• Title/Summary/Keyword: Chk2

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Effect of Benzo[a]pyrene on Genes Related to the Cell Cycle and Cytochrome P450 of Saccharomyces cerevisiae

  • Lee, Hyun-Joo;Gu, Man-Bock
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.624-627
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    • 2003
  • Benzo[a]pyrene (B[a]P) is an environmental pollutant that has been implicated in carcinogenesis. Saccharomyces cerevisiae was treated with B[a]P, and the responses of its cytochrome P450 (CYP) enzyme and DNA-damage checkpoint genes were examined through gene expression profiles using a reverse transcription polymerase chain reaction (RT-PCR). The DNA-damage checkpoint genes tested were the chk1 and pds1 genes, involved in a metaphase arrest, the swi6 gene targeted by G1 arrest, the pol2 gene related to S phase arrest, and the cln2 gene encoding a cyclin protein, all of which are based on rad9 and rad24. Among these genes, no noticeable effect was found when the cells were exposed to various concentrations of B[a]P. However, the transcriptional activity of CYP51 was significantly different when the cells were exposed to B[a]P. Accordingly, the present results indicate that cytochrome P450 plays a more significant role than DNA-damage checkpoint genes in the response of S. cerevisiae to B[a]P.

Effect of the Ethanol Extract of Vitis labrusca Root on Apoptosis in Hep G2 Cells (포도근 에탄올 추출물이 Hep G2 세포의 자연사에 미치는 효과)

  • Lee, Dong-Kyo;Lee, Kang-Pa;Kim, Hyuck;Choi, Byung-Jin;Chang, Hae-Ryong;Park, Won-Hwan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.2
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    • pp.377-384
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    • 2008
  • The root of Vitis labrusca, is used as a source of health promoting drug in Korean traditional medicine. It has been reported that root of Vitis labrusca has antioxidant, anti lipid peroxidation and anti-reactive nitrogen species (RNS) activities. The aim of this study was to elucidate the molecular changes of apoptotic signaling pathways in phorbol 12-myristate 13 acetate (PMA)-induced human hepatocellular carcinoma cell line (Hep G2). The root of Vitis labrusca, ethanol extract (RVLEE) was tested for cell viability on Hep G2 cell using the MTT assay. RVLEE exhibited weak cytotoxic activity. However, treatment of Hep G2 cells with RVLEE suppressed PMA-induced cell proliferation. Also, dramatic changes of cell death signals in cellular molecules such as Chk2/Cds1, CIDE-B, CLIMP-63, Bax, Bcl-xL, C-myc, Bcl-2, Bric-5, NIP-3, TRAF2 and BAR but not CIDE-B and DR4. Futhermore, our results showed that the treatment of Hep G2 cells with 25 and $50\; {\mu}g/ml$ of RVLEE suppressed PMA-induced COX-2 gene activity. These data suggest that RVLEE have inhibitory effect of cell proliferation, induction of apoptosis and, thus, may offer therapeutic potential in Hep G2.

Effect of Arresting MCF-7 Human Breast Carcinoma Cell at G2/M Phase of Trichosanthes Kirilowii (천화분이 MCF-7 유방암 세포주의 G2/M 세포주기 억제에 미치는 영향)

  • Jeong, Seung-Min;Jeong, Mi-Kyung;Ko, Seong-Gyu;Choi, You-Kyung;Park, Jong-Hyeong;Jun, Chan-Yong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.5
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    • pp.857-862
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    • 2011
  • The purpose of this study is to investigate the anti-proliferative mechanism by Trichosanthes kirilowii (TCK) in MCF-7 human breast carcinoma cell. In this study, we used human breast cancer cell line, Michigan cancer foundation-7 cells (MCF-7 cells). They were co-incubated with 30~200 ${\mu}g$/ml TCK for 48 hours, and cell viability was measured by Water-soluble tetrazolium salt-1 (WST-1) assay. After MCF-7 cells were exposed to 60 ${\mu}g$/ml of TCK for 0, 3, 6, 12, 24, 48 hours, We performed flow analysis cytometry sorting(FACS) and western blot analysis. We investigated the effect of dose-dependent cell growth inhibition by TCK, which could be proved by WST-1 assay. Also, flow cytometry analysis showed that TCK increased percentage of subG1 phase and G2/M phase cell cycle. In addition, TCK induced apoptosis through the expression of caspase-9, -3 and poly(ADP-ribose) polymerase(PARP) activation. Moreover, we showed that ATM-dependent G2/M phase arrest by DNA damage and phosphorylation of chk2, cdc25C, cdc2(Tyr15). Taken together, these results suggest that by G2/M phase arrest through DNA damage and inducing of apoptosis through intrinsic pathway, TCK may have potential tumor suppressor in breast cancer.

