• Title, Summary, Keyword: 세포예정사

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대기압 플라즈마 젯의 산소 첨가량에 따른 세포 예정사 효과

  • Jo, Hye-Min;Jeong, Tae-Hun;Bae, Se-Hwan;Im, Seon-Hui
    • Proceedings of the Korean Vacuum Society Conference
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    • pp.452-452
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    • 2012
  • 저온 플라즈마를 발생시키는 대기압 마이크로-플라즈마 젯(Micro-plasma jet)을 이용하여 플라즈마와 세포와의 상호작용에 대한 연구를 진행하였다. 실험에 사용된 세포는 인체의 방광암 세포(Human bladder carcinoma cell, EJ)이며, 플라즈마 처리 후 bioassay를 통하여 세포 예정사 효과를 확인하였다. 수십 kHz (low frequency)의 펄스파 전압을 인가하여 발생시킨 플라즈마는 형성 기체로 헬륨을 사용하였고, 광 방출 분광법으로 산소의 첨가량에 따른 활성 종들의 변화를 비교해 보았다. 플라즈마 처리 후에는 DAPI staining을 통하여 세포 예정사에서 형성되는 apoptotic body를 확인하였고, 세포막 외부로 이동하는 Phosphatidic Serin (PS)과 결합하는 Annexin-V assay를 통하여 apoptosis rate를 측정하였다. 이를 바탕으로 암세포에 미치는 플라즈마 활성종의 영향을 분석하였다.

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Cooperative stimulation of cisplatin-mediated apoptosis by hepatitis B virus X Protein and hepatitis C virus core Protein (B형 간염 바이러스 X 단백질과 C형 간염 바이러스의 코어 단백질에 의한 cisplatin-매개성 세포 예정사의 협조적 촉진)

  • Kwun, Hyun-Jin;Jang, Kyung-Lib
    • Journal of Life Science
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    • v.17 no.6
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    • pp.766-771
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    • 2007
  • The co-infection with hepatitis B virus (HBV) and hepatitis C Virus (HCV) is associated with a more severe liver disease and increased frequency in the development of hepatocellular carcinoma com-pared to those with single infection. Here, we demonstrated that HBV X protein (HBx) and HCV Core cooperatively up-regulated the level of p53 in human hepatoma HepG2 cells. The elevated p53 subsequently stimulated the expression of proapoptotic Bax whereas it repressed the expression of antiapoptotic Bcl2. These effects, however, were not observed in p53-negative Hep3B cells. Consistently to their cooperative regulation of apoptotic effectors, HBx and HCV Core additively stimulated cisplatin-mediated apoptotic cell death of HepG2 but not of Hep3B cells. These results may help to explain the development of a more severe liver disease in patients co-infection with HBV and HCV as well as some contradictory results on the roles of HBx and Core in apoptosis.

Ultrastructural study of mouse ovary under X-ray irradiation (방사선 조사선량에 따른 생쥐 난소의 미세구조적 연구)

  • Yoon, Chul-Ho
    • Journal of radiological science and technology
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    • v.28 no.3
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    • pp.249-254
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    • 2005
  • This study investigated the structural changes of folliculus ovarii according to the dose of the X-rays when mice were exposed to X-rays from 6MeV LINAC. The minute structural changes of folliculus ovarii were observed through an electron microscope with high magnification. Nuclei and protoplasm of granular cells in growing folliculus ovarii abruptly underwent minute structural changes according to the increase of dose of X-rays. Cell residue, by-product of cell decease, neutrophil and macrophage around follicular antrum were observed. The minute structural changes in granular cells showed typical characteristics of apoptosis: the increase of electronic density due to nuclear condensation, fragmentation of nuclei, and atrophy of protoplasm. Necrosis of cells was identified, but it was not so remarkable. Macrophage scattered with apoptotic bodies.

