• 제목/요약/키워드: MTT method.

검색결과 420건 처리시간 0.025초

Sulfasalazine Induces Autophagic Cell Death in Oral Cancer Cells via Akt and ERK Pathways

  • Han, Hye-Yeon;Kim, Hyungwoo;Jeong, Sung-Hee;Lim, Do-Seon;Ryu, Mi Heon
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권16호
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    • pp.6939-6944
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    • 2014
  • Sulfasalazine (SSZ) is an anti-inflammatory drug that has been used to treat inflammatory bowel disease and rheumatoid arthritis for decades. Recently, some reports have suggested that SSZ also has anti-cancer properties against human tumors. However, little is known about the effects of SSZ on oral cancer. The aim of this study was to investigate the anti-cancer effects of SSZ in oral squamous cell carcinoma (OSCC) cells and to elucidate the mechanisms involved. The authors investigated the anti-proliferative effect of SSZ using the MTT method in HSC-4 cells (an OSCC cell line). Cell cycle analysis, acidic vesicular organelle (AVO) staining, monodansylcadaverine (MDC) staining and Western blotting were also conducted to investigate the cytotoxic mechanism of SSZ. SSZ significantly inhibited the proliferation of HSC-4 cells in a dose-dependent manner. In addition, SSZ induced autophagic cell death, increased microtubule-associated protein 1 light chain (MAP1-LC; also known as LC) 3-II levels, as well as induced punctate AVO and MDC staining, resulted in autophagic cell death. Furthermore, these observations were accompanied by the inhibition of the Akt pathway and the activation of ERK pathway. These results suggest that SSZ promotes autophagic cell death via Akt and ERK pathways and has chemotherapeutic potential for the treatment of oral cancer.

Antiproliferative Effects of Celecoxib in Hep-2 Cells through Telomerase Inhibition and Induction of Apoptosis

  • Zhao, Yong-Qiang;Feng, Hui-Wei;Jia, Tao;Chen, Xue-Mei;Zhang, Hui;Xu, An-Ting;Zhang, Hai-Ling;Fan, Xian-Liang
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권12호
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    • pp.4919-4923
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    • 2014
  • Background: To investigate the effect of celecoxib on telomerase activity and apoptosis in a human laryngeal squamous carcinoma cell line (Hep-2 cells). Materials and Methods: The growth inhibition rate of Hep-2 cells in vitro was measured by MTT assay, and apoptosis by TUNEL assay and flow cytometry (FCM). The TRAP-ELISA method was used to determine telomerase activity in Hep-2 cells. The mRNA expression of human telomerase RNA component(hTR), human telomerase reverse transcriptase (hTERT) and human telomerase-associated protein(hTEP1) was determined by RT-PCR assay. Expression of Bax and Bcl-2 proteins was assessed by Western blotting. Results: Celecoxib can inhibit proliferation and induce apoptosis in a dose- and time-dependent manner, repress telomerase activity, decrease hTERT mRNA and Bcl-2 protein expression and increase Bax protein expression, PGE2 had no effect on telomerase. Conclusions: Celecoxib had the antiproliferative and pro-apoptotic effect in Hep-2 cells. Apoptosis was accompanied by a decrease in telomerase activity which was directly correlated with hTERT mRNA and up-regulation of Bax/Bcl-2. Bcl-2 may thus play an important role in telomerase activity as well as apoptosis.

Combined Treatment with Stattic and Docetaxel Alters the Bax/Bcl-2 Gene Expression Ratio in Human Prostate Cancer Cells

