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Plumbagin from Plumbago Zeylanica L Induces Apoptosis in Human Non-small Cell Lung Cancer Cell Lines through NF-κB Inactivation

  • Xu, Tong-Peng;Shen, Hua;Liu, Ling-Xiang;Shu, Yong-Qian
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.4
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    • pp.2325-2331
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    • 2013
  • Objective: To detect effects of plumbagin on proliferation and apoptosis in non-small cell lung cancer cell lines, and investigate the underlying mechanisms. Materials and Methods: Human non-small cell lung cancer cell lines A549, H292 and H460 were treated with various concentrations of plumbagin. Cell proliferation rates was determined using both cell counting kit-8 (CCK-8) and clonogenic assays. Apoptosis was detected by annexin V/propidium iodide double-labeled flow cytometry and TUNEL assay. The levels of reactive oxygen species (ROS) were detected by flow cytometry. Activity of NF-${\kappa}B$ was examined by electrophoretic mobility shift assay (EMSA) and luciferase reporter assay. Western blotting was used to assess the expression of both NF-${\kappa}B$ regulated apoptotic-related gene and activation of p65 and $I{\kappa}B{\kappa}$. Results: Plumbagin dose-dependently inhibited proliferation of the lung cancer cells. The IC50 values of plumbagin in A549, H292, and H460 cells were 10.3 ${\mu}mol/L$, 7.3 ${\mu}mol/L$, and 6.1 ${\mu}mol/L$ for 12 hours, respectively. The compound concentration-dependently induced apoptosis of the three cell lines. Treatment with plumbagin increased the intracellular level of ROS, and inhibited the activation of NK-${\kappa}B$. In addition to inhibition of NF-${\kappa}B$/p65 nuclear translocation, the compound also suppressed the degradation of $I{\kappa}B{\kappa}$. ROS scavenger NAC highly reversed the effect of plumbagin on apoptosis and inactivation of NK-${\kappa}B$ in H460 cell line. Treatment with plumbagin also increased the activity of caspase-9 and caspase-3, downregulated the expression of Bcl-2, upregulated the expression of Bax, Bak, and CytC. Conclusions: Plumbagin inhibits cell growth and induces apoptosis in human lung cancer cells through an NF-${\kappa}B$-regulated mitochondrial-mediated pathway, involving activation of ROS.

The Effect of Irradiation and Epidermal Growth Factor on Cell Cycle and Apoptosis Induction in Human Epithelial Tumor Cell Lines (수 종의 상피기원 종양 세포주에서 방사선 조사와 표피성장인자 투여에 따른 세포 주기의 변화와 apoptosis 유발에 관한 연구)

  • Han Won-Jeong;Heo Min-Suk;Lee Sam-Sun;Choi Soon-Chul;Park Tae-Won
    • Imaging Science in Dentistry
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    • v.30 no.1
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    • pp.71-79
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    • 2000
  • Purpose : This study was aimed to evaluate the cell cycle arrest and apoptosis induction after irradiation and epidermal growth factor (EGF) treatment in three human epithelial tumor cell lines (A431, Siha, KB). Materials and Methods: Single irradiation of 2, 5 and 10 Gy was done on three cell lines with 5.38 Gy/min dose rate using Cs-137 irradiator at room temperature. Also, EGF of 10 ng/ml was added immediately after 10 Gy irradiation. Cell growth was evaluated by counting the living cell number using a hemocytometer at 1 day, 2 days, 3 days, 4 days and 5 days after irradiation. Cell cycle arrest and apoptosis induction were assayed with the flow cytometry at 8 hours, 12 hours, 1 day, 2 days, 3 days, 4 days and 5 days after irradiation. Results : Growth of irradiated three cell lines were inhibited in proportion to radiation dose. EGF treatment after irradiation showed various results according to cell lines. On all cell lines, G2 arrest was detected after 8 hours and maximized after 12 hours or 1 day. Amount of G2 arrest was positively dose dependent. However, EGF showed no significant change on G2 arrest. G2 arrest was recovered with time at 2 Gy and 5 Gy irradiation. However, at 10 Gy irradiation, G2 arrest was continued. Apoptosis was detected at 10 Gy irradiation. On EGF treated group after irradiation, A431 and Siha cell lines showed slightly increased apoptosis but there was no statistically significant difference. KB cell line showed no marked change of apoptosis induction. Conclusion : Irradiation effects on cell cycle arrest and apoptosis induction in three human epithelial tumor cell lines, however epidermal growth factor doesn't effect on.

