• Title/Summary/Keyword: Blotting, western

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The Effect of Inhibition of Heme Oxygenase-1 on Chemosensitivity of Cisplatin in Lung Cancer Cells (폐암세포주에서 Heme Oxygenase-1의 억제가 Cisplatin의 항암제 감수성에 미치는 영향)

  • Kim, So-Young;Kim, Eun-Jung;Jang, Hye-Yeon;Hwang, Ki-Eun;Park, Jung-Hyun;Kim, Hwi-Jung;Jo, Hyang-Jeong;Yang, Sei-Hoon;Jeong, Eun-Taik;Kim, Hak-Ryul
    • Tuberculosis and Respiratory Diseases
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    • v.62 no.1
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    • pp.33-42
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    • 2007
  • Background: Heme oxygenase-1 (HO-1) is known to modulates the cellular functions, including cell proliferation and apoptosis. It is known that a high level of HO-1 expression is found in many tumors, and HO-1 plays an important role in rapid tumor growth on account of its antioxidant and antiapoptotic effects. Cisplatin is a widely used anti-cancer agent for the treatment of lung cancer. However, the development of resistance to cisplatin is a major obstacle to its use in clinical treatment. We previously demonstrated that inhibiting HO-1 expression through the transcriptional activation of Nrf2 induces apoptosis in A549 cells. The aim of this study was to determine of the inhibiting HO-1 enhance the chemosensitivity of A549 cells to cisplatin. Materials and Methods: The human lung cancer cell line, A549, was treated cisplatin, and the cell viability was measured by a MTT assay. The change in HO-1, Nrf2, and MAPK expression after the cisplatin treatment was examined by Western blotting. HO-1 inhibition was suppressed by ZnPP, which is a specific pharmacologic inhibitor of HO activity, and small interfering RNA (siRNA). Flow cytometry analysis and Western blot were performed in to determine the level of apoptosis. The level of hydrogen peroxide ($H_2O_2$) generation was monitored fluoimetrically using 2',7'-dichlorofluorescein diacetate. Results: The A549 cells showed more resistance to the cisplatin treatment than the other cell lines examined, whereas cisplatin increased the expression of HO-1 and Nrf2, as well as the phosphorylation of MAPK in a time-dependent fashion. Inhibitors of the MAPK pathway blocked the induction of HO-1 and Nrf2 by the cisplatin treatment in A549 cells. In addition, the cisplatin-treated A549 cells transfected with dither the HO-1 small interfering RNA (siRNA) or ZnPP, specific HO-1 inhibitor, showed in a more significantly decrease in viability than the cisplatin-only-treated group. The combination treatment of ZnPP and cisplatin caused in a marked increase in the ROS generation and a decrease in the HO-1 expression. Conclusion: Cisplatin increases the expression of HO-1, probably through the MAPK-Nrf2 pathway, and the inhibition of HO-1 enhances the chemosensitivity of A549 cells to cisplatin.

Presence of Carbonic Anhydrase III in Liver of Flounder, Limanda yokohamae

  • Kho, Kang-Hee;Choi, Kap-Seong
    • Food Science and Biotechnology
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    • v.14 no.4
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    • pp.551-553
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    • 2005
  • Carbonic anhydrase III was found in liver of flounder, Limanda yokohamae. Protein was isolated from cytosolic extracts and identified using SDS-PAGE and isolectric focusing. Specific protein bands with molecular weight of 30 kDa and pIs of 7.0 and 6.5 were detected by Western blotting. This is the first report of identification of carbonic anhydrase III from L. yokohamae.

Characterization and Localization of the Murine nm23-M5 in Mouse Testis

  • Kang, Sung-Jo;Park, Yun-Jung;Kim, Jin-Hoi
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.223-223
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    • 2004
  • Nucleoside diphosphate kinases(NDPKs) are ubiquitous enzymes involved in numerous regulatory processes associated with transcriptional regulation, cell proliferation, development, and differentiation. In this study, we was examined characterization and localization of the nm23-M5 in mouse testis by Western blotting, immunohistochemical and conforcal imaging study using specific antibodies raised against nm23-M5. (omitted)

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Allicin-induced apoptosis of gastric epithelial cells is associated with changes of caspase-independent effector and involvement of PKA

