• 대한한의학회지
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• 제27권1호
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• pp.23-35
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• 2006

Objectives : This study was conducted to investigate the inhibitory effects of Gaejibokryunghwan on cell proliferation in HeLa cells. Methods : Human uterine cervical carcinoma HeLa cells were cultured in the 1%, 5% and 10% concentration of Gaejibokryunghwan extract solution. All three were cultured for 24 hours, 48 hours and 72 hours each, to examine the inhibitory effects of Gaejibokryunghwan. Afterwards, we drew out the effect of Gaejibokryunghwan extract solution by making 5 analysis. First analysis was to measure the proliferation rate of cells. Second was FACS analysis. Third was to estimate the activity or caspase-3. Fourth, we used XTT assay to analyze the activation or cells. Ana lastly, a molecular biological method was used to determine activation of MAP kinase in the HeLa cells. Results : After 24, 48 and 72 hours cultivation, the proliferation of HeLa cells showed the dose-dependent decrease in all Gaejibokryunghwan extract solution groups compared to the control group. In the FACS analysis, Gaejibokryunghwan extract solution groups showed increased caspase expression compared to the control group, except for the group for 48 and 72 hours in 1 % concentrate. Caspase-3 activities were increased in all, except tile group cultured for 24 hours in 5% concentrate and the groups cultured for 48 hours in 1% and 5% concentrate. In the XTT study, 1% Gaejibokryunghwan extract solution groups showed increase compared to the control group, but other Gaejibokryunghwan extract solution containing groups showed significant decrease compared to the control after 24, 48 and 72 hours of cultivation. The expressions of MAP kinase were decreased in all Gaejibokryunghwan extract solution containing groups compared to the control group after 24, 48 and 72 hours of cultivation. Conclusions : From this study, we could suggest that Gaejibokryunghwan be available to the inhibition of proliferation of human cervical carcinoma cell line, HeLa cells in vitro.

• Asian Pacific Journal of Cancer Prevention
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• 제13권5호
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• pp.2015-2020
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• 2012

Oleanolic acid (OA) is a naturally occurring triterpenoid in food materials and is a component of the leaves and roots of Olea europaea, Viscum album L., Aralia chinensis L. and more than 120 other plant species. There are several reports validating its antitumor activity against different cancer cells apart from its hepatoprotective activity. However, antitumor activity against skin cancer has not beed studied well thus far. Hence the present study of effects of OA against HaCaT (immortalized keratinocyte) cells - a cell-based epithelial model system for toxicity/ethnopharmacology-based studies - was conducted. Radical scavenging activity ($DPPH{\cdot}$) and FRAP were determined spectrophotometrically. Proliferation was assessed by XTT assay at 24, 48 and 72 hrs with exposure to various concentrations (12.5-200 ${\mu}M$) of OA. Apoptotic induction potential of OA was demonstrated using a cellular DNA fragmentation ELISA method. Morphological studies were also carried out to elucidate its antitumor potential. The results revealed that OA induces apoptosis by altering cellular morphology as well as DNA integrity in HaCaT cells in a dose-dependent manner, with comparatively low cytotoxicity. The moderate toxicity observed in HaCaT cells, with induction of apoptosis, possibly suggests greater involvement of programmed-cell death-mediated mechanisms. We conclude that OA has relatively low toxicity and has the potential to induce apoptosis in HaCaT cells and hence provides a substantial and sound scientific basis for further validation studies.

• 한국산업보건학회지
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• 제27권3호
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• pp.193-200
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• 2017

Objectives: This study was performed to evaluate the neurototoxicity of the environmental pollutant lead acetate(LA) and the protective effect of the D-2-amino-5-phosphonovaleric acid(APV), N-methyl-D-aspartate(NMDA) receptor antagonist on LA-induced cytotoxicity in cultured C6 glioma cells. Materials and Methods: For this study, cell viability in cultured C6 glioma cells was assessed by XTT assay and antioxidative effect, such as lactate dehydrogenase(LDH) activity, by LDH detection kit. Results: LA significantly decreased cell viability in a dose-dependent manner, and the XTT50 value was determined to be 33.3 uM of LA. The cytotoxicity of LA was deemed highly toxic according to Borenfreund and Puerner's toxic criteria. The vitamin E antioxidant significantly increased cell viability damaged by LA-induced cytotoxicity in these cultures. For the protective effect of APV on LA-induced cytotoxicity, APV significantly increased not only cell viability, but also inhibition of LDH activity. From these results, it is suggested that oxidative stress is involved in the neurotoxicity of LA, and APV effectively protected against LA-induced cytotoxicity via an antioxidative effect as an inhibotory activity of LDH. Conclusions: Natural resources like APV may be putative therapeutic agents for the toxic diminution of environmental pollutants such as LA correlated with oxidative stress.