Isolation and Morphological Identification of Fresh Water Green Algae from Organic Farming Habitats in Korea (유기농업 생태계로부터 담수 녹조류 분리 및 형태적 동정)

  • Kim, Min-Jeong;Shim, Chang-Ki;Kim, Yong-Ki;Hong, Sung-Jun;Park, Jong-Ho;Han, Eun-Jung;Jee, Hyeong-Jin;Yun, Jong-Chul;Kim, Suk-Chul
    • Korean Journal of Organic Agriculture
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    • v.22 no.4
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    • pp.743-760
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    • 2014
  • This study aimed to isolate and identify freshwater algae from the organic agricultural ecosystems and investigate its biological characteristics to study the possibility of utilizing a biomass freshwater algae in organic farming. In the survey area, average water temperature was $12.4{\sim}28.2^{\circ}C$ and the pH ranges were from 6.1 to 8.5. The solid culture method is more suitable than liquid culture method for isolation of freshwater algae with lower contamination level and higher isolation frequency. A total of 115 strains were isolated from six freshwater algae habitats in nine regions in Korea. BGMM (BG11 Modified Medium) amended with NaNO3 and $KNO_3$ as a nitrogen, and $Na_2CO_3$ as carbon source was designed to isolate and culture freshwater algae. Absorbance of freshwater algae culture has increased dramatically to four days and decreased after eight days after inoculation. CHK008 of the seven isolates showed the highest absorbance in seven days after culturing in BGMM. The optimal pH of BGMM for culturing freshwater algae was pH 6-7. As light intensity increased, growth of freshwater algae increased. Among the five kinds of carbon sources, glucose and galactose promoted good growth of freshwater algae in BGMM. The colony color of purified 16 green algae isolates showed a separation of green, dark and light green, and of them, eleven algae strains showed a strong fluorescent light under fluorescence microscopy. Cell size of the green algae showed a wide range of variation depending on the species. General morphology of the green algae strains was spherical. Chlamydomonas sp. was elliptical, and Chlorella sorokiniana was ellipsoidal and cylindrical. All strains of the green algae except for Chlamydomonas sp. did not have flagella. One isolate of Chlamydomonas sp. and five isolates of C. sorokiniana secreted mucus. Sixteen isolates of 16 green algae were identified as two family and six species, Chlorella vulgalis, C. sorokiniana, C. pyrenoidosa, C. kessleri, C. emersonii, and Chlamydomonas sp. based on their morphological characteristics.

Study on the Fuzzy Inference System for Objectivity of Ground Evaluation in Tunnelling (터널지반 평가의 객관화를 위한 퍼지추론시스템 연구)

  • 조만섭;김영석
    • Tunnel and Underground Space
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    • v.13 no.1
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    • pp.6-19
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    • 2003
  • This study has for its object to increase an objectivity of the observation result in the face mapping of tunnel and to suggest the reasonable support and reinforcement methods to be considered the rock properties. It was developed in this study to the tunnel stability evaluation system(Prototype NFEST) to be used fuzzy set theory and neuro-fuzzy techniques, and this system was verified according to the reliability evaluation between the 36 learning data and the inferred results. When it summarized the results; (1) 12 evaluation items and ranges were proposed to be modified basis on the RMR which are well known to the domestic workers. (2) It was shown that correlation coefficient(│R│) between $RMR_{inf}$ inferred by 12 items and $RMR_{org}$ due to arithmetic total, $RMR_{chk}$ due to subjective judgement of observer are relatively high relationship with each 0.83 and 0.79. (3) Inferred result of the total tunnel safety shows also a good relationship with $RMR_{inf}$ (│R│=0.7) and the rock weathering(│R│=0.84).