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Potential Involvement of p62, a Phosphotyrosine-independent Ligand of SH2 Domain of $p56^{lck}$, on UV-induced Apoptosis in Jurkat T-cell Line ($p56^{lck}$ SH2 domain 결합 단백질 p62가 Jurkat T-세포주의 세포예정사에 미치는 영향)

  • 정인실
    • Development and Reproduction
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    • v.2 no.2
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    • pp.165-171
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    • 1998
  • p62 is a novel cytoplsmic protein that binds to SH2 domain of p56$^{lck}$, lymphocyte-specific protein tyrosine kinase, and the expression of p62 was observed in most tissues. In addition p62 interacts with various proteins including ubiquitin and atypical PKC isoform, indicating its diverse biological role in different tissues. However, little is known about functional connection between p62 and its binding proteins. In the present study, a novel cellular protein, p62 has been shown to bind to 14-3-3 $\tau$ isoform that is specific for T cells. Moreover, overexpression of p62 in T cells caused to delay onset of UV-induced apoptosis characterized by DNA fragmentation and breakdown of poly (ADP-ribose) polymerase (PARP). Lately, 14-3-3 proteins have been shown to mediate survival signal via interacting proapoptotic Bad protein in the Iymphocyte. These results suggested the presence of p62-mediated regulatory mechanism during apoptosis in T cells, in which activation-induced apoptotic signal could be interfered by p62 and 14-3-3 protein.n.

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Morphological Changes of Mouse Ovary by X-Ray Irradiation (방사선 조사선량에 따른 생쥐 난소의 형태학적 변화)

  • Yoon, Chul-Ho;Choi, Jong-Woon;Yoon, Surk-Hwan
    • Journal of Radiation Protection and Research
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    • v.32 no.4
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    • pp.140-156
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    • 2007
  • This research was performed to investigate the morphological changes of folliculus ovary according to the radiation dose. The whole body radiation of 200 cGy, 400 cGy, and 600 cGy was given to the each groups of 5 months-aged female mouse. Various staining methods used in this research are: Hematosylin-Eosin method, and immunohistochemistrical methods using BrdU, TUNEL, p53, p21, PCNA and inhibin. The minute structural changes of folliculus ovary were observed through an electron microscope with high magnification. The morphological changes of growing folliculus ovary became distinct as the dose of X-rays increased. Especially, the nuclei of granular cells showed manifest condensation and the changes of the transparent zone were distinct. As a result of histochemical reaction according to Masson's trichrome method and reticular fiber method, the changed granular cells, the deformed basilar membrane of folliculus ovary and the abnormal arrangement of the reticular fiber were observed. In the reaction of BrdU, the granular cells of normal folliculus ovary with positive reaction rapidly decreased according to the increase of the dose of X-rays. In TUNEL study, granular cells showing positive reaction in retarded folliculus ovary were expanded to growing folliculus ovary and primordial folliculus ovary according to the increase of the dose of X-rays. In case of 600 cGy of X-rays, oocyte underwent apoptosis. In p53 immunohistochemistry, p53 manifested to be stronger as the dose of X-rays increased. p53 reactivity was manifested distinctively in all cells comprising folliculus ovary following irradiation of 600 cGy. p21 was manifested in granular cells of folliculus ovary and showed very positive reaction around follicular antrum according to the increase of the dose of X-rays. In PCNA, positive reaction was manifested in growing folliculus ovary, mature folliculus ovary and primordial folliculus ovary, but the extent of the reaction decreased as the dose of the X-rays decreased. The finding that the reaction of granular cells around folliculus ovary was stronger than that near follicular membrane indicates that what was damaged first by X-ray was the cells near folliculus ovary and follicular antrum. The reactivity of $inhibin-{\alpha}$ showed difference according to the growing stage of folliculus ovary: $inhibin-{\alpha}$ showed the most strong reaction in mature folliculus ovary with follicular antrum. There was strong reaction in granular cells around follicular membrane but $inhibin-{\alpha}$ did not occur at all in theca cells comprising follicular membrane. $Inhibin-{\alpha}$ in ovary tissue exposed to 400 cGy of X-rays was manifested more strongly than in ovary tissue exposed to 600 cGy of X-rays, which was related to the phenomenon that granular cells of mature folliculus ovary underwent necrosis or apoptosis increasingly due to X-rays. In an electron microscope with high magnification, nuclei and protoplasm of granular cells in growing folliculus ovary abruptly underwent minute structural changes according to the increase of dose of X-rays. Cell residue, by-product of cell decease, neutrophil and macrophage around follicular antrum were observed. The minute structural changes in granular cells showed typical characteristics of apoptosis: the increase of electronic density due to nuclear condensation, fragmentation of nuclei and atrophy of protoplasm. Necrosis of cells was identified but it was not so remarkable. Macrophage with apoptotic bodies was scattered. Proportional to the radiation dose, we found that the generation of heterogeneous substance of normal ovary texture's follicular fluid, the emergence of dyeing characteristic in the basilar membrane of folicle, the generation of apoptosis, and the transformation of macrophages, etc. From this results, we can infer the possible radiation hazard on the ovary of cervix cancer patient with radiation therapy.