• 동의생리병리학회지
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• 제17권2호
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• pp.503-509
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• 2003

The purpose of this study is to examine the toxic effects caused by xanthine oxidase/hypoxanthine (XO/HX) and the effects of herbal extracts such as Mangeum-tang (萬金湯: MGT) and Gamimangeum-tang (加味萬金湯: GMGT) on the treatment of the toxic effects. The results of these experiments were XO/HX, an oxygen radical-generating system, decreased the survival rates of the cultured cells on XTT assay, the amount of DNA syntheses, and the amount of neurofilaments, and increased c-fos positive cells, MGT and GMGT have the efficacy of increasing the survival rates of the cultured cells by increasing the amount of neurofilaments and DNA synthesis and decreasing the c-fos positive cells damaged by XO/HX, From the above results, it is suggested that MGT and GMGT have marked efficacy as a treatment for the damages caused by the XO/HX-mediated oxidative stress. And MGT and GMGT are thought to have certain pharmacological effects.

• 대한임상검사과학회지
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• 제43권3호
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• pp.98-104
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• 2011

To evaluate the physioactivity of Salicornia herbacea L. (SH), which are obtained from Sunchon bay as wild plants, an SH extract was prepared by freeze drying to obtain SH, and by cold drying to obtain SH. For the evaluation of their bioactivities, cell viability and antioxidative effect were measured. The XTT assay was adopted to measure cell viability after C6 glioma cells were treated with various concentrations of reactive oxygen species (ROS) and hydrogen peroxide ($H_2O_2$) for 8 hours. The DPPH-radical scavenging activity was also measured for the antioxidative effect. In this study, the $XTT_{50}$ value of $H_2O_2$ was determined at $30{\mu}M$ which was highly toxic based on the cytotoxic criteria by Borenfreund and Puerner. The protective effect of SH extract significantly increased cell viability compared with $H_2O_2$-treated group. Its antioxidative effect showed a significant DPPH-radical scavenging activity at concentrations of $1-100{\mu}g/mL$, while SH extract showed highly a DPPH-radical scavenging activity at only $100{\mu}g/mL$. From these results, $H_2O_2$ was highly toxic in cultured C6 glioma cells, and SH extract was effective in the prevention of cell damage by its antioxidative effect.

• Toxicological Research
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• 제11권2호
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• pp.175-180
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• 1995

We found that bufalln, one of the prominent components of the bufadlenolides in the Chinese medicine chan'su, has the potent inhibitory effects on growth and proliferation of the cultured bovine aortlc endothelial (BAE) and human umbilical vein endothelial (HUVE) cells. All naturally-occuring bufadienolides used in this study inhibited the cell growth in a dose-dependent manner. Particularly, bufalin among the bufadienolides showed the strongest inhibitory activity for the cell growth. The order of growth inhibition by bufadienolides on BAE cells was as follows: bufalin > gamabufotalln > bufotalln > cinobufagin > cinobufotalin > resibufogenin. The $IC_50$ values (50% inhibition of cell growth) of bufalin as determined by XTT assay were the range of 1-10 nM in BAE and HUVE cells. Bufalin exhibited a higher sensitivity towards cultured bovine aortic endothelial cells than human umbilical vein endothelial cells.

• Natural Product Sciences
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• 제5권4호
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• pp.162-164
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• 1999

Dehydroaltenusin (1) was isolated from the chloroform extract of the culture filtrate of a Streptomyces sp. and its structure was determined from spectral data as well as by comparison with published values. In an XTT-based in vitro anti-HIV assay, dehydroaltenusin effectively inhibited the cytopathic effects of HIV infection at a concentration of $1-5\;{\mu}g/mL$.

