자궁경부암 세포에서 시스플라틴이 소포체 스트레스에 미치는 영향
THE EFFECT OF CISPLATIN ON ENDOPLASMIC RETICULUM STRESS OF HUMAN CERVICAL CANCER CELLS
2
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
목적항암제 시스플라틴의 세포독성 기전은 활성산소종과 lipid peroxydation의 증가와 DNA adduct의 형성 등이 알려져 있다. 이 연구에서는 시스플라틴이 자궁경부암 세포인 Hela 세포에서 과량의 NO와 세포 내 칼슘 이온의 농도 증가를 통해 소포체 스트레스 관련 단백질들의 발현을 증가시킴을 확인하고자 하였다. 연구방법Hela세포는 시스플라틴으로 처리하였고 다음으로 소포체 스트레스 반응은 western blot, Flow cytometry, fluorescence 분석을 이용하여측정하였다. 결과시스플라틴에 의해 유도되어지는 세포자멸사 현상은 유도성 NO 합성효소의 특이적 억제제인 1400 W와 세포 내 칼슘이온 소거제인 1,2-bis-(o-aminophenoxy)ethane-N,N,N’,N’-tetraacetic acid tetra-(acetoxymethyl) ester에 의해 감소되었다. 이러한 약물들은 또한 미토콘드리아의 막전위를 회복시킴으로써 미토콘드리아에 의한 세포자멸사 신호전달은 억제하였다. 결론시스플라틴에 의한 세포자멸사 현상은 초기 시간대의 칼슘이온 농도의 증가와 그에 따른 미토콘드리아에 의한 세포자멸사 신호전달체계에 의해 유도되며, 이때 소포체가 중요하게 관여되어 있음을 확인할 수 있었다.
Abstract
Objective Cis-diamminedichloroplatinum (cisplatin) is a widely used chemotherapeutic agent. A number of evidences in cytotoxic mechanism of cisplatin, including perturbation of redox status, increase in lipid peroxydation, formation of DNA adduct, have been suggested. The author hypothesized that cisplatin would mediate apoptosis via endoplasmic reticulum (ER) stress in human cervical cancer cell. Methods Human cervical cancer cell line (Hela cells) were treated with cisplatin and then ER stress-related response were performed using western blot, Flow cytometry and fl uorescence analysis. Results After addition of cisplatin to Hela cells, the author observed an expression of ER stress response genes through a gradual increase of nitric oxide and cytosolic Ca2+ concentration. Cisplatin-induced apoptosis can be inhibited by the inducible nitric oxide synthase inhibitor, 1400 W, and intracellular Ca2+ chelator, 1, 2-bis-(o-aminophenoxy)ethane-N,N,N’,N’-tetraacetic acid tetra-(acetoxymethyl)ester (BAPTA-AM). These inhibitors also reduced mitochondrial apoptotic signals, such as mitochondrion membrane potential disruption, cytochrome c release and eventually reduced the death of Hela cells. Conclusion Taken together, ER would seem to contribute to cisplatin-induced apoptosis via both the early release of Ca2+ and the late amplifi cation of mitochondria-mediated apoptotic signals.
참고문헌 (22)
- Kaufman RJ, "Stress signaling from the lumen of the endoplasmic reticulum: coordination of gene transcriptional and translational controls", Genes Dev, 13, pp.1211~1233 (1999)
- Mori K, "Tripartite management of unfolded proteins in the endoplasmic reticulum", Cell, 101, pp.451~454 (2000)
- Baltzis D, "The eIF2alpha kinases PERK and PKR activate glycogen synthase kinase 3 to promote the proteasomal degradation of p53", J Biol Chem, 282, pp.31675~31687 (2007)
- Bobrovnikova-Marjon E, "Coping with stress: ATF6alpha takes the stage", Dev Cell, 13, pp.322~324 (2007)
- Gass JN, "The unfolded protein response of B-lymphocytes: PERK-independent development of antibody-secreting cells", Mol Immunol, 45, pp.1035~1043 (2008)
- Yamamoto K, "Transcriptional induction of mammalian ER quality control proteins is mediated by single or combined action of ATF6alpha and XBP1", Dev Cell, 13, pp.365~376 (2007)
- Endo H, "Activation of insulin-like growth factor signaling induces apoptotic cell death under prolonged hypoxia by enhancing endoplasmic reticulum stress response", Cancer Res, 67, pp.8095~8103 (2007)
- Henshall DC, "Modulators of neuronal cell death in epilepsy", Curr Opin Pharmacol, 8, pp.75~81 (2008)
- Farm RJ, "Cisplatin and platinum analogues: Recent advances", Curr Opin Oncol, 4, pp.1073~1079 (1992)
- Oyadomari S, "Endoplasmic reticulum stressmediated apoptosis in pancreatic beta-cells", Apoptosis, 7, pp.335~345 (2002)
- Harding HP, "PERK is essential for translational regulation and cell survival during the unfolded protein response", Mol Cell, 5, pp.897~904 (2000)
- Wang XZ, "Cloning of mammalian Ire1 reveals diversity in the ER stress responses", EMBO J, 17, pp.5708~5717 (1998)
- Yoshida H, "Identifi cation of the cis-acting endoplasmic reticulum stress response element responsible for transcriptional induction of mammalian glucose- regulated proteins. Involvement of basic leucine zipper transcription factors", J Biol Chem, 273, pp.33741~33749 (1998)
- Oyadomari S, "Roles of CHOP", GADD153 in endoplasmic reticulum stress, 2004, pp.11, 381~389
- Chami M, "Paterlini- Bréchot P. Role of SERCA1 Truncated Isoform in the Proapoptotic Calcium Transfer from ER to Mitochondria during ER Stress", Mol Cell, 32, pp.641~651 (2008)
- Qi X, "deltaPKC participates in the endoplasmic reticulum stress-induced response in cultured cardiac myocytes and ischemic heart", J Mol Cell Cardiol, 43, pp.420~428 (2007)
- Tajiri S, "Ischemia-induced neuronal cell death is mediated by the endoplasmic reticulum stress pathway involving CHOP", Cell Death Differ, 11, pp.403~415 (2004)
- Florea SM, "The effect of oxidative stress on Ca2+ release and capacitative Ca2+ entry in vascular endothelial cells", Cell Calcium, 43, pp.405~415 (2008)
- Xue X, "Tumor necrosis factor alpha (TNFalpha) induces the unfolded protein response (UPR) in a reactive oxygen species (ROS)-dependent fashion, and the UPR counteracts ROS accumulation by TNFalpha", J Biol Chem, 280, pp.33917~33925 (2005)
- Feghali JG, "L-n-acetyl-cysteine protection against cisplatin-induced auditory neuronal and hair cell toxicity", Laryngoscope, 111, pp.1147~1155 (2001)
- Jordan P, "Molecular mechanisms involved in cisplatin cytotoxicity", Cell Mol Life Sci, 57, pp.1229~1235 (2000)
- Kartalou M, "Mechanisms of resistance to cisplatin", Mutat Res, 478, pp.23~43 (2001)