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Down-regulation of Phosphoglucose Isomerase/Autocrine Motility Factor Enhances Gensenoside Rh2 Pharmacological Action on Leukemia KG1α Cells

  • You, Zhi-Mei;Zhao, Liang;Xia, Jing;Wei, Qiang;Liu, Yu-Min;Liu, Xiao-Yan;Chen, Di-Long;Li, Jing
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1099-1104
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    • 2014
  • Aims and Background: Ginsenoside Rh2, which exerts the potent anticancer action both in vitro and in vivo, is one of the most well characterized ginsenosides extracted from ginseng. Although its effects on cancer are significant, the underlying mechanisms remain unknown. In this study, we sought to elucidate possible links between ginsenoside Rh2 and phosphoglucose isomerase/autocrine motility factor (PGI/AMF). Methods: $KG1{\alpha}$, a leukemia cell line highly expressing PGI/AMF was assessed by western blot analysis and reverse transcription- PCR (RT-PCR) assay after transfection of a small interfering (si)-RNA to silence PGI/AMF. The effect of PGI/AMF on proliferation was measured by typan blue assay and antibody array. A cell counting kit (CCK)-8 and flow cytometry (FCM) were adopted to investigate the effects of Rh2 on PGI/AMF. The relationships between PGI/AMF and Rh2 associated with Akt, mTOR, Raptor, Rag were detected by western blot analysis. Results: KG1${\alpha}$ cells expressed PGI/AMF and its down-regulation significantly inhibited proliferation. The antibody array indicated that the probable mechanism was reduced expression of PARP, State1, SAPK/JNK and Erk1/2, while those of PRAS40 and p38 were up-regulated. Silencing of PGI/AMF enhanced the sensibility of $KG1{\alpha}$ to Rh2 by suppressing the expression of mTOR, Raptor and Akt. Conclusion: These results suggested that ginsenoside Rh2 suppressed the proliferation of $KG1{\alpha}$, the same as down-regulation of PGI/AMF. Down-regulation of PGI/AMF enhanced the pharmacological effects of ginsenoside Rh2 on KG1${\alpha}$ by reducing Akt/mTOR signaling.

KCl Mediates $K^+$ Channel-Activated Mitogen-Activated Protein Kinases Signaling in Wound Healing

  • Shim, Jung Hee;Lim, Jong Woo;Kim, Byeong Kyu;Park, Soo Jin;Kim, Suk Wha;Choi, Tae Hyun
    • Archives of Plastic Surgery
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    • v.42 no.1
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    • pp.11-19
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    • 2015
  • Background Wound healing is an interaction of a complex signaling cascade of cellular events, including inflammation, proliferation, and maturation. $K^+$ channels modulate the mitogen-activated protein kinase (MAPK) signaling pathway. Here, we investigated whether $K^+$ channel-activated MAPK signaling directs collagen synthesis and angiogenesis in wound healing. Methods The human skin fibroblast HS27 cell line was used to examine cell viability and collagen synthesis after potassium chloride (KCl) treatment by Cell Counting Kit-8 (CCK-8) and western blotting. To investigate whether $K^+$ ion channels function upstream of MAPK signaling, thus affecting collagen synthesis and angiogenesis, we examined alteration of MAPK expression after treatment with KCl (channel inhibitor), NS1619 (channel activator), or kinase inhibitors. To research the effect of KCl on angiogenesis, angiogenesis-related proteins such as thrombospondin 1 (TSP1), anti-angiogenic factor, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), pro-angiogenic factor were assayed by western blot. Results The viability of HS27 cells was not affected by 25 mM KCl. Collagen synthesis increased dependent on time and concentration of KCl exposure. The phosphorylations of MAPK proteins such as extracellular-signal-regulated kinase (ERK) and p38 increased about 2.5-3 fold in the KCl treatment cells and were inhibited by treatment of NS1619. TSP1 expression increased by 100%, bFGF expression decreased by 40%, and there is no significant differences in the VEGF level by KCl treatment, TSP1 was inhibited by NS1619 or kinase inhibitors. Conclusions Our results suggest that KCl may function as a therapeutic agent for wound healing in the skin through MAPK signaling mediated by the $K^+$ ion channel.

