• 제목/요약/키워드: MTS Assay

검색결과 183건 처리시간 0.029초

Prediction of Chemotherapeutic Response in Unresectable Non-small-cell Lung Cancer (NSCLC) Patients by 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) Assay

  • Chen, Juan;Cheng, Guo-Hua;Chen, Li-Pai;Pang, Ting-Yuan;Wang, Xiao-Le
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권5호
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    • pp.3057-3062
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    • 2013
  • Background: Selecting chemotherapy regimens guided by chemosensitivity tests can provide individualized therapies for cancer patients. The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium, inner salt (MTS) assay is one in vitro assay which has become widely used to evaluate the sensitivity to anticancer agents. The aim of this study was to evaluate the clinical applicability and accuracy of MTS assay for predicting chemotherapeutic response in unresectable NSCLC patients. Methods: Cancer cells were isolated from malignant pleural effusions of patients by density gradient centrifugation, and their sensitivity to eight chemotherapeutic agents was examined by MTS assay and compared with clinical response. Results: A total of 37 patients participated in this study, and MTS assay produced results successfully in 34 patients (91.9%). The sensitivity rates ranged from 8.8% to 88.2%. Twenty-four of 34 patients who received chemotherapy were evaluated for in vitro-in vivo response analysis. The correlation between in vitro chemosensitivity result and in vivo response was highly significant (P=0.003), and the total predictive accuracy, sensitivity, specificity, positive predictive value, and negative predictive value for MTS assay were 87.5%, 94.1%, 71.4%, 88.9%, and 83.3%, respectively. The in vitro sensitivity for CDDP also showed a significant correlation with in vivo response (P=0.018, r=0.522). Conclusion: MTS assay is a preferable in vitro chemosensitivity assay that could be use to predict the response to chemotherapy and select the appropriate chemotherapy regimens for unresectable NSCLC patients, which could greatly improve therapeutic efficacy and reduce unnecessary adverse effects.

꽃송이버섯 추출물이 RAW 264.7 세포에서 TNF-α, iNOS, IL-1β 유전자 발현에 미치는 영향 (Effect of Water Extract of Sparasis crispa on the Expression of TNF-α, iNOS and IL-1β Genes in RAW 264.7 Cells)

  • 한효상
    • 대한통합의학회지
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    • 제9권1호
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    • pp.163-171
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    • 2021
  • Purpose : The purpose of this study was to examine the anti-inflammatory effects of Sparassis crispa (SC). SC is a well-known traditional herbal remedy and its mushroom is used for treatment of inflammation. Many diseases that are increasing recently have characteristics of inflammatory diseases. Researchers are finding bioactive substances from natural products that can promote treatment and prevention of inflammation. We investigated the effect of water extracted from SC on the expression of effector genes involved in the function of RAW 264.7 cells. Methods : Effects of RAW 264.7 cells on cell viability, antioxidation, and mRNA expression were examined using water extracts from SC. A 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed to determine the effect of water extracts from SC on cell viability in RAW 264.7 cells. Inflammation of RAW 264.7 cells induced by lipopolysaccharide (LPS) treatment and expression levels of inflammatory cytokine TNF-α, iNOS and IL-1β gene were analyzed using quantitative reverse transcription PCR (qRT-PCR) analysis. Results : The MTS assay was performed on RAW 264.7 cells after treatment with various concentrations of water extracts of SC. Treatment of RAW 264.7 cells with water extracts from SC and LPS at a concentration of 0.125, 0.5 mg/㎖ for twenty four hours promoted mRNA expression of TNF-α, iNOS and IL-1β. Conclusion : MTS assay was applied to RAW 264.7 cells after various concentrations of water extracts of SC. Through experimental demonstration of anti-oxidant and anti-inflammatory effects of water extracts from SC, we suggest that SC is a valuable material for the prevention and treatment of various inflammatory diseases.