  • Baeg, Hye-Kyoung;Rhee, Dong-Kwon;Pho, Suhk-Neung
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.166.2-166.2
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    • 2003
  • Garlic (Allium sativum) has been used as a general food and a remedy in Oriental for a long time. Since garlic compounds have been also shown to inhibit growth of tumors and to modulate the activity of carcinogenesis, the effects of allicin on growth and survival in human gastric epithelial cells were evaluated by cell viability, cell cycle analysis and DNA fragmentation. Protein levels of cytochrome C, Bcl-xL, Bax and AIF were detected by Western blotting. Effects of recombinant VacA on caspase proteases activity were also determined. (omitted)

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Inhibition of VRK1 suppresses proliferation and migration of vascular smooth muscle cells and intima hyperplasia after injury via mTORC1/β-catenin axis

  • Sun, Xiongshan;Zhao, Weiwei;Wang, Qiang;Zhao, Jiaqi;Yang, Dachun;Yang, Yongjian
    • BMB Reports
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    • v.55 no.5
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    • pp.244-249
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    • 2022
  • Characterized by abnormal proliferation and migration of vascular smooth muscle cells (VSMCs), neointima hyperplasia is a hallmark of vascular restenosis after percutaneous vascular interventions. Vaccinia-related kinase 1 (VRK1) is a stress adaption-associated ser/thr protein kinase that can induce the proliferation of various types of cells. However, the role of VRK1 in the proliferation and migration of VSMCs and neointima hyperplasia after vascular injury remains unknown. We observed increased expression of VRK1 in VSMCs subjected to platelet-derived growth factor (PDGF)-BB by western blotting. Silencing VRK1 by shVrk1 reduced the number of Ki-67-positive VSMCs and attenuated the migration of VSMCs. Mechanistically, we found that relative expression levels of β-catenin and effectors of mTOR complex 1 (mTORC1) such as phospho (p)-mammalian target of rapamycin (mTOR), p-S6, and p-4EBP1 were decreased after silencing VRK1. Restoration of β-catenin expression by SKL2001 and re-activation of mTORC1 by Tuberous sclerosis 1 siRNA (siTsc1) both abolished shVrk1-mediated inhibitory effect on VSMC proliferation and migration. siTsc1 also rescued the reduced expression of β-catenin caused by VRK1 inhibition. Furthermore, mTORC1 re-activation failed to recover the attenuated proliferation and migration of VSMC resulting from shVrk1 after silencing β-catenin. We also found that the vascular expression of VRK1 was increased after injury. VRK1 inactivation in vivo inhibited vascular injury-induced neointima hyperplasia in a β-catenin-dependent manner. These results demonstrate that inhibition of VRK1 can suppress the proliferation and migration of VSMC and neointima hyperplasia after vascular injury via mTORC1/β-catenin pathway.

Pectolinarigenin ameliorated airway inflammation and airway remodeling to exhibit antitussive effect

  • Quan He;Weihua Liu;Xiaomei Ma;Hongxiu Li;Weiqi Feng;Xuzhi Lu;Ying Li;Zi Chen
    • The Korean Journal of Physiology and Pharmacology
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    • v.28 no.3
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    • pp.229-237
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    • 2024
  • Cough is a common symptom of several respiratory diseases. However, frequent coughing from acute to chronic often causes great pain to patients. It may turn into cough variant asthma, which seriously affects people's quality of life. For cough treatment, it is dominated by over-the-counter antitussive drugs, such as asmeton, but most currently available antitussive drugs have serious side effects. Thus, there is a great need for the development of new drugs with potent cough suppressant. BALB/c mice were used to construct mice model with cough to investigate the pharmacological effects of pectolinarigenin (PEC). Hematoxylin-eosin and Masson staining were used to assess lung injury and airway remodeling, and ELISA was used to assess the level of inflammatory factor release. In addition, inflammatory cell counts were measured to assess airway inflammation. Airway hyperresponsiveness assay was used to assess respiratory resistance in mice. Finally, we used Western blotting to explore the potential mechanisms of PEC. We found that PEC could alleviate lung tissue injury and reduce the release of inflammatory factors, inhibit of cough frequency and airway wall collagen deposition in mice model with cough. Meanwhile, PEC inhibited the Ras/ERK/c-Fos pathway to exhibit antitussive effect. Therefore, PEC may be a potential drug for cough suppression.

