of Bax was decreased and the expression of Bcl-2 was increased after the treatment of Hominis Placenta extract. But the expressions of cyclin-D1 and VEGF were increased after the treatment of Hominis Placenta extract. Conclusion: This study suggests that Hominis Placenta induce uterine myoma cell apoptosis and have effect on the myoma cell proliferation in the concentraion below $10mg/m{\ell}$.

• Journal of Embryo Transfer
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• v.13 no.2
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• pp.159-172
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• 1998

사람 성장호르몬 수용체(hGHR) cDNA는 PCR방법에 의하여 fagment로서 보고되어진 바 있으나, liver cDNA로 부터 전장을 cloning한 보고는 없는 실정으로 본 연구에서는 기능을 가진 약 4.6kbp의 cDNA hGHR을 cloning 하는데 성공하였다. 먼저 cloning하기 위하여 human liver mRNA와 human breast cancer tissue로부터 회수한 mRNA를 RT-PCR방법에 의하여 human cDNA library와 cloning에 필요한 probe를 제작하였다. human library mRNA는 GT-PCR방법에 의하여 증폭하여 증폭되어진 산물은 λZAP Vector를 이용하여 cDNA library를 구축하였고,screeing을 위하여 임 보고 되어진 hGHR fragment native sequence를 기초로 N-terminal부분의 primer를 설계하여 950bp의 probe를 얻는데 성공하였다. 이 probe를 이용하여 준비된 human liver cDNA library로부터 2.5$\times$10 6개의 plaque로부터 6개의 positive clone을 획득하였고, 이들중 poly Asignal인 "AATAAA"를 포함하고 있는 가장 긴 약 3.8kbp의 clone을 sequencing한 결과 open reading frame을 포함하고 있었으나, 5'부분의 결손되어 있었다. 그리하여 이 부분은 human breast cancer tissue로 부터 회수한 mRNA를 RT-PCR에 의하여 증폭하였고, sequencing결과 이미 보고되어진 native hGHR와 비교한 결과 하나의 nucleotide가 silent mutation으로 판명되었다.한편 human liver cDNA library로부터 cloning한 3.8cp의 positive clone의 5'end의 결손된 부분에 silent mutation된 PCR 산물을 연결함으로써 native hGHR와 유사한 cDNA hGHR subcloning에 성공하였다. 이러한 cDNA hGHR의 clone이 function을 가지고 있는지를 검토하기 위하여 eukaryotic 발현 vector인 pCXN2에 의거 ligation한 후 chinese hamster ovary cell[CHO-KI]에 transfect를 실시하였다. Dexamethasone은 첨가하지 않고 hGH만의 존재하에서 이들 cell을 배양시키고 cell menbrane에서 발현 여부를 판정키 위하여 hGHR monocloual antibody를 사용하여 flow cytometery해석을 실시하는 한편 125I-hGH binding assay에 의하여 hGH binding activity를 측정하였다. 최종적으로 GH signal transduction의 target genedf으로 알려져 있는 serine protease inhibitor 2.1(Spi 2.1) gene의 promotor activity를 검토한 결과 hGHR을 transfect한 CHO Cell에 있어서 hGH의 농도에 의존적으로 증가되었다. 따라서 본 실험에서 cloning한 cDNA hGHR는 native hGHR와 같은 기능을 가지는 것으로 판명되었다.것으로 판명되었다.

• YAKHAK HOEJI
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• v.52 no.4
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• pp.252-258
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• 2008

The cytotoxic effect of benzoic acid and related compounds on the growth of normal cell lines and human oral epithelioid cell line was evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and 2,3-bis-[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-caboxanilide (XTT) methods. 3,4,5-Trihydroxybenzoic acid decreased the cell viability of human oral epithelioid cells and the cell adhesion activity of human oral epithelioid cells. Under the light microscopy, 100 ${\mu}M$ 3,4,5-trihydroxybenzoic acid showed the highest cytotoxic activity. From these results, we can propose that 3,4,5-trihydroxybenzoic acid has a potential anticancer activity.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.1
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• pp.91-96
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• 2015

A lipopeptide extract of Bacillus amyloliquefaciens BACY1 (BLE) was found to induce cell death in human oral squamous cell carcinoma (OSCC) cell lines, SCC4 and SCC25, in this study. The results of MTT assay showed that BLE inhibited OSCC cell proliferation in a dose-dependent manner. BLE was also effective in increasing the sub-G1 phases. Furthermore, when membrane damage in SCC4 cells treated with BLE was monitored by LDH assay, release of LDH was significantly increased. The protein and mRNA levels of pro-apoptotic Bax, and caspase-3 were up-regulated by BLE. Taken together, these results suggest that BLE induces apoptosis and then inhibits the cell proliferation of human OSCC cells.

• Biomedical Science Letters
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• v.11 no.3
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• pp.253-258
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• 2005

Reactive oxygen species (ROS) and sex hormones affect the proliferation of cells and are believed to play important roles in tumorigenesis. However, little is known regarding how these two factors interact to affect cell proliferation. In this study, hepal-6 cells were treated with ROS and sex hormones (testosterone and steroidal) either separately or in combination. The sex hormones had no significant influence the cell proliferation up to a concentration of $1{\mu}M$. However, cell proliferation was inhibited when the cells were treated simultaneously with $H_2O_2$, which alone was found to promote cell proliferation at the concentrations of $15{\mu}M$. In conclusion, this study indicates that instead of promoting the cell proliferation, ROS interact with sex hormones to inhibit the Hepa 1-6 cell proliferation.