LED조사가 인간 피부 각질세포의 증식에 미치는 융복합적인 영향 (Effect of LED Irradiation on Proliferation of Human Epidermal Keratinocyte for Convergence)

  • 박정숙;김미혜;이재혁
    • 디지털융복합연구
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    • 제14권11호
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    • pp.639-644
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    • 2016
  • 본 연구의 목적은 발광 다이오드 (LED)을 인간 피부 각질 세포에 조사 시 콜라겐, 프로 콜라겐의 증식 발현을 조사하기 위해 실시되었었다. LED 조사 시 안전하게 인간의 피부에 적용할 수 있는지 여부를 결정하기 위해, LED 조사의 증식 효과는 인간 표피 각질세포에서 MTS 분석으로 결정하였다. 470nm의 파장 조사는 세포 독성 없이 mRNA의 콜라겐의 발현, 프로 콜라겐을 증가시켰으며, 이 결과는 470nm LED 조사가 피부각질세포 증식 효과와 콜라겐 합성에 영향을 미칠 수 있음을 시사한다. 또한 LED 조사시 독성 효과는 인간 피부 섬유 아세포 (HDF)에서 MTS 분석으로 결정 한 결과 세포 증식에 독성을 나타내지 않았다. 470nm LED 조사시 시험 관내 콜라겐 합성 활동을 증가시킴으로 피부미용 및 융복합적인 부분에 활용할 수 있는 기초 자료로 활용이 가능하다고 사료된다.

상황버섯균사체 쌀의 HT22 신경세포 보호 효과 (Neuroprotective Effect of Rice with Phellinus linteus Mycelium in HT22 Cells)

  • 김지현;전순실
    • 한국식품영양과학회지
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    • 제46권7호
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    • pp.886-890
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    • 2017
  • 상황버섯균사체 쌀의 뇌세포 보호 효과를 분석하여 상황버섯균사체 쌀에 대한 기초자료를 제공하고, 이를 이용하여 뇌 질환의 예방과 관련된 기능성 식품으로의 개발가능성을 찾고자 하였다. 상황버섯균사체 쌀 추출물을 처리하여 마우스 해마세포주 HT22의 세포보호 효과를 알아보기 위해 MTS assay를 이용한 세포생존율과 western blot을 이용한 세포사멸 단백질의 발현을 관찰하였다. 상황버섯균사체 쌀 추출물의 단일 독성을 측정한 결과 상황버섯균사체 쌀 추출물 처리군은 102.68% 이상의 생존율을 보여 신경세포의 사멸에 영향을 주지 않았다. 또한, 상황버섯균사체 쌀 추출물과 $H_2O_2$를 함께 처리하였을 때 $H_2O_2$로 자극된 세포는 63.80%의 세포생존율을 나타내었고, 상황버섯균사체 쌀 추출물과 $H_2O_2$가 함께 처리된 세포는 76.85%, 92.46%, 95.57%로 농도 의존적으로 세포생존율이 증가하였다. HT22 cell에 $H_2O_2$와 상황버섯균사체 쌀 추출물을 처리하여 apoptosis 유도 단백질을 측정한 결과, pro-apoptotic protein인 Bax는 발현이 억제되었고, anti-apoptotic protein인 Bcl-2는 발현이 증가함을 확인하였다. 또한, caspase-3와 PARP의 발현을 억제하여 상황버섯균사체 쌀 추출물이 $H_2O_2$로 유도되는 apoptosis를 억제하는 것을 확인할 수 있었다.

A Study on the Mechanism of Oxidative Stress, Screening of Protective Agents and Signal Transduction of Cell Differentiation in Cultured Osteoblast and Osteoclast Damaged by Reactive Oxygen Species

  • Park Seung-Taeck;Jeon Seung-Ho
    • 대한의생명과학회지
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    • 제11권3호
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    • pp.319-326
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    • 2005
  • It is well known that oxidative stress of reactive oxygen species (ROS) may be a causative factor in the pathenogenesis of bone disorder on osteoblast or osteoclast. The purpose of this study was to evaluate the cytotoxicity of oxidative stress, protective effect of glutamate receptor antagoinst against ROS-induced osteotoxicity, secretion of tumor necrosis factor $(TNF)-\alpha$ and the expression of c-fos gene in the cultured rat osteoblasts and osteoclasts. Cell viability by MTS assay or !NT assay, activity of glutathione peroxidase (GPx), lipid peroxidation (LPO) activity, protein synthesis by sulforhodamine B (SRB) assay, alkaline phosphatase (ALP) activity, lactate dehydrogenase (LDH) activity, MTS assay for NMDA (N-methyl-D-aspartate) receptor antagonist or AMPA/kainate receptor antagonist, measurement for $TNF-\alpha$, and c-fos gene expression were performed after these cells were treated with or without various cocentrations of xanthine oxidase (XO), hypoxanthine (HX), D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX), respectively. In this study, XO/HX showed decreased cell viability and glutathione peroxidase (GPx) activity, but it showed increased LPO activity, $TNF-\alpha$ secretion and c-fos expression. APV and CKA incresed protein sythesis and ALP activity. While, CNQX or DNQX did not show any protective effect in LDH activity or cell viability. From these results, XO/HX showed cytotoxic effect in cultured rat osteoblast or osteoclast, and also NMDA receptor antagonist such as APV or CKA was effective in blocking XO/HX-induced osteotoxicity in these cultures.