다양한 기체를 사용한 대기압 플라즈마 젯에 대한 세포 내 활성 산소종의 영향 연구

  • Jo, Hye-Min;Kim, Seon-Ja;Jeong, Tae-Hun;Im, Seon-Hui
    • Proceedings of the Korean Vacuum Society Conference
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    • pp.542-542
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    • 2013
  • 저온 플라즈마를 발생시키는 대기압 마이크로-플라즈마 젯(Micro-plasma jet)을 이용하여 플라즈마와 세포와의 상호작용에 대한 연구를 진행하였다. 세포의 대사과정에서 생성되는 활성산소 종(Reactive Oxygen Species, ROS)은 세포에 산화 스트레스를 유발시킨다. 이러한 스트레스는 세포 예정사(programmed cell death)의 원인이 된다. 플라즈마 형성 기체로 헬륨, 아르곤, 질소를 사용하여 각각의 기체에 따른 세포의 형태 변화 및 세포 내 활성 산소 종의 영향을 분석하였다. 실험에 사용된 세포는 인체의 폐암 세포[Human lung cancer cell, A549]이며 플라즈마 처리 후 Intracellular ROS assay를 통하여 플라즈마에서 발생되는 활성 산소 종(Reactive Oxygen Species, ROS)이 세포 내에 들어가 활성 산소 종을 증가시키는 것을 확인하였다. 이때, 플라즈마에서 발생되는 활성 산소 종(Reactive Oxygen Species, ROS)들은 광 방출 스펙트럼(Optical Emission Spectroscopy)로 분석하였고, 기체별로 비교하여 보았다. 또한, 이 때 발생되는 플라즈마의 전류-전압 특성에 따른 optical intensity를 비교하였다.

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Study of Histochemical Changes of Mouse Ovary Under X-ray Irradiation (방사선 조사선량에 따른 생쥐 난소의 조직화학적 변화)

  • Yoon, Chul-Ho
    • Journal of radiological science and technology
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    • v.29 no.4
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    • pp.275-284
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    • 2006
  • This research investigated the morphological changes of folliculus ovary according to the dose of the X-rays when mice were exposed to X-rays, in terms of histochemical reaction. Dyeing methods such as Masson's trichrome method and reticular fiber method were used. The morphological changes of growing folliculus ovary got distinct as the dose of X-rays increased. Especially, the nuclei of granular cells showed manifest condensation and the changes of the transparent zone were distinct. As a result of histochemical reaction according to Masson's trichrome method and reticular fiber method, the changed granular cells, the deformed basilar membrane of folliculus ovary and the abnormal arrangement of the reticular fiber were observed.