  • Mohammadian, Jamal;Sabzichi, Mehdi;Molavi, Ommoleila;Shanehbandi, Dariush;Samadi, Nasser
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권11호
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    • pp.5031-5035
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    • 2016
  • Docetaxel, recognized as a stabilizing microtubule agent, is frequently administrated as a first line treatment for prostate cancers. Due to high side effects of monotherapy, however, combinations with novel adjuvants have emerged as an alternative strategy in cancer therapy protocols. Here, we investigated the combined effects of stattic and docetaxel on the DU145 prostate cancer cell line. Cytotoxicity was evaluated by MTT assay. To understand molecular mechanisms of stattic action, apoptotic related genes including Bcl-2, Mcl-1, Survivin and Bax were evaluated by real-time RT-PCR. Alteration in the expression of pro-apoptotic Bax and anti-apoptotic Bcl-2 genes and Bax/Bcl-2 ratio were investigated via the $2^{{\Delta}{\Delta}CT}$ method. The $IC_{50}$ values for docetaxel and stattic were $3.7{\pm}0.9nM$ and $4.6{\pm}0.8{\mu}M$, respectively. Evaluation of key gene expression levels revealed a noticeable decrease in antiapoptotic Bcl-2 and Mcl-1 along with an increase in pro-apoptotic Bax mRNA levels (p<0.05). Our results suggest that combination of a STAT3 inhibitor with doctaxel can be considered as a potent strategy for induction of apoptosis via increasing Bax mRNA expression.

Evaluation of Transferrin-Polyethylenimine Conjugate for Targeted Gene Delivery

  • Lee Kyung Man;Kim In Sook;Lee Yong Bok;Shin Sang Chul;Lee Kang Choon;Oh In Joon
    • Archives of Pharmacal Research
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    • 제28권6호
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    • pp.722-729
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    • 2005
  • With the aim to improve the specificity and to reduce the cytotoxicity of polyethylenimine (PEI), we have synthesized the conjugates of the branched PEI (25 kDa) with transferrin. The trans-ferrin-PEI (TP) conjugates with five compositions were synthesized using periodate oxidation method and confirmed by FT-IR spectroscopy and gel permeation chromatography. The free amine contents of TP conjugates, which were able to condense and deliver DNA, increased as the amount of PEI increased. TP/DNA polyplexes were characterized by measuring gel elec-trophoresis, ethidium bromide fluorescence quenching, particle size and zeta potential of complexes. Complete complexation of the polyplexes was observed above the N/P ratio of 5 in TP/DNA, and above 3 in PEI/DNA, respectively. The zeta potential of the complexes decreased as the amount of transferrin in TP conjugates increased. Transfection efficiency of TP conjugates was evaluated in HeLa cell and Jurkat cell systems. Among the five compositions of TP conjugates, TP-2 system mediated a higher $\beta$-galactosidase gene expression than PEI system in Jurkat cell which was known to express elevated numbers of transferrin receptors. From the results of the cell viability based on MTT assay, TP conjugates showed lower cytotoxicity com-pared with the PEI system. We expect that the TP conjugate can be used efficiently as a non-viral gene delivery vector.

Antioxidant, Anticancer and Anticholinesterase Activities of Flower, Fruit and Seed Extracts of Hypericum amblysepalum HOCHST

  • Keskin, Cumali
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권7호
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    • pp.2763-2769
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    • 2015
  • Background: Cancer is an unnatural type of tissue growth in which the cells exhibit unrestrained division, leading to a progressive increase in the number of dividing cells. It is now the second largest cause of death in the world. The present study concerned antioxidant, anticancer and anticholinesterase activities and protocatechuic, catechin, caffeic acid, syringic acid, p-coumaric acid and o-coumaric concentrations in methanol extracts of flowers, fruits and seeds of Hypericum amblysepalum. Materials and Methods: Antioxidant properties including free radical scavenging activity and reducing power, and amounts of total phenolic compounds were evaluated using different tests. Protocatechuic, catechin, caffeic acid, syringic acid, p-coumaric acid and o-coumaric concentrations in extracts were determined by HPLC. Cytotoxic effects were determined using the MTT test with human cervix cancer (HeLa) and rat kidney epithelium cell (NRK-52E) lines. Acetyl and butyrylcholinesterase inhibitory activities were measured by by Ellman method. Results: Total phenolic content of H. amblysepalum seeds was found to be higher than in fruit and flower extracts. DPPH free radical scavenging activity of the obtained extracts gave satisfactory results versus butylated hydroxyanisole and butylated hydroxytoluene as controls. Reducing power activity was linearly proportional to the studied concentration range: $10-500{\mu}g/mL\;LC_{50}$ values for H. amblysepalum seeds were 11.7 and 2.86 respectively for HeLa and NRK-52E cell lines. Butyryl-cholinesterase inhibitory activity was $76.9{\pm}0.41$ for seed extract and higher than with other extracts. Conclusions: The present results suggested that H. amblysepalum could be a potential candidate anti-cancer drug for the treatment of human cervical cancer, and good source of natural antioxidants.