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강활유풍탕(羌活愈風湯)이 뇌허혈(腦虛血)로 유도된 대뇌신경세포손상(大腦神經細胞損傷)에 미치는 영향 (Effects of Kangwhalyupung-tang on the Cerebral Neuronal Damage induced by Ischemia)

  • 문병순;황충연;김경요;이건목;성강경;이대용;이성근
    • 대한한방내과학회지
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    • 제24권1호
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    • pp.11-20
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    • 2003
  • Objective : Experimental studies have been done to elucidate the effect of kangwhalyupung-tang(KWYPT) on neuronal cell damage induced by brain ischemia. Method : The cytotoxic effect of ischemia was measured in the MTS assay cultures. MTS assay, INT assay, neurofilament(NF) enzymeimmuno assay(EIA). And the KWYPT on ischemia-induced neurotoxicity were examined by in vitro assays. Results : 1. The KWYPT protected effectively neuronal cell-death resulted from brain ischemia induced by the treatment of $95%N_2/5%CO_2$ for 10 min in those dependent fashion. 2. The KWYPT effectively increased the amount of NF resulting from brain ischemia, induced by the treatment of $95%N_2/5%CO_2$ for 15 min in those dependent fashions. Conclusions : KWYPT protects the brain ischemia-induced neurotoxicity through the increase of cell viability and of neurofilament in dose-dependent manner.

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자초(紫草) 열수추출물이 각질형성세포 HaCaT의 세포 연접 관련 유전자의 발현에 미치는 영향 연구 (Investigation of the effect of Lithospermi Radix on tight-junction related genes in HaCaT cells)

  • 조남준;이병권;이웅희;김기광;김균언;한효상
    • 대한본초학회지
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    • 제32권3호
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    • pp.55-61
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    • 2017
  • Objectives : The aim of this research was to determine the diverse effects of Lithospermi Radix Water Extract (LR) on human keratinocyte HaCaT cells, and to examine whether those effects could be applied to the human skin. Methods : We examined effect of LR on the cell viability of using the MTS assay in human keratinocyte HaCaT cells. The antioxidation effect of LR was analyzed relative to the well-known antioxidant resveratrol, using an ABTS assay. Quantitative RT-PCR analysis revealed that, in HaCaT cells, LR influenced the mRNA expression of tight-junction genes associated with skin moisturization. Furthermore, a wound-healing assay demonstrated altered cell migration in LR-treated HaCaT cells. Result : The cytotoxicity was confirmed to be higher in LR at a concentration of $800{\mu}g/m{\ell}$ using the MTS assay in HaCaT cells. In comparison to $100{\mu}M$ resveratrol, $1,600{\mu}g/m{\ell}$ LR showed either a similar or superior antioxidation effect. LR treatment in HaCaT cells reduced the mRNA expression levels of claudin 3, claudin 4, claudin 6, claudin 8, and ZO-2 to less than 0.80-fold, whereas JAM-A and Tricellulin mRNA expression level increased more than 1.33-fold. In addition, HaCaT cells migration was decreased to 83.9% by LR treatment. Conclusions : LR of antioxidation activity will have an anti-aging effect on the skin by reducing oxidative stress. Further studies are required to address the implications for human skin, given LR's effects of altering mRNA expression of tight junction-related gene and decreasing cell migration of HaCaT cells.

