• Title/Summary/Keyword: MTT assay

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Effects of Apoptosis of Sophorae Radix on Human Gastric Adenocarcinoma cells (인체 위암세포에서 고삼의 세포사멸효과)

  • Lim, Bo-Ra;Lee, Hee-Jung;Kim, Min-Chul;Kim, Hyung-Woo;Kim, Byung-Joo
    • Korean Journal of Oriental Medicine
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    • v.18 no.1
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    • pp.85-92
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    • 2012
  • Objective : The purpose of this study was to investigate the anti-cancer effects of Sophorae Radix and the effects of 5-Fluorouracil (5-FU) in human gastric adenocarcinoma cells (AGS). Method : We used human gastric adenocarcinoma cell line, AGS cells. We examined cell death by MTT assay and caspase 3 assay with Sophorae Radix. To examine the inhibitory effects of Sophorae Radix, cell cycle (sub G1) analysis was done the AGS cells after three days with Sophorae Radix. The reversibility of Sophorae Radix was examined on one day to five days treatment with 100 ${\mu}g/ml$ Sophorae Radix. Result : Sophorae Radix inhibited the growth of AGS cells in a dose-dependent fashion. Also we showed that Sophorae Radix induced apoptosis in AGS cells by MTT assay, caspase 3 assay and sub-G1 analysis. Sophorae Radix combined with 5-FU markedly inhibited the growth of AGS cells compared to Sophorae Radix or 5-FU alone. After 3 days treatment of AGS cells with Sophorae Radix, the fraction of cells in sub-G1 phase was much higher than that of the control group. Conclusion : Our findings provide insight into unraveling the effects of Sophorae Radix in human gastric adenocarcinoma cells and developing therapeutic agents against gastric cancer.

Effects of Sophorae Radix on Human Breast Adenocarcinoma Cells (고삼의 인체 유방암세포에 미치는 효과)

  • Lee, Hee-Jung;Kim, Min-Chul;Lim, Bo-Ra;Bae, Go-Eun;Kim, Hyung-Woo;Kwon, Young-Kyu;Kim, Byung-Joo
    • Korean Journal of Oriental Medicine
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    • v.18 no.1
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    • pp.75-84
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    • 2012
  • Objective : The purpose of this study was to investigate the anti-cancer effects of Sophorae Radix and the effects of Doxorubicin (DOX) in human breast adenocarcinoma cells (MCF-7). Method : We used human breast adenocarcinoma cell line, MCF-7 cells. We examined cell death by MTT assay and caspase 3 assay with Sophorae Radix. To examine the inhibitory effects of Sophorae Radix, cell cycle analysis was done the MCF-7 cells after three days with Sophorae Radix. The reversibility of Sophorae Radix was examined on one day to five days treatment with 100 ${\mu}g/ml$ Sophorae Radix. Result : Sophorae Radix inhibited the growth of MCF-7 cells in a dose-dependent fashion. Also we showed that Sophorae Radix induced apoptosis in MCF-7 cells by MTT assay, caspase 3 assay and sub-G1 analysis. Sophorae Radix combined with DOX markedly inhibited the growth of MCF-7 cells compared to Sophorae Radix or DOX alone. After 3 days treatment of MCF-7 cells with Sophorae Radix, the fraction of cells in sub-G1 phase was much higher than that of the control group. Conclusion : Our findings provide insight into unraveling the effects of Sophorae Radix in human breast adenocarcinoma cells and developing therapeutic agents against breast cancer.

Effects of Schisandra Chinensis on Human Breast Cancer Cells (오미자의 유방암 세포사멸과 TRPM7 관련성에 관한 연구)