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대기압 플라즈마 젯의 질소종에 의한 방출광 및 온도변화 연구

  • Park, Sang-Hu;Jeong, Hui-Su;Kim, Seon-Ja;Choe, Won-Ho
    • Proceedings of the Korean Vacuum Society Conference
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    • pp.345-345
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    • 2011
  • 최근 대기압 플라즈마 젯을 이용한 바이오/메디컬의 활발한 응용연구가 진행 중이다. 박테리아 및 세균의 살균은 물론 암세포 세포예정사에 핵심적인 역할을 하는 활성산소종(Reactive Oxygen Species, ROS) 또는 다양한 라디칼들은 대기압 플라즈마의 다양한 변수를 이용하여 조절할 수 있다고 알려져 있다. 수십 kHz의 고전압에서 발생된 마이크로 헬륨 플라즈마 젯에서 질소종의 제어를 통해 같은 부피의 플라즈마 젯에서의 방출광을 살펴보았다. 또한 광섬유센서를 이용하여 플라즈마의 기체온도를 측정하고 Boltzmann plot method를 통해 전자의 여기온도 변화를 관찰하였다. 실험의 결과, 같은 부피의 플라즈마에서 질소종이 증가할 때 기체온도는 큰 변함이 없지만 여기온도가 증가하는 것을 관찰하였다. 시간분해 이미지 촬영으로 질소종의 양에 따른 플라즈마 불릿의 속도 변화를 분석을 하였고, 최종적으로 대기압 플라즈마 젯의 질소종 변화에 따른 대장균의 비활성화 정도를 관찰하였다.

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Putative Bax inhibitor from rice a conserved cell death suppressor, is isolated by yeast functional screening (효모 기능 선발을 이용한 벼의 세포사유발을 억제하는 유전자 선발)

  • Lee, Gyu Ho;Son, Ye Jin;Sawitri, Widhi Diya;Sohn, Jae-Keu;Kim, Kyung-Min
    • Current Research on Agriculture and Life Sciences
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    • v.29
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    • pp.37-42
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    • 2011
  • The plant-homologue of Bax Inhibitor, a gene described to suppress the cell death induced by Bax gene expression in yeast, was isolated from rice (Oryza sativa L.). Nucleic acid sequence and amino acid sequence were 741 bp and 247 bp, respectively. The amino acid sequence of the predicted protein was well conserved in plant (84 % in amino acids) and contained five membrane-spanning segments.

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Relation of Poly(ADP-ribose) Polymerase Cleavage and Apoptosis Induced by Paclitaxel in HeLa S3 Uterine Cancer Cells (HeLa S3 자궁암 세포에서 paclitaxel 에 의해 유도된 Poly(ADP-ribose) Polymerase 분철과 세포자멸사와의 관계)

  • Chang, Jeong-Hyun;Kim, Kwang-Youn;Ahn, Soon-Cheol;Kwon, Heun-Young
    • Journal of Life Science
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    • v.17 no.8
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    • pp.1027-1033
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    • 2007
  • Although paclitaxel induces apoptosis of cancer cells, its exact mechanism of action is not yet known. The present study has been performed to determine whether influence of paclitaxel in HeLa $S_{3}$ uterine cancer cells. Three assays were employed in this study: cell cytotoxicity, morphological assessments of apoptotic cells (DAPI staining assay), and western blot analysis. The results indicated that paclitaxel has cytotoxic effects in HeLa $S_{3}$ cells. Especially, the $IC_{50}$ value of paclitaxel was about 1 ${\mu}M$. And morphological changes (fragmentation) of cells were observed by paclitaxel in HeLa $S_{3}$ cells. The flow cytometric analysis of paclitaxel-treated cells indicated a block of G2/M phase. The results that pacli-taxel regulates the cell cycle, especially Sub-$G_{1}$ phase. Paclitaxel induces apoptosis of HeLa $S_{3}$ cells via PARP-dependent fashion, and this apoptosis is related to disappearance of Bcl-2 proteins.