블루베리 열수 추출물의 근아세포의 근분화에 미치는 영향 (Investigation of the effect of Blueberry hydrothermal extracts on myoblast differentiation)

  • 최영수;김은미;최선경;이웅희;한효상;김기광
    • 대한본초학회지
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    • 제35권3호
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    • pp.25-32
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    • 2020
  • Objectives : At present, aging-related degenerative muscle diseases are considered a serious problem. However, the effects on muscles regarding the efficacy of blueberry have not been studied. In this study, we tried to find out the correlation between blueberry and muscle. Methods : 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay was performed to confirm the antioxidant efficacy of blueberry hydrothermal extract. To determine the effect of blueberry hydrothermal extracts (BHE) on myoblast activity, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed. To confirm the effect of blueberry hydrothermal extracts on the differentiation of myoblast into myotubes, protein expression levels of myosin heavy chain 3 (Myh3) and paired box 3/7 (pax3/7) were confirmed by immunoblot analysis. In addition, immunofluorescence microscopy was performed to confirm the effect on myotube formation of blueberry hydrothermal extracts. Results : Antioxidative efficacy and low toxicity were confirmed through ABTS assay and MTS assay of blueberry extract for myoblasts. As a result of immunoblot analysis and immunofluorescence analysis, the decrease in myogenic marker Pax3/7 was not confirmed, but myotubes The specific expression inhibitory activity of the forming protein Myh3 was confirmed. Through this, it was confirmed that the blueberry extract has a negative activity against myoblast differentiation. Conclusion : This experiment confirmed that blueberry hydrothermal extract has excellent antioxidant efficacy and negative results in inhibiting the differentiation and proliferation of myoblast. This requires deep study of certain ingredients and requires reassessment of the dietary intake of blueberries.

홍화자와 두충 혼합 추출물이 MG-63 조골세포의 Alkaline Phosphatase 활성에 미치는 영향 (Effects of Alkaline Phosphatase Activity on the Extract of Carthami Semen and Eucommiae Cortex in Human Osteoblast-like MG-63 Cell Line)

  • 심재근;이재혁;여명구;박정숙
    • 동아시아식생활학회지
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    • 제23권1호
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    • pp.39-43
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    • 2013
  • Carthamus tinctorius L. and Eucommia umoides Oliver are often used in traditional herbal medicines for reducing damage to the liver, kidney, bone and muscle. In the present study, we investigated cell viability and alkaline phosphatase activity in the human osteoblast-like MG-63 cell line with methanol extracts of Carthami Semen (CS) and Eucommiae Cortex (EC) alone or in a mixture (CS+EC). Osteoblast cell viability was evaluated using the MTS assay and alkaline phosphatase activity assays. The cell viability and alkaline phosphatase activity significantly increased in MG-63 osteoblast cells treated with the CS+EC mixture. These findings suggest the CS+EC mixture may have beneficial effects on bone health through the proliferation of osteoblast cells.

갈근(葛根)의 착즙 및 열수 추출에 따른 모유두 세포의 모발 성장 관련 mRNA 발현에 미치는 영향 (Investigation of the differential effect of juice or water extract from Puerariae Radix on hair growth related-genes in dermal papilla cells)

  • 전해리;조남준;김기광;한효상
    • 대한본초학회지
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    • 제33권1호
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    • pp.1-7
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    • 2018
  • Objectives : Puerariae Radix extracts are a rich source of isoflavones that affect the activity of hepatic antioxidant enzymes. However, the effect of these extracts on hair growth in human dermal papilla (DP) cells is unknown. The purpose of this study is to compare the effects between juice and water extracts of Puerariae Radix on the mRNA expression levels of hair growth-related genes in dermal papilla (DP) cells. Methods : The antioxidant activity of juice and water extracts of Puerariae Radix was analyzed using an ABTS assay. The cytotoxicity was analyzed using the MTS assay in DP cells. mRNA expression levels of hair growth-related genes in dermal papilla (DP) cells were analyzed using quantitative RT-PCR analysis. Results : Juice and water extracts of Puerariae Radix showed strong antioxidant activity. The cytotoxicity was confirmed to be higher in the juice extract than the water extract, using the MTS assay on DP cells. The mRNA expression levels of CTNNB1, FGF7, and BMP6 were significantly increased after treatment with water extract, whereas the juice extract did not affect the expression of hair growth-related genes. Conclusions : Our study provides evidence that water extract of Puerariae Radix is effective at inducing hair growth, by promoting the mRNA expression levels of hair growth-related genes. Also in the future, studies should be conducted to investigate the effects of Puerariae radix extracts on the various hair growth mechanisms of dermal papilla cells.