  • Kim, Jung Nam;Chae, Han;Kwon, Young Kyu;Kim, Byung Joo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.2
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    • pp.162-168
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    • 2014
  • Fruits of Schisandra chinensis (SC) Baill are considered a traditional herbal medicine for the treatment and alleviation of various diseases. The purpose of this study was to investigate the anti-cancer effects of SC extract in human breast adenocarcinoma cells (MCF-7). We used human breast adenocarcinoma cell line, MCF-7 cells. We examined cell death by MTT assay and caspase 3 and 9 assay with SC extract. To examine the inhibitory effects of SC extract, cell cycle (sub G1) analysis and mitochondrial membrane depolarization was done the MCF-7 cells after one day with SC extract. In addition, to investigate the transient receptor potential melastatin 7 (TRPM7) currents, we used the whole cell patch clamp techniques. Furthermore, TRPM7 channels were overexpressed in human embryonic kidney (HEK) 293 cells to identify the role of TRPM7 channels in MCF-7 cell growth and survival. SC extract inhibited the growth of MCF-7 cells in a dose-dependent fashion. Also we showed that SC extract induced apoptosis in MCF-7 cells by MTT assay, caspase 3 and 9 assay, sub-G1 analysis and mitochondrial membrane depolarization. SC extract inhibited the TRPM7 currents in MCF-7 cells and in TRPM7 overexpressed HEK 293 cells. Furthermore, TRPM7 channel overexpression in HEK 293 cells exacerbated SC extract-induced cell death. Our findings provide insight into unraveling the effects of SC extract in human breast adenocarcinoma cells and developing therapeutic agents against breast cancer.

Antioxidant Activity and Neuroprotective Effect of Psoralea corylifolia Linne Extracts (파고지 (Psoralea corylifolia Linne) 추출물의 항산화 작용에 의한 신경세포 보호효과)

  • Yoon, Mi-Young;Lee, Bo-Bae;Kim, Ju-Young;Kim, Yong-Seong;Park, Eun-Ju;Lee, Seung-Cheol;Park, Hae-Ryong
    • Korean Journal of Pharmacognosy
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    • v.38 no.1
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    • pp.84-89
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    • 2007
  • The present study describes the preliminary evaluation of the antioxidant activities and the neuroprotective effect of methanolic extracts from Psoralea corylifolia Linne (PCE). The antioxidant activities and neuroprotective effect of the PCE were evaluated by total phenolic contents (TPC), DPPH radical scavenging activity (RSA), reducing power (RP), MTT reduction assay, and LDH release assay. TPC, DPPH RSA, and RP of the extract at concentration of 100 ${\mu}g$ was 125.93 ${\mu}g$, 63.81%, 0.138, respectively, and those were concentration dependent. The treatment of PC12 and N18-RE-105 cells with various PCE concentrations under $H_2O_2$ resulted in the induction of protective effect in a dose-dependent manner, as determined by the results of an MTT reduction assay and LDH release assay. Therefore, these results suggest that PCE could be a new potential candidate as an antioxidant against neuronal diseases.

Experimental Studies on the Change of Cytotoxic and Antitumor Effects according to the Prebrewed Method of Semen Tiglii and Rhizoma Coptidis (수치파두(修治巴豆) 및 파두가황연(巴豆加黃連)의 세포독성(細胞毒性)과 항종양(抗腫瘍) 효과(效果)에 관(關)한 실험적(實驗的) 연구(硏究))

  • Jo, Soung-Gak;Moon, Goo;Moon, Suk-Jae
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.1 no.1
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    • pp.191-211
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    • 1995
  • This experiment was designed to study the change of cytotoxic and antitumor effects according to the prebrewed method of Semen Tiglii and Rhizoma Coptidis. The cytotoxic and antitumor effects were evaluated on human cell lines(A 549, Caki-1, LL2, Sarcoma 180, NIH/3T3) after exposure to prebrewed Semen Tiglii and Rhizoma Coptidis water extract 0.1, 0.2, 0.4, 0.8, 1.6 mg/ml using in MTT assay, LDH, colony forming efficency and SRB assay which were regarded as a valuable method for cytotoxic and antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows. 1. The cytotoxicity from the result of MTT assay was low slightly in the ST II(炒巴豆霜), high in the ST III(醋炒巴豆). The cytotoxicity of ST I + RC(生巴豆霜加黃連) was similar to that of STI(生巴豆霜). 2. The cytotoxicity from the result of LDH was low slightly in the ST Ⅱ (炒巴豆霜), high in the ST III(醋炒巴豆). The cytotoxicity of ST I + RC(生巴豆霜加黃連) was similar to that of ST I(生巴豆霜). 3. The antitumor affect on A 549 tumor cell from the result of colony forming efficiency was low slightly in the ST II (炒巴豆霜) and ST I + RC(生巴豆霜加黃連). 4. The antitumor effect on Caki-1 tumor cell from the result of SRB assay was low slightly in the ST II (炒巴豆霜). 5. Median survival time and Increased life span increased slightly in the ST I RC(生巴豆霜加黃連) and ST II (炒巴豆霜). 6. The inhibitory effect on the growth of Sarcoma 180 and Lewis lung carcinoma tumor cell increased slightly in the ST I + RC(生巴豆霜加黃連) and ST II (炒巴豆霜).