• 약학회지
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• 제45권6호
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• pp.617-622
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• 2001

A protein-polysaccharide fraction of a Korean wild mushroom Psathyrella velutina, PVP, was prepared and its antitumor immunomodulatory activity was investigated. When PVP was administered once daily for seven days from day 1 to day 7 into male ICR mice implanted with 1 $\times$ 10$^{5}$ cells of sarcom 180 tumor cells into the peritoneum on day 4, it inhibited the growth of sarcoma 180 cells by 92.8%. In XTT assay, PVP also exhorted in vitro anti-proliferation activity on U-937, a human monoblastoid cell line, as well as sarcoma 180 cells. PVP showed marked stimulatory activity on the immune system in that it induced the accumulation of PEC (the stimulation index, Sl=4.90 at 100 mg/kg), stimulated the BALB/c mouse splenic lymphocytes to form lymphoblasts (Sl=5.75 at 100$\mu\textrm{g}$/ml), and upergulated the expression of CD25 molecules. All these results strongly support that PVP exhorts its antitumor activity through stimulation of the immune system as well as anti-proliferative activity on the tumor cells.

• 한국식품영양과학회지
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• 제31권4호
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• pp.691-695
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• 2002

한국산 솔잎을 가압.압착하여 얻은 수액을 증류한 솔잎 수액 증류액의 각종 암세포주에 대한 in uitro 세포독성을 시료액 대비 10배, 20배, 40배 희석군과 대조군에 대해 XTT법으로 실험한 결과, 쥐 백혈병 세포주인 L1210에 대해서는 76~89%, 쥐 육종암세포인 sarcoma 180에 대해서는 61~90%의 세포성장 억제효과를 나타내었다. 또한 인체의 monocyte-like cancer cell인 U937에 대해서는 56~81%, 인체 유방암 세포주인 T47D와 MDA-MB-231에서는 각기 12%, 또 다른 유방암 세포주인 MH7A에서는 64%, 인체 간암 세포주인 SNU-354에 대해서는 72%의 높은 세포 증식 억제효과를 나타내었다. 이로써 본 연구 시료인 솔잎 수액 증류액은 쥐 백혈병 세포주인 L1210, 쥐 육종암세포인 sarcoma 180, 인체 monocyte-like cancer cell인 U937, 인체 유방암 세포주인 MH7A, 인체 간암 세포주인 SNU-354에 대해 강한 세포독성을 갖는 것을 알 수 있었다. 이와 함께 솔잎 수액 증류액의 암세포에 대한 독성효과는 솔잎의 처리과정에 따라 다를 수 있으며, 또한 동일한 솔잎 수액증류액의 농도에서도 암세포주 종류에 따라 세포독성정도가 다름을 알 수 있었다. 따라서 최적 투여 농도와 적용 암세포주를 찾을 경우 새로운 항암제로 개발될 수 있음을 제시하였다.

• 대한한방부인과학회지
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• 제19권1호
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• pp.14-30
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• 2006

Purpose : This study was conducted to investigate the inhibitory effects of Cortex ulmi pumilae on cell proliferation in HeLa cell. Methods : Human uterine cervical carcinoma HeLa cells were cultured in the 1%, 5% and 10% concentration of Cortex ulmi pumilae solution for 24 hours, 48 hours and 72 hours for the direct inhibitory effects of Cortex ulmi pumilae. Afterwards, we executed the analysis of the effect of Cortex ulmi pumilae solution on cell proliferation inhibition using XTT assay, DNA fragmentation, molecular biological method through MAP kinase activity and FACS analysis of caspase activity in the HeLa cells. Results : After 48 and 72 hours cultivation, the HeLa cells showed the concentration-dependently significant increase in all Cortex ulmi pumilae solution containing groups compared to the control. In the FACS analysis, all Cortex ulmi pumilae solution containing groups showed concentration-dependent increase compared to the control after 24 hours cultivation and the caspase-3 activities were decreased in all Cortex ulmi pumilae solution containing groups compared to the control after 24, 48 and 72 hours cultivation. After 48 and 72 hours cultivation, we could examined the apparent DNA fragmentation in all Cortex ulmi pumilae solution containing groups. In the XTT study, all Cortex ulmi pumilae solution containing groups showed concentration-dependent decrease compared to the control after 24 and 72 hours cultivation but 10% group after 48 hours and 5% and 10% groups after 72hours were presumed statistically significant differences. The expressions of MAP kinase were decreased in all Cortex ulmi pumilae solution containing groups compared to the control after 24, 48 and 72 hours cultivation. Conclusion : From this study we could suggest that Cortex ulmi pumilae be available to the inhibition of apoptosis of human cervical carcinoma cell line in vitro.