• Applied Biological Chemistry
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• v.50 no.3
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• pp.228-232
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• 2007

This study was performed to investigate the cytotoxic activity from Vitex rotundifolia. V. rotundifolia was extracted with methanol, ethanol, and acetone, and then the cytotoxic effect of these extracts was measured by the MTT reduction assay and morphological assay on the HT-29 human colon carcinoma cells. Among the three extracts, the acetone extract showed the highest cytotoxic activity on the HT-29 cells in a dose-dependent manner with an $IC_{50}$ value of 10 ${\mu}g/ml$. The acetone extract was further fractionated sequentially with n-hexane, diethyl ether, ethyl acetate, and water layer according to the degree of polarity. The n-hexane layer among the fractioned layers showed inhibitory activity on the growth of HT-29 cells. In addition, n-hexane layer also showed the cytotoxic activity against SW620 human colon carcinoma cells. These result indicated that extracts of V. rotundifolia may contain bioactive materials and could be potential candidates as chemotherapeutic agents against human colon carcinoma cells.

• Applied Biological Chemistry
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• v.49 no.3
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• pp.248-253
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• 2006

The present study describes the preliminary evaluation of the cytotoxicity from Gleditsiae Semen extracts. G. Semen was extracted with methanol, ethanol, and acetone, and then cytotoxic effect of these extracts was measured by the MTT reduction assay and phase-contrast microscopy on the HT-29 human colon carcinoma cells. Among these extracts, methanol extract showed the highest cytotoxic activity on the HT-29 cells. The methanol extract was further fractionated with n-hexane, diethyl ether, ethyl acetate, and water layer according to the degree of polarity. The water layer showed the highest inhibitory activity on the growth of HT-29 cells, but the other fractions indicated the low cytotoxic activity. In addition, water layer also showed the cytotoxic activity against SW620 human colon carcinoma cells. Based on these results, we suggest that extracts of G. Semen may contain bioactive materials and are potential candidates as chemotherapeutic agents against human colon carcinoma cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.9
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• pp.1099-1105
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• 2007

The present study describes the preliminary evaluation of the anticancer activity of Styela plicata. Freeze-dried S. plicata was extracted with methanol, ethanol, acetone, and water, and then anticancer effect of the extracts was measured by the MTT reduction assay and phase-contrast microscopy on the HT-29 human colon carcinoma cells. Among the extracts, acetone extract showed the highest anticancer activity. The cell proliferation rates markedly decreased by 94.0% at the concentration of 500 ${\mu}g/mL$ of acetone extract compared with control cells. The acetone extract was further fractionated with hexane, diethyl ether, ethyl acetate, and water layer according to the degree of polarity. The HT-29 cells with hexane layer extract (250 ${\mu}g/mL$) decreased the cell viability to 5.1% of untreated control. The growth of SW620, HeLa, and MCF-7 cells was decreased to about 10%, by the treatment of hexane layer extract 250 ${\mu}g/mL$. Theses results suggest extracts from S. plicata as possible natural cancer therapeutic material.

• Radiation Oncology Journal
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• v.19 no.2
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• pp.163-170
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• 2001

Purpose : The measurement of radiation survival using a clonogenic assay, the established standard, can be difficult and time consuming. In this study, We have used the MTT assay, based on the reduction of a tetrazolium salt to a purple formazan precipitate by living cells, as a substitution for clonogenic assay and have examined the optimal condition for performing this assay in determination of radiation sensitivity. Materials and Methods : Four human cancer cell lines - PCI-1, SNU-1066, NCI-H630 and RKO cells have been used. For each cell line, a clonogenic assay and a MTT assay using Premix WST-1 solution, which is one of the tetrazolium salts and does not require washing or solubilization of the precipitate were carried out after irradiation of 0, 2, 4, 6, 8, 10 Gy. For clonogenic assay, cells in $25\;cm^2$ flasks were irradiated after overnight incubation and the resultant colonies containing more than 50 cells were scored after culturing the cells for $10\~14$ days. For MTT assay, the relationship between absorbance and cell number, optimal seeding cell number, and optimal timing of assay was determined. Then, MTT assay was performed when the irradiated cells had regained exponential growth or when the non-irradiated cells had undergone four or more doubling times. Results : There was minimal variation in the values gained from these two methods with the standard deviation generally less than $5\%$, and there were no statistically significant differences between two methods according to t-test in low radiation dose (below 6 Gy). The regression analyses showed high linear correlation with the $R^2$ value of $0.975\~0.992$ between data from the two different methods. The optimal cell numbers for MTT assay were found to be dependent on plating efficiency of used cell line. Less than 300 cells/well were appropriate for cells with high plating efficiency (more than $30\%$). For cells with low plating efficiency (less than $30\%$), 500 cells/well or more were appropriate for assay. The optimal time for MTT assay was after 6 doubling times for the results compatible with those of clonogenic assay, at least after 4 doubling times was required for valid results. In consideration of practical limits of assay (12 days, in this study) cells with doubling time more than 3 days were inappropriate for application. Conclusion : In conclusion, it is found that MTT assay can successfully replace clonogenic assay of tested cancer cell lines after irradiation only if MTT assay was undertaken with optimal assay conditions that included plating efficiency of each cell line and doubling time at least.