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Analysis of Antibacterial Activity against Food Spoilage and Food-borne Pathogens and Cytotoxicity on Human Cancer Cell Lines of Extracts from Pericarp and Seed of Vitis coignetiea (머루 과피와 종자 추출물의 식품 위해성 세균에 대한 항균성 및 인체 암세포주에 대한 cytotoxicity 분석)

  • Won, Ji-Hye;Kim, Mee-Ra
    • Korean journal of food and cookery science
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    • v.28 no.2
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    • pp.175-182
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    • 2012
  • In this study, antibacterial activity and cytotoxicity of the extracts from pericarp and seed of $Vitis$ $coignetiea$, which were extracted with 0.1% HCl-60% ethanol, were analyzed. The antibacterial activity of the extracts was determined by paper disc diffusion method against food spoilage and food-borne pathogens. The pericarp extract showed high antibacterial activity against $Bacillus$ $cereus$, $Escherichia$ $coli$ O157:H7, and $Pseudomonas$ $aeruginosa$, and the seed extract represented the high antibacterial activity against $B.$ $cereus$, $E.$ $coli$ O157:H7, and $Staphylococcus$ $aureus$. The cytotoxicity of the $Vitis$ $coignetiea$ extract against human cancer cells was determined using the MTT assay and SRB assay. The pericarp extract represented strong growth-inhibition activity against G361 and Hep3B cells and the seed extract greatly inhibited the growth of HeLa and G361 cells in the MTT assay. In addition, the pericarp extract displayed a high inhibition activity against the growth of AGS cells and the seed extract greatly inhibited the growth of HeLa, Hep3B, and MCF7 cells in the SRB assay. Especially, the cytotoxicities of the seed extract against HeLa were significantly higher than those of the extract against other cancer cells at all test concentrations. This study demonstrates that the extract from pericarp and seed of $Vitis$ $coignetiea$ possess high antibacterial activity and cytotoxicity.

Neuroprotective Effects of Extracts from Diospyros kaki L. Peel (단감(Diospyros Kaki L.)껍질 추출물의 신경세포 보호 효과)

  • Lee, Mi-Ra;Moon, Seong-Hee;Choi, Ae-Ran;Lee, Seung-Cheol;Ahn, Kwnag-Hwan;Park, Hae-Ryong
    • Korean journal of food and cookery science
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    • v.27 no.4
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    • pp.67-73
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    • 2011
  • This study was performed to assess the neuroprotective effects of methanolic extracts from sweet persimmon peel (PPE) against glutamate-induced cytotoxicity in hybridoma N18-RE-105 cells. The neuroprotective effects of PPE in N18-RE-105 cells were measured using the MTT reduction assay, LDH release assay, and phase-contrast microscopy. The results of the MTT reduction assay showed that treating cells with 500 ${\mu}g/ml$ PPE resulted in cell viability of 66.9%. Additionally, the morphological changes and the results of the LDH release assay showed that glutamate-induced damage to nerve cells was strongly inhibited by PPE. GSH content of N18-RE-105 cells was 3.5 ${\mu}M$ compared to that of the control, whereas pretreatment with 500 ${\mu}g/ml$ PPE increased GSH content by 4.7 ${\mu}M$. PPE was fractionated with hexane, and that layer had the highest neuroprotective effects in glutamate-stressed N18-RE-105 cells. In conclusion, our data showed that glutamate potentiated the effects of N18-RE-105 cell death by a mechanism involving oxidative stress. Therefore, PPE may be a potential candidate for prevention and therapy of neurodegenerative diseases.