Sirt1 Promotes DNA Damage Repair and Cellular Survival

  • Song, Seung-Hyun;Lee, Mi-Ok;Lee, Ji-Seon;Oh, Je-Sok;Cho, Sung-Uk;Cha, Hyuk-Jin
    • Biomolecules & Therapeutics
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    • v.19 no.3
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    • pp.282-287
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    • 2011
  • Sirt1, a nicotinamide adenine dinucleotide ($NAD^+$)-dependent histone deacetylase, is known to deacetylate a number of proteins that are involved in various cellular pathways such as the stress response, apoptosis and cell growth. Modulation of the stress response by Sirtuin 1 (Sirt1) is achieved by the deacetylation of key proteins in a cellular pathway, and leads to a delay in the onset of cancer or aging. In particular, Sirt1 is known to play an important role in maintaining genomic stability, which may be strongly associated with a protective effect during tumorigenesis and during the onset of aging. In these studies, Sirt1 was generated in stably expressing cells and during the stimulation of DNA damage to examine whether it promotes survival. Sirt1 expressing cells facilitated the repair of DNA damage induced by either ionizing radiation (IR) or bleomycin (BLM) treatment. Fastened damaged DNA repair in Sirt1 expressing cells corresponded to prompt activation of Chk2 and ${\gamma}$-H2AX foci formation and promoted survival. Inhibition of Sirt1 enzymatic activity by a chemical inhibitor, nicotinamide (NIC), delayed DNA damage repair, indicating that promoted DNA damage repair by Sirt1 functions to induce survival when DNA damage occurs.

Knockdown of UHRF1 by Lentivirus-mediated shRNA Inhibits Ovarian Cancer Cell Growth

  • Yan, Feng;Shao, Li-Jia;Hu, Xiao-Ya
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.4
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    • pp.1343-1348
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    • 2015
  • Human UHRF1 (ubiquitin-like PHD and RING finger domain-containing 1) has been reported to be over-expressed in many cancers, but its role in ovarian cancer remains elusive. Here, we determined whether knockdown of UHRF1 by lentivirus-mediated shRNA could inhibit ovarian cancer cell growth. Lentivirus-mediated short hairpin RNAs (lv-shRNAs-UHRF1) were designed to trigger the gene silencing RNA interference (RNAi) pathway. The efficiency of lentivirus-mediated shRNA infection into HO-8910 and HO-8910 PM cells was determined using fluorescence microscopy to observe lentivirus-mediated GFP expression and was confirmed to be over 80 percent. UHRF1 expression in infected HO-8910 and HO-8910 PM was evaluated by real-time PCR and Western blot analysis. The Cell Counting Kit-8 (CCK-8) assay was used to measure cell viability; flow cytometry and Hoechst 33342 assay was applied to measure cell cycle arrest and apoptosis. Cell invasion was assessed using transwell chambers. Our results demonstrated that the loss of UHRF1 promoted HO-8910 and HO-8910 PM cell apoptosis, while inhibiting cell proliferation. In addition, UHRF1 knockdown significantly inhibited the invasion of human ovarian cancer cells. In the present study, we also showed that depleting HO-8910 cells of UHRF1 caused activation of the DNA damage response pathway, with the cell cycle arrested in G2/M-phase. The DNA damage response in cells depleted of UHRF1 was illustrated by phosphorylation of CHK (checkpoint kinase) 2 on Thr68, phosphorylation of CDC25 (cell division control 25) on Ser 216 and phosphorylation of CDK1 (cyclin-dependent kinase 1) on Tyr 15.