• The Journal of Korean Medicine
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• v.20 no.1 s.37
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• pp.91-105
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• 1999

The effects of Yinjin and Yinjinsaryongsangagambang on a DNA damaging agent, etoposide-induced apoptosis, cell viability, cell cycle progression, and mRNA expression of apoptosis-related genes of human hepatocyte cell line HepG2 were investigated using tryphan blue exclusion assay, MTT assay, flow cytometry, immunocytometric analysis of PCNA, and quantitative RT-PCR analysis. MTT assay showed that Yinjin and Yinjinsaryongsangagambang increases cellular viability of HepG2 cells in a dosage-dependent manner. Stimulation of cell cycle progression by Yinjin or Yinjinsaryongsangagambang was detected by flow cytometric analysis of the DNA content and immunocytometric analysis of PCNA expression. A significant reduction of a DNA-damaging agent, etoposide-induced apoptosis were found in both Yinjin and Yinjinsaryongsangagambang-treated cells in dosage-dependent manner. In overall, 3-fold reduction of apoptosis was recognized in $10.0\;{\mu}g/ml$ of Yinjin or Yinjinsaryongsangagambang-treated cells compared to untreated cells. Although the difference is not significant, Yinjinsaryongsangagambang showed slightly higher effect on the inhibition of apoptosis than Yinjin. From flow cytometric analysis of apoptosis, while 39.9% of untreated cells showed etoposide-induced apoptotic cell death, only 19.6% or 17.4% of Yinjin or Yinjinsaryongsangagambang-treated cells were fond at apoptotic sub G1 phase, respectively. Interestingly, strong induction of Gadd45-mRNA was observed from Yinjin or Yinjinsaryongsangagambang-treated cells. However, no changes in expression levels of p53 and Waf1 were detected, demonstrating that induction of Gadd45 mRNA expression by Yinjin or Yinjinsaryongsangagambang occurs by p53-independent mechanism. Marked mRNA inductions of two apoptosis-inhibiting genes, Bcl-2 and Bcl- XL, were found in both Yinjin or Yinjinsaryongsangagambang-treated HepG2 cells while no changes was detected in expression levels of an apoptosis-promoting gene, Bax.

• Journal of Environmental Health Sciences
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• v.23 no.2
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• pp.98-105
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• 1997

The study on the cytotoxicity of heavy metals was carried out to evaluate the cytotoxic effect of those on mouse L929 fibroblast cell in 96-well microtiter plates. The cytotoxicity was assayed by the neutral red, tetrazolium MTT, total protein, micronuclei test. The cytotoxicity of the heavy metals by neutral red and tetrazolium MTT was showed in order, cadmium > zinc > nickel for the cationic metals tested. The effect of metal-metal interaction on the cytotoxicity showed a marked reduction of cadmium toxicity by zinc, to a lesser degree, by nickel. The amount of total protein in treated group added heavy metals was less than that of the control and treated cadmium alone was less than those of combination with nickel or zinc. At midpoint cytotoxicity values of heavy metals, the frequency of micronuclei on the cell treated heavy metals was more than that of control and treated cadmium alone was more than those of combination with nickel or zinc. From those results, it could be suggested that the heavy metals decreased the viability of mouse fibroblast L929 cells in a concentration-dependent manner and have cytogenic toxic effects, but mixed group decreased the cytotoxic and cytogenic toxicity on L929 cells.