Brine shrimp lethality and cytotoxicity assay of Araucaria bidwillii Hook in human carcinoma cell lines

  • Ahamed, KFH Nazeer;Kumar, V;Manikandan, L;Wahile, Atul M;Mukherjee, Kakali;Saha, BP;Mukherjee, Pulok K
    • Advances in Traditional Medicine
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    • 제5권1호
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    • pp.21-28
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    • 2005
  • The leaf extracts of Araucaria bidwillii Hook. (Araucariaceae) were evaluated for their cytotoxic effect in various human cancer cell lines. Preliminary investigation by brine shrimp lethality assay indicated that $LC_{50}$ value of various successive extracts were found to be less than $1000\;{\mu}g/ml$, where the ethyl acetate extract showed maximum activity of less than $100\;{\mu}g/ml$. Further cytotoxic evaluation of various leaf extracts of Araucaria bidwilli Hook was carried out in four different human cancer cell lines-acute myeloblastic leukemia (HL-60), chronic myelogenic leukemia (K-562), breast adenocarcinoma (MCF-7) and cervical epithelial carcinoma (HeLa). Cytotoxicity was assessed by trypan blue dye exclusion method and 3-(4,5-dimethyl thiazole-2yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay. From the present investigation it was found that the ethyl acetate and methanol extract of Araucaria bidwilli Hook was found to be more effective in leukemic cell lines and was less effective in MCF-7 and HeLa. The $IC_{50}$ value of the ethyl acetate extract in leukemic cell lines was found to be $28.18\;and\;34.64\;{\mu}g/ml$ and methanol extract was found to be $33.11\;&\;39.81\;{\mu}g/ml$. It can be concluded that various extract from the leaves of Araucaria bidwillii Hook. posses cytotoxic activity tested in brine shrimps and various human carcinoma cell lines.

633 nm 파장의 LED 광원이 생체 피부에 미치는 영향 (Effect of LEDs Light of 633 nm Wavelength in Skin of Organism)

  • 천민우
    • 한국전기전자재료학회논문지
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    • 제21권8호
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    • pp.760-765
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    • 2008
  • Low power laser therapy is internationally certified and is known to be effective in stimulating DNA in living organisms, increasing protein synthesis and activating cell division, smoothing blood circulation, promoting cell activation, cell regeneration and function. It also has anti-inflammatory, anti-edemic, anti-fibrous dysplastic and neuralogic hyperfunctional effects. This study was intended to verify the effect of LED irradiation therapy on wound healing in cell and animal tests by applying LED irradiator using a laser and laser diode, which was independently designed and developed to emit beams of similar wavelength to that of a laser. This equipment was fabricated using a micro-controller and a high brightness LED, and designed to enable us to control light irradiation time, intensity and reservation. In case of cell proliferation experiment, each experiment was performed to irradiation group and non-irradiation group for tissue cells. MTT assay method was chosen to verify the cell increase of two groups and the effect of irradiation on cell proliferation was examined by measuring 590 nm transmittance of micro-plate reader. In the wound healing experiment, 1$cm^2$ wounds on the skin wound of SD-Rat(Sprague-Dawley Rat) were made. Light irradiation group and none light irradiation group divided, each group was irradiated one hour a day for 9 days. As a result, the cell increase of tissue cells was verified in irradiation group as compared to non-irradiation group. And, compared with none light irradiation group, the lower incidence of inflammation and faster recovery was shown in light irradiation group.