Protective Effects of Nypa fruticans Wurmb against Oxidative DNA Damage and UVB-induced DNA Damage

  • So-Yeon Han;Tae-Won Jang;Da-Yoon Lee;Seo-Yoon Park;Woo-Jin Oh;Se Chul Hong;Jae-Ho Park
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2023.04a
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    • pp.54-54
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    • 2023
  • Nypa fruticans Wurmb (N. fruticans) is a plant that belongs to Araceae and N. fruticans is mainly found in tropical mangrove systems. The parts (leaves, stems, and roots) of N. fruticans are traditionally used for asthma, sore throat, and liver disease. N. fruticans contains flavonoids and polyphenols, which are substances that have inhibitory effects on cancer and oxidant. In previous studies, some pharmaceutical effects of N. fruticans on melanogenesis and inflammation have been reported. The present study is conducted to investigate the effect of the ethyl acetate fraction of N. fruticans (ENF) on oxidative DNA damage and UVB-induced DNA damage. DNA damage response (DDR) pathway is important in research on cancer, apoptosis, and so on. DDR pathways are considered a crucial factor affecting the alleviation of cellular damage. ENF could reduce oxidative DNA damage derived from reactive oxygen species by the Fenton reaction. Also, ENF reduced the intensity of intracellular ROS in the live cell image by DCFDA assay. UVB is known to cause skin and cellular damage, then finally contribute to causing the formation of tumors. As for the strategies of reducing DNA damage by UVB, inhibition of p53, H2AX, and Chk2 can be important indexes to protect the human body from DNA damage. As a result of confirming the protective effect of ENF for UVB damage, MMPs significantly decreased, and the expression of apoptosis-related factors tended to decrease. In conclusion, ENF can provide protective effects against double-stranded DNA break (DSB) caused by oxidative DNA damage and UVB-induced DNA damage. These results are considered to be closely related to the protective effect against radicals based on catechin, epicatechin, and isoquercitrin contained in ENF. Based on these results, it is thought that additional mechanism studies for inhibiting cell damage are needed.

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ATM Signaling Pathway Is Implicated in the SMYD3-mediated Proliferation and Migration of Gastric Cancer Cells

  • Wang, Lei;Wang, Qiu-Tong;Liu, Yu-Peng;Dong, Qing-Qing;Hu, Hai-Jie;Miao, Zhi;Li, Shuang;Liu, Yong;Zhou, Hao;Zhang, Tong-Cun;Ma, Wen-Jian;Luo, Xue-Gang
    • Journal of Gastric Cancer
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    • v.17 no.4
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    • pp.295-305
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    • 2017
  • Purpose: We previously found that the histone methyltransferase suppressor of variegation, enhancer of zeste, trithorax and myeloid-nervy-deformed epidermal autoregulatory factor-1 domain-containing protein 3 (SMYD3) is a potential independent predictive factor or prognostic factor for overall survival in gastric cancer patients, but its roles seem to differ from those in other cancers. Therefore, in this study, the detailed functions of SMYD3 in cell proliferation and migration in gastric cancer were examined. Materials and Methods: SMYD3 was overexpressed or suppressed by transfection with an expression plasmid or siRNA, and a wound healing migration assay and Transwell assay were performed to detect the migration and invasion ability of gastric cancer cells. Additionally, an MTT assay and clonogenic assay were performed to evaluate cell proliferation, and a cell cycle analysis was performed by propidium iodide staining. Furthermore, the expression of genes implicated in the ataxia telangiectasia mutated (ATM) pathway and proteins involved in cell cycle regulation were detected by polymerase chain reaction and western blot analyses. Results: Compared with control cells, gastric cancer cells transfected with si-SMYD3 showed lower migration and invasion abilities (P<0.05), and the absence of SMYD3 halted cells in G2/M phase and activated the ATM pathway. Furthermore, the opposite patterns were observed when SMYD3 was elevated in normal gastric cells. Conclusions: To the best of our knowledge, this study provides the first evidence that the absence of SMYD3 could inhibit the migration, invasion, and proliferation of gastric cancer cells and halt cells in G2/M phase via the ATM-CHK2/p53-Cdc25C pathway. These findings indicated that SMYD3 plays crucial roles in the proliferation, migration, and invasion of gastric cancer cells and may be a useful therapeutic target in human gastric carcinomas.