Bacterial Contamination of Dental Unit Water Systems in a Student Clinical Simulation Laboratory of College of Dentistry (치과대학 임상 시뮬레이션 실습실 치과용 유니트 수계의 세균 오염도 조사)

  • Yoon, Hye Young;Lee, Si Young
    • Journal of dental hygiene science
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    • v.15 no.2
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    • pp.232-237
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    • 2015
  • The water supplied from dental unit water systems (DUWS) in dentistry may be heavily contaminated with bacteria and thus may be a potential source of infection for both practice staff and patients. The aim of this study was to evaluate the level of heterotrophic bacteria and to confirm the presence of opportunistic pathogens from DUWS in student clinical simulation laboratory of college of dentistry. Water samples were collected from 36 ultrasonic scalers in student clinical simulation laboratory. The levels of heterotrophic bacteria in water samples were quantified by counting colony forming units (CFUs) on R2A agar media. In addition, opportunistic pathogens were detected by using the polymerase chain reaction (PCR) method. The mean CFUs were 16,095 CFU/ml for water samples and all of water samples exceeded current American Dental Association recommendations of 200 CFU/ml. Pseudomonas species and non-tuberculous Mycobacterium species were detected in the one sample and two samples, respectively, among the 36 water samples by the PCR with specific primers for these bacteria. Our study indicated that DUWS in student clinical simulation laboratory can cause potential infection in students and participants. This study suggested the dental unit water line management and wearing personal protective equipment in student clinical simulation laboratory will be needed to reduce bacterial contamination.

A Study on Effect of Temperature on Particle Size Distribution of Nickel Ferrite (온도의 영향에 따른 니켈페라이트의 입자 크기 분포 연구)

  • Ahn, Hyung-Kyoung;Lee, In-Hyoung;Jeong, Hyun-Jun;Park, Byung-Gi
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.9 no.6
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    • pp.1768-1774
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    • 2008
  • The particulate behaviors of nickel ferrite were investigated under the simulated PWR shutdown chemistry conditions. Temperature of the simulated water with concentration of 0.1 ppm Li and 2,000 ppm B was dropped from $300^{\circ}C$ to $150^{\circ}C$ with a rate of $0.625^{\circ}C/min$ and then constantly maintained at $150^{\circ}C$ under the pressure of 2,500 psi. The on-line particle counting and the concentration measurement of nickel dissolved were performed under 5, 15 and 25 cc/kg $H_2O$ dissolved hydrogen. Experimental results showed that total particle count in the simulated water was not greatly changed for three hydrogen concentrations as temperature was decreased. However, particles were smaller as temperature was decreased and then maintained constantly. The degree of variation in particle size distribution was greater at 15 cc/kg $H_2O$ dissolved hydrogen than any other dissolved hydrogen concentrations. Concentration of nickel ion was increased as temperature was decreased and was higher at 15 cc/kg $H_2O$ dissolved hydrogen than any other dissolved hydrogen concentrations. Theses results show that nickel ferrite is unstable with temperature variation and at dissolved hydrogen concentration of 15 cc/kg $H_2O$.

Effect of serotonin on the cell viability of the bovine mammary alveolar cell-T (MAC-T) cell line

  • Xusheng, Dong;Chen, Liu;Jialin, Miao;Xueyan, Lin;Yun, Wang;Zhonghua, Wang;Qiuling, Hou
    • Journal of Animal Science and Technology
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    • v.64 no.5
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    • pp.922-936
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    • 2022
  • 5-Hydroxytryptamine (5-HT), a monoamine, as a local regulator in the mammary gland is a chemical signal produced by the mammary epithelium cell. In cows, studies have shown that 5-HT is associated with epithelial cell apoptosis during the degenerative phase of the mammary gland. However, studies in other tissues have shown that 5-HT can effectively promote cell viability. Whether 5-HT could have an effect on mammary cell viability in dairy cows is still unknown. The purpose of this study was to determine: (1) effect of 5-HT on the viability of bovine mammary epithelial cells and its related signaling pathways, (2) interaction between prolactin (PRL) and 5-HT on the cell viability. The bovine mammary alveolar cell-T (MAC-T) were cultured with different concentrations of 5-HT for 12, 24, 48 or 72 hours, and then were assayed using cell counting kit-8, polymerase chain reaction (PCR) and immunobloting. The results suggested that 20 μM 5-HT treatment for 12 or 24 h promote cell viability, which was mainly induced by the activation of 5-HT receptor (5-HTR) 1B and 4, because the increase caused by 5-HT vanished when 5-HTR 1B and 4 was blocked by SB224289 and SB204070. And protein expression of mammalian target of rapamycin (mTOR), eukaryotic translation elongation factor 2 (eEF2), janus kinase 2 (JAK2) and signal transducer and activator of transcription 5 (STAT5) were decreased after blocking 5-HT 1B and 4 receptors. When MAC-T cells were treated with 5-HT and PRL simultaneously for 24 h, both the cell viability and the level of mTOR protein were significantly higher than that cultured with 5-HT or PRL alone. In conclusion, our study suggested that 5-HT promotes the viability of MAC-T cells by 5-HTR 1B and/or 4. Furthermore, there is a reciprocal relationship between PRL and 5-HT.