Inhibitory Effect of D-chiro-inositol on Both Growth and Recurrence of Breast Tumor from MDA-MB-231 Cancer Cells

  • Kim, Yoon-seob;Park, Ji-sung;Kim, Minji;Hwang, Bang Yeon;Lee, Chong-kil;Song, Sukgil
    • Natural Product Sciences
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    • v.23 no.1
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    • pp.35-39
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    • 2017
  • D-chiro-inositol (DCI) is a secondary messenger in insulin signal transduction. It is produced in vivo from myo-inositol via action of epimerase. In this study, we evaluated antitumor activity of DCI against human breast cancer both in vitro and in vivo. In order to determine the inhibitory effects of DCI on growth of human breast cancer cells (MDA-MB-231), two different assessment methods were implemented: MTT assay and mouse xenograft assay. MTT assay demonstrated downturn in cell proliferation by DCI treatment (1, 5, 10, 20 and 40 mM) groups by 18.3% (p < 0.05), 17.2% (p < 0.05), 17.5% (p < 0.05), 18.4% (p < 0.05), and 24.9% (p < 0.01), respectively. Also, inhibition of tumor growth was investigated in mouse xenograft model. DCI was administered orally at the dose of 500 mg/kg and 1000 mg/kg body weight to treat nude mouse for 45 consecutive days. On the 45th day, tumor growth of DCI (500 mg/kg and 1000 mg/kg) groups was suppressed by 22.1% and 67.6% as mean tumor volumes were $9313.8{\pm}474.1mm^3$ and $3879.1{\pm}1044.1mm^3$, respectively. Furthermore, breast cancer stem cell (CSC) phenotype ($CD44^+/C24^-$) was measured using flow cytometry. On the 46th day, CSC ratios of DCI (500 mg/kg) and co-treatment with doxorubicin (4 mg/kg) and DCI (500 mg/kg) group decreased by 24.7% and 53.9% (p < 0.01), respectively. Finally, from tumor recurrence assay, delay of 5 days in the co-treatment group compared to doxorubicin (4 mg/kg) alone group was observed. Based on these findings, we propose that DCI holds potential as an anti-cancer drug for treatment of breast cancer.

Effect of Bunsimgieumgagambang on the Stress Due to the Maternal Separation in Rats (분심기음가감방(分心氣飮加減方)이 모성분리(母性分離) stress 백서(白鼠)에 미치는 영향)

  • Kim Ki-Bong;Kim Jang-Hyun;Chang Gyu-Tae
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.5
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    • pp.1303-1310
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    • 2005
  • This study was performed to investigate the effect of Bunsimgieumgagam on the stress due to the maternal separation in rat. In this study, we researched in 'the behavioral observation', 'the changes of body weight', 'quantitative analysis of the number of BrdU-positive cells per section in dentate gyrus of hippocampus', 'free radical scavenging assay' and 'MTT-based cytotoxicity assay of SK-N-SH cell line', in order to figure out the effect on which Bunsimgieumgagam has the increase of neuron in dentate gyrus of hippocampus damaged by the stress due to the maternal separation. In the behavioral Observation, Bunsimgieumgagam was also efficacious against the decline of one's behavior and anorexia derived from the stress by the maternal separation. In the change of body weight, it showed that the Bunsimgieumgagam is effective in the recovery of weight loss caused by heavy stress(p<0.05). Also, Bunsimgieumgagam had an increasing effect, which is similar to a normal state, on DG's neuron in hippocampus (P<0.001). In free radical scavenging assay, Bunsimgieumgagam had a superior free radical scavenging effect. And it showed a significant result with the high cell proliferation effect in MTT-based cytotoxicity assay(P<0.01, p<0.001) This result suggest that Bunsimgieumgagam has an anti-stress effect and a proliferation effect of neuron in dentate gyrus of hippocampus, and it shows the potential of Bunsimgieumgagam in the treatment for the various disorders derived from children's stress.

Oxidative Stress in the Cell and Antioxidant Activity of Kalopanax Pictus Extracts (음나무 추출물의 세포 내 산화 스트레스와 항산화 활성)

  • Kim, Sea-Hyun;Park, Youngki;Jang, Yong-Seok;Han, Jingyu;Chung, Hun-Gwan
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.6
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    • pp.126-134
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    • 2007
  • This study reviewed the application of an extract from Kalopanax pictus stem bark and root bark as natural antioxidants. To investigate the effect on cell toxic level against transformed mouse fibroblast L929 in formula added with various extracts from Kalopanax pictus stem bark and root bark, this experiment was carried out by in vitro cytotoxicity method. Using DCFA-DA method, oxidative stress in cell was measured with other antioxidant activity methods including DPPH assay and NBT assay. Active oxygen inhibition rate for root bark insoluble hot water extracts showed the highest with 57.9% for 15 min treatment. In DPPH and NBT test, antioxidant activities of methanol extract from stem bark and insoluble hot water extract from stem bark were 96% (at 0.1%) and 95% (at 0.5%), respectively.