• Journal of Plant Biotechnology
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• v.3 no.2
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• pp.83-88
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• 2001

A cell culture system of poplar (Populus alba x P.glandulosa) was established to test four different methods for evaluation of cellular stresses. Two different kinds of stresses were given to the cultures by adding either Pb(NO$_3$)$_2$ or paraquat and the cellular responses were monitored during a week period. While fresh weight reduction was observable in two days after the treatment of Pb(NO$_3$)$_2$, such changes were apparent only in later stage in paraquat treated cultures. Cells in paraquat treated cultures in the first 3 days showed no alteration in fresh weight as compared to untreated cultures, but had their MTT reducing activities completely inhibited. Neither Evans blue staining nor ion conductivity of the medium was consistent with fresh weight changes of the cultures. Overall, cell clumps formed during suspension culture appeared to interfere with staining and washing reactions and thus cause the assays unreliable. Among the four methods examined, fresh weight changes and MTT reducing activity appeared to be the most reliable and consistent.

• Applied Biological Chemistry
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• v.50 no.1
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• pp.63-67
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• 2007

The neuroprotective effect of methanolic extracts from Dendrobium nobile Lindl. (DME) against $H_{2}O_{2}$-induced neurotoxicity in PC12 cells was investigated. The treatment of PC12 cells with various DME concentrations under $H_{2}O_{2}$ resulted in the induction of protective effect in a dose-dependent manner, as determined by the results of an MTT reduction assay, an LDH release assays, and a morphological assay. Interestingly, we also detected reduction of apoptotic bodies and inhibition of caspase-3 activity by DME in $H_{2}O_{2}$-indeced PC12 cells. These data show that the neuroprotective effect of DME against PC12 cells might be related to the suppression of caspase-3 activation. Therefore, these results suggest that DME could be a new potential candidate as chemotherapeutic agents against neuronal diseases.

• Journal of radiological science and technology
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• v.37 no.1
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• pp.21-28
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• 2014

Cultured pancreatic beta cells and nerve cells, it is given normal condition of 10% FBS (fetal bovine serum), 11.1 mM glucose and hyperglycemia codition of 1% FBS, 30 mM glucose. For low LET X-ray irradiated with 0.5 Gy/hr dose-rate(total dose: 0.5 to 5 Gy). Survival rates were measured by MTT assay. When non irradiated, differentiated in the pancreatic beta cells experiment is hyperglycemia conditions survival rate compared to normal conditions survival rate seemed a small reduction. However increasing the total dose of X-ray, the survival rate of normal conditions decreased slightly compared to the survival rate of hyperglycemia conditions, the synergistic effect was drastically reduced. When non irradiated, undifferentiated in the nerve cells experiment is hyperglycemia conditions survival rate compared to normal conditions survival rate seemed a large reduction. As the cumulative dose of X-ray normal conditions and hyperglycemia were all relatively rapid cell death. But the rate of decreased survivals by almost parallel to the reduction proceed and it didn't show synergistic effect.

• Journal of Life Science
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• v.29 no.11
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• pp.1192-1199
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• 2019

The intention of this study was to confirm the possible use of an ethanol extracts of Nelumbinis Rhizomatis Nodus (NRN) as a cosmetic material. To this end, we extracted NRN with 70% ethanol and performed biological activity evaluation of whitening efficacy and wrinkle reduction. We performed cellular tyrosinase inhibition and melanin contents assay to check the whitening activity of NRN and carried out a toxicity evaluation of NRN via an MTT assay and the amounts of associated proteins that affect melanin production in a melanoma cell line (B16F10). And collagenase inhibitory assay was performed for the evaluation of anti-wrinkle of samples. In addition, a toxicity evaluation using an MTT assay and matrix metalloprotease (MMP-1) and procollagen synthesis inhibition by NRN were evaluated in a fibroblast cell line (CCD-986sk). Western blot results for the whitening activity evaluation revealed that the levels of two proteins related to melanin production, tyrosinase-related protein-1 (TRP-1) and tyrosinase-related protein-2 (TRP-2), were decreased in a dose-dependent manner. Moreover, collagenase inhibition activity at a concentration of $500{\mu}g/ml$ NRN by measuring epigallocatechin-3-gallate (EGCG) was increased by more than 80% compared to the control group. Meanwhile, procollagen synthesis was reduced by 68.8% in the UVB-induced CCD- 986sk cells group whereas collagen synthesis recovered by 80.2% with $25{\mu}g/ml$ NRN. The MMP-1 expression rate showed 20.2% reduction at $25{\mu}g/ml$. The results of the experiments verified the whitening and wrinkle suppression effects of NRN and confirmed that it could be used as a safe natural cosmetic material in the future.