Usefulness of 32kDa Polypeptide Detection of Theileria sergenti in Monitoring Treatment Progress of Bovine Theileriosis (소의 테이레리아병 치료시 Theileria sergenti의 32kDa Polypeptide 검출의 유용성)

  • Kim, Byeong-Soo
    • Korean Journal of Veterinary Research
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    • v.42 no.3
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    • pp.377-382
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    • 2002
  • Bovine piroplasmosis caused by Theileria sergenti is a major cause of economic loss in livestock industry. Five cattle infected with Theileria sergenti showing severe and fatal anemia, confirmed by indirect immunofluorescent assay(IFA), were used in this study. Four cattle were treated with diminazene aceturate and one was not treated as the control. The therapeutic effect of diminazene aceturate against Theileria sergenti infection was monitored by detecting the 32kDa polypeptide specific for Theileria sergenti by the western blotting with both polyclonal and monoclonal antibodies. The 32kDa polypeptide detected at the beginning of diminazene aceturate treatment was not detectable after the treatment. It is postulated that the detection of the 32kDa polypeptide specific for Theileria sergenti may be a good tool for the diagnosis and monitoring the treatment progress of Theileria sergenti infection.

Induction of Porcine Oocytes Maturation by MAP Kinase (MAP Kinase에 의한 돼지 난성숙의 유기)

  • 장규태;박미령;윤창현
    • Korean Journal of Animal Reproduction
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    • v.22 no.3
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    • pp.277-286
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    • 1998
  • The effect of MAP kinase activity on maturation of porcine oocytes was investigated. MAP kinase was detected by immunofluorescence staining in nucleus of oocytes just before entering GVBD (germinal vesicle breakdown)stage. In a Western blot with GV (germinal vesicle) from these oocytes (cultured for 25 hours), a shift of MAP kinase band a, pp.ared, suggesting an activated stage of the kinase. No activity was shown in the blot with GV isolated from, oocytes cultured for 0 hour. To confirm that activation of MAP kinase induce GVBD, we microinjected MAP kinase purified from matured oocytes starfish into the cytoplasm of oocytes in GV stage (cultured for 0 hour). The injected MAP kinase did not cause early a, pp.arance of GVBD. No oocytes showed GVBD state until 20 hours of culture. Activity of MAP kinase did not increase significantly after the injection. When the exogenous MAP kinase was injected into GV, GVBD was induced in about 20% of oocytes cultured for 5 to 10 hours. These results suggest that MAP kinase is traslocated to nucleus and function as a factor inducing GVBD.

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Antitumorigenic Effects of Tannin From Persimmon Leaves on Sarcoma 180-induced Tumor in Mice

  • Moon, Sung-Chai;Park, Kyong-Hee;Rhew, Tae-Hyong;Park, Kun-Young;Kim, Byeong-Gee
    • Preventive Nutrition and Food Science
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    • v.3 no.1
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    • pp.92-97
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    • 1998
  • The changes of morphology and protein pattern of sarcoma 180 cells treated with or without trannins extracted from persimmon leaves were evaluated by light microscopy, electrophoresis and Western blotting. The sarcoma 180 cells treated with tannins increased the amount of proteins which presumably were intermediate filament cytokeratins detected by electrophoresis and Western blot. Tannins was indirectly cytotoxic to the sarcoma 180 cells and increased the intermediate filament protein level in the cells.

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Enzyme-linked immunosorbent assay for detection of Trichinella spiralis antibodies and the surveillance of selected pig breeding farms in the Republic of Korea

  • Wee, Sung-Hwan;Lee, Chung-Gil;Joo, Hoo-Don;Kang, Yung-Bai
    • Parasites, Hosts and Diseases
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    • v.39 no.3
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    • pp.261-264
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    • 2001
  • Trichinellosis is a parasitic zoonosis of public health importance. It is caused by Trichinella spiralis which has a wide host range including humans. In the present communication, the ELISA technique was employed on a total of 803 blood samples from 7 selected pig breeding farms in 1996 for diagnosis and surveillance of trichinellosis. Out of the entire 803 samples, nine were found to be suspected while one was positive by ELISA. But western blot analyses employed for further confirmation have shown that all of 10 samples did not react to larval excretory-secretory product antigens. These results indicate that pig breeding farms included in the present study are free from trichinellosis . However, it does not mean Korea is free from trichinellosis since human trichinellosis has recently been reported. The necessity of continued surveillance for trichinellosis in both pigs and wild animals was discussed.

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