혈소판농축혈장이 조골세포주의 세포부착에 미치는 영향에 대한 연구 (The Effect of Platelet-Rich Plasma on the Cellular Attachment of Osteoblast Cell Line)

  • 정태욱;장경수;김창회;김영수
    • 구강회복응용과학지
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    • 제19권4호
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    • pp.281-290
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    • 2003
  • Platelet-rich plasma which is made with the newly developed technique concentrating platelets 3-folds or more is also proven to be very effective method to stimulate and accelerate the healing of bone and soft tissue. This study is aimed to investigate the effect of platelet-rich plasma on the attachment of osteoblast. To evaluate the effect on human, human osteoblast cell line was cultured. Platelet-rich plasma was extracted from the blood of a healthy volunteer. The effect on the attachment was evaluated by MTT assay. To evaluate autocrine and paracrine effect on osteoblast, conditioned medium was made and compared with platelet-rich plasma. By western blot analysis, the expression of fibronectin and vitronectin in experimental groups was examined. The results were as following: The cellular attachment of osteoblast cell line increased depending on the concentration of platelet-rich plasma and conditioned medium. The amount of increasing was similar between two groups. The expression of fibronectin and vitronectin in platelet-rich plasma and conditioned medium is more than control group in western blot analysis. These findings imply that platelet-rich plasma enhance the cellular attachment by inducing fibronectin, vitronectin from osteoblast and maximize the cellular attachment by using the autocrine and paracrine effect of platelet-rich plasma.

생체용 타이타늄 합금의 산화거동 및 세포독성에 관한 연구 (A Study on Oxidation Behavior and Cytotoxicity Test of Ti-10Ta-10Nb Alloy)

  • 조홍규;이도재;이광민;이경구
    • 대한치과기공학회지
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    • 제26권1호
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    • pp.97-104
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    • 2004
  • A new Ti-10Ta-10Nb alloy has designed and examined some possibility of forming more passive oxide film by oxidation treatment which is closely related to corrosion resistance and biocompatibility. Ti-6Al-4V and Ti-10Ta-10Nb alloys were prepared by consumable vacuum arc melting and homogenized at 1050$^{\circ}C$ for 24hours. Alloy specimens were oxidized at the temperature range of 400 to 750$^{\circ}C$ for 30minutes, and the oxide films on Ti alloys were analysed by optical microscope, SEM, XPS and TGA. Cytotoxicity test was performed in MTT assay treated L929 fibroblast cell culture by indirect method. It is found out that the oxide film on Ti-10Ta-10Nb alloy is denser and thinner compared to Ti-6Al-4V alloy. The weight gain during the oxidation was increased rapidly at the temperature above 650$^{\circ}C$ for Ti-6Al-4V alloy and above 700$^{\circ}C$ for Ti-10Ta-10Nb alloy respectively. It was analysed that the passive film of the Ti alloys consisted of TiO2 through X-ray photoelectron spectroscopy (XPS) analysis. It is found out by cytotoxicity test that moderate oxidation treatment lowers cell toxicity, and Ti-10Ta-10Nb alloy showed better result compared to Ti-6Al-4V alloy.

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VVC의 기울기 기반 화면내 예측모드 결정 및 블록분할 고속화 기법 (Gradient-Based Methods of Fast Intra Mode Decision and Block Partitioning in VVC)

  • 윤용욱;박도현;김재곤
    • 방송공학회논문지
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    • 제25권3호
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    • pp.338-345
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    • 2020
  • 차세대 비디오 부호화 표준으로 진행중인 VVC(Versatile Video Coding)는 HEVC(High Efficiency Video Coding)보다 두 배 이상의 압축 성능을 달성하기 위해 다양한 기술들을 채택하고 있다. 최근 배포된 VVC 참조 SW 코덱인 VTM(VVC Test Model)은 HEVC 대비 38% 이상의 BD-rate 부호화 성능 향상을 보이는 반면 부호화와 복호화 복잡도가 각각 9배, 2배 정도 증가를 보인다. 특히, 재귀적 MTT(Multi-Type Tree) 분할 구조와 HEVC 대비 2배로 증가한 화면내 예측모드 수로 인해 상당한 부호화기의 복잡도가 증가하였으며, 이를 감소시키기 위한 다양한 기법들이 연구되고 있다. 본 논문에서는 부호화기의 복잡도를 감소시키기 위하여 블록내 화소의 기울기를 이용한 고속 화면내 예측모드 결정 및 블록분할 기법을 제시한다. 실험결과 VTM6.0 대비 AI(All Intra) 부호화 구조에서 3.54%의 부호화 성능 감소와 65%의 부호화 시간 절감 효과를 얻었다.