• Journal of Nutrition and Health
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• v.31 no.4
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• pp.799-808
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• 1998

To investigate the antitumor activity of mugwort (Artemisia princeps Pampan), petroleum ether extract of mugwork was partially purified by a silica gel chromatography. Among several fractions, the fraction which was obtained under the elution with acetone, showed potent cytotoxicity against mouse leukemia cell line(Ll210), human colon cancer cell line (HCT-48) and human hepatoma cell line (Hep G2) , but was less effective with normal cell line(mouse embryo cell). Acetone fraction appeared to be glycolipid by Benedict test and the major fatty acids of the lipid were C16 ; 0 , C 18: 3by GC/MS analysis.

• The Journal of Korean Medicine
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• v.30 no.6
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• pp.17-26
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• 2009

Objectives: Little information is available regarding the effect of Lycii cortex radicis (LCR) on cell viability in glioma cells. This study was therefore undertaken to examine the effect of LCR on cell survival in U87MG human glioma cells. Methods: Cell viability and cell death were estimated by MTT assay and trypan blue exclusion assay, respectively. Reactive oxygen species (ROS) generation was measured using the fluorescence probe DCFH-DA. Activation of Akt and extracellular signal-regulated kinase (ERK) and activation of caspase-3 were estimated by Western blot analysis. Results: LCR resulted in apoptotic cell death in a dose- and time-dependent manner. LCR increased reactive oxygen species (ROS) generation and LCR-induced cell death was also prevented by antioxidants, suggesting that ROS generation played a critical role in LCR-induced cell death. Western blot analysis showed that LCR treatment caused down-regulation of Akt and ERK. The LCR-induced cell death was increased by the inhibitors of Akt and ERK. Activation of caspase-3 was stimulated by LCR and caspase inhibitors prevented the LCR-induced cell death. Conclusion: These findings suggest that LCR results in human glioma cell death through a mechanism involving ROS generation, down-regulation of Akt and ERK, and caspase activation.

• Toxicological Research
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• v.21 no.1
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• pp.77-85
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• 2005

Cadmium is an environmental pollutant exposed from contaminated foods or cigarette smoking and known to cause oxidative damage in organs. We investigated the cadmium-induced apoptosis and cell arrest in human breast cancer cells, MCF-7 cells and MDA-MB-231 cells. Obvious apoptotic cell death was shown in CdCl₂ 100 μM treatment for 12 hr, which were determined by DAPI staining and flow cytometric analysis. In cell cycle analysis, MCF-7 cells and MDA-MB-231 cells were arrested in S phase and G2/M phase respectively. These could be explained by the induction of cell cycle inhibitory protein, p21/sup Waf1/Cip1/ and p27/sup Kip1/, expression and reduction of cyclin/Cdk complexes in both cell lines. The decreased expression of cyclin A and Cdk2 in MCF-7 cells and cyclin B1 and Cdc2 in MDA-MB-231 cells were consistent with the flow cytometric observation. p-ERK expression was increased dose-dependent manner in both cell lines. It suggests that ERK MAPK pathway are involved in cadmium-induced cell cycle arrest and apoptosis. Moreover, cotreatment of zinc (100 μM, 12 hr) recovered the cadmium-induced cell arrest in both cells, which shows cadmium-induced oxidative stress mediates apoptosis and cell cycle arrest in human breast cancer cells.

• Reproductive and Developmental Biology
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• v.30 no.4
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• pp.287-291
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• 2006

We have developed a new passaging technique for the expansion of human embryonic stem cells (hESCs) that involves simply pipetting portions of hESCs acquired from colonies, reducing the laborious and time-consuming steps in the expansion of hESCs. Compared to general mechanical methods of passaging, our pipetting method allowed hESCs colonies to be broken into small fragments, which showed significantly higher attachment rates onto feeder cell layers. This technique produced three times the number of hESCs colonies than conventional mechanical methods. In addition, this pipetting method allowed us to distinguish differentiated hESCs from undifferentiated hESCs during hESCs colony pipetting. The hESCs cultured by pipetting method displayed normal human chromosomes for over 60 passages. According to RT-PCR and immunohistochemical analysis, the hESCs successfully maintained their undifferentiated state and pluripotency which was also confirmed by teratoma formation in viva Therefore, the pipetting method described in this study is a useful tool to efficiently and quickly expand hESCs on a large scale without enzyme treatment.

• Korean Journal of Plant Resources
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• v.30 no.6
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• pp.679-685
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• 2017

Stachys affinis tubers are known for its high content of stachyose and eaten as an edible vegetable. In this work, we assessed on the anti-inflammatory and anti-proliferation activity of a various type of extracts derived from S. affinis tubers. The n-hexane and dichloromethane fractions were showed the high cytotoxicity on the cell lines including RAW264.7 macrophages, HEK293 human kidney cell, A549 human lung cancer cell, KB human oral cancer cell, and a PC-3 human prostate cancer cell. N-butanol and water fractions were not exhibited cytotoxicity on the tested cancer cells, limited in anti-inflammatory and anti-cancer activities. Nevertheless, the ethyl acetate fraction showed little harm to RAW264.7 cells but inhibited the production of nitric oxide (NO) and prostaglandin E2 (PGE2) significantly. In addition, it arrests the cell growth in A549, KB, and PC-3 cell while little cytotoxicity on HEK293 cells. Consequently, these results supported that the ethyl acetate fraction of S. affinis tubers could be a potential anti-inflammatory and anti-cancer ingredient.