DoS/DDoS attacks Detection Algorithm and System using Packet Counting (패킷 카운팅을 이용한 DoS/DDoS 공격 탐지 알고리즘 및 이를 이용한 시스템)

  • Kim, Tae-Won;Jung, Jae-Il;Lee, Joo-Young
    • Journal of the Korea Society for Simulation
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    • v.19 no.4
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    • pp.151-159
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    • 2010
  • Currently, by using the Internet, We can do varius things such as Web surfing, email, on-line shopping, stock trading on your home or office. However, as being out of the concept of security from the beginning, it is the big social issues that malicious user intrudes into the system through the network, on purpose to steal personal information or to paralyze system. In addition, network intrusion by ordinary people using network attack tools is bringing about big worries, so that the need for effective and powerful intrusion detection system becomes very important issue in our Internet environment. However, it is very difficult to prevent this attack perfectly. In this paper we proposed the algorithm for the detection of DoS attacks, and developed attack detection tools. Through learning in a normal state on Step 1, we calculate thresholds, the number of packets that are coming to each port, the median and the average utilization of each port on Step 2. And we propose values to determine how to attack detection on Step 3. By programing proposed attack detection algorithm and by testing the results, we can see that the difference between the median of packet mounts for unit interval and the average utilization of each port number is effective in detecting attacks. Also, without the need to look into the network data, we can easily be implemented by only using the number of packets to detect attacks.

Studies on the Radiation Induced Apoptosis by Morphological and Biochemical Analysis in A431 Cells (방사선에 조사된 인체세포주(A431)에서 형태학적 분석법과 생화학적 분석법을 이용한 방사선 유도 세포고사에 관한 연구)

  • Choi, Keun-Hee;Bom, Hee-Seung;Kim, Ji-Yeul
    • The Korean Journal of Nuclear Medicine
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    • v.33 no.3
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    • pp.306-315
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    • 1999
  • Purpose: We performed this study to evaluate the process of radiation induced apoptosis in A431 skin epithelial cancer cell line. Materials and Methods: Low to high dose radiation (0, 2, 5, 10, 25 Gy) was given to A431 cells by Cs-137 cell irradiator. Apoptosis was evaluated by cell morphology, dye exclusion test, and DNA laddering. Results: Cell viability decreased as the radiation dose increased. Number of apoptotic bodies increased as radiation dose increased. It increased most significantly at 12 hours after irradiation. Lactate dehydrogenase activity in culture medium increased according to radiation dose and time after irradiation. DNA ladders could be identified in irradiated cells, but, it had no correlation with radiation dose or time after irradiation. Conclusion: Radiation-induced apoptosis which was the main course of cell death in A431 cells could be analyzed quantitatively by counting apoptotic bodies under microscope. Apoptosis increased as radiation dose increased.

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The effects of physical factors in SPECT (물리적 요소가 SPECT 영상에 미치는 영향)

  • 손혜경;김희중;나상균;이희경
    • Progress in Medical Physics
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    • v.7 no.1
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    • pp.65-77
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    • 1996
  • Using the 2-D and 3-D Hoffman brain phantom, 3-D Jaszczak phantom and Single Photon Emission Computed Tomography, the effects of data acquisition parameter, attenuation, noise, scatter and reconstruction algorithm on image quantitation as well as image quality were studied. For the data acquisition parameters, the images were acquired by changing the increment angle of rotation and the radius. The less increment angle of rotation resulted in superior image quality. Smaller radius from the center of rotation gave better image quality, since the resolution degraded as increasing the distance from detector to object increased. Using the flood data in Jaszczak phantom, the optimal attenuation coefficients were derived as 0.12cm$\^$-1/ for all collimators. Consequently, the all images were corrected for attenuation using the derived attenuation coefficients. It showed concave line profile without attenuation correction and flat line profile with attenuation correction in flood data obtained with jaszczak phantom. And the attenuation correction improved both image qulity and image quantitation. To study the effects of noise, the images were acquired for 1min, 2min, 5min, 10min, and 20min. The 20min image showed much better noise characteristics than 1min image indicating that increasing the counting time reduces the noise characteristics which follow the Poisson distribution. The images were also acquired using dual-energy windows, one for main photopeak and another one for scatter peak. The images were then compared with and without scatter correction. Scatter correction improved image quality so that the cold sphere and bar pattern in Jaszczak phantom were clearly visualized. Scatter correction was also applied to 3-D Hoffman brain phantom and resulted in better image quality. In conclusion, the SPECT images were significantly affected by the factors of data acquisition parameter, attenuation, noise, scatter, and reconstruction algorithm and these factors must be optimized or corrected to obtain the useful SPECT data in clinical applications.

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