Pseudomonas aeruginosa Exotoxin A Reduces Chemoresistance of Oral Squamous Carcinoma Cell via Inhibition of Heat Shock Proteins 70 (HSP70)
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Purpose: Oral squamous carcinoma (OSCC) cells exhibit resistance to chemotherapeutic agent-mediated apoptosis in the late stage of malignancy. Increased levels of heat shock proteins 70 (HSP70) in cancer cells are known to confer resistance to apoptosis. Since recent advances in the understanding of bacterial toxins have produced new strategies for the treatment of cancers, we investigated the effect of Pseudomonas aeruginosa exotoxin A (PEA) on HSP70 expression and induction of apoptosis in chemoresistant OSCC cell line (YD-9). Materials and Methods:The apoptotic effect of PEA on chemoresistant YD-9 cells was confirmed by MTT,Hoechst and TUNEL stains, DNA electrophoresis, and Western blot analysis. Results: While YD-9 cells showed high resistance to chemotherapeutic agents such as etoposide and 5-fluorouraci (5-FU), HSP70 antisense oligonucelotides sensitized chemoresistant YD-9 cells to etoposide and 5-FU. On the other hand, PEA significantly decreased the viability of YD-9 cells by deteriorating the HSP70-relating protecting system through inhibition of HSP70 expression and inducing apoptosis in YD-9 cells. Apoptotic manifestations were evidenced by changes in nuclear morphology, generation of DNA fragmentation, and activation of caspases. While p53, p21, and E2F-1 were upregulated, cdk2 and cyclin B were downregulated by PEA treatment, suggesting that PEA caused cell cycle arrest at the G2/M checkpoint. Conclusion: Therefore, these results indicate that PEA reduced the chemoresistance through inhibition of HSP70 expression and also induced apoptosis in chemoresistant YD-9 cells.
Abstract
Purpose: Oral squamous carcinoma (OSCC) cells exhibit resistance to chemotherapeutic agent-mediated apoptosis in the late stage of malignancy. Increased levels of heat shock proteins 70 (HSP70) in cancer cells are known to confer resistance to apoptosis. Since recent advances in the understanding of bacterial toxins have produced new strategies for the treatment of cancers, we investigated the effect of Pseudomonas aeruginosa exotoxin A (PEA) on HSP70 expression and induction of apoptosis in chemoresistant OSCC cell line (YD-9). Materials and Methods:The apoptotic effect of PEA on chemoresistant YD-9 cells was confirmed by MTT,Hoechst and TUNEL stains, DNA electrophoresis, and Western blot analysis. Results: While YD-9 cells showed high resistance to chemotherapeutic agents such as etoposide and 5-fluorouraci (5-FU), HSP70 antisense oligonucelotides sensitized chemoresistant YD-9 cells to etoposide and 5-FU. On the other hand, PEA significantly decreased the viability of YD-9 cells by deteriorating the HSP70-relating protecting system through inhibition of HSP70 expression and inducing apoptosis in YD-9 cells. Apoptotic manifestations were evidenced by changes in nuclear morphology, generation of DNA fragmentation, and activation of caspases. While p53, p21, and E2F-1 were upregulated, cdk2 and cyclin B were downregulated by PEA treatment, suggesting that PEA caused cell cycle arrest at the G2/M checkpoint. Conclusion: Therefore, these results indicate that PEA reduced the chemoresistance through inhibition of HSP70 expression and also induced apoptosis in chemoresistant YD-9 cells.
참고문헌 (45)
- D’Silva NJ, "Tissue biomarkers for diagnosis & management of oral squamous cell carcinoma", Alpha Omegan, 100, pp.182~189 (2007)
- Crowe DL, "p53 apoptotic response to DNA damage dependent on bcl2 but not bax in head and neck squamous cell carcinoma lines", Head Neck, 28, pp.15~23 (2006)
- Tong D, "5-Fluorouracil-induced apoptosis in cultured oral cancer cells", Oral Oncol, 36, pp.236~241 (2000)
- Spierings D, "Connected to death: the (unexpurgated) mitochondrial pathway of apoptosis", Science, 310, pp.66~67 (2005)
- Beere HM, "Stress management - heat shock protein-70 and the regulation of apoptosis", Trends Cell Biol, 11, pp.6~10 (2001)
- Rashmi R, "Human colon cancer cells lacking Bax resist curcumin-induced apoptosis and Bax requirement is dispensable with ectopic expression of Smac or downregulation of Bcl-XL", Carcinogenesis, 26, pp.713~723 (2005)
- Sreedhar AS, "Heat shock proteins in the regulation of apoptosis: new strategies in tumor therapy: a comprehensive review", Pharmacol Ther, 101, pp.227~257 (2004)
- Garrido C, "Heat shock proteins 27 and 70: anti-apoptotic proteins with tumorigenic properties", Cell Cycle, 5, pp.2592~2601 (2006)
- Li M, "Synergistic promotion of breast cancer cells death by targeting molecular chaperone GRP78 and heat shock protein 70", J Cell Mol Med, 13, pp.4540~4550 (2009)
- Grivicich I, "Hsp70 response to 5-fluorouracil treatment in human colon cancer cell lines", Int J Colorectal Dis, 22, pp.1201~1208 (2007)
- Bickels J, "Coley’s toxin:historical perspective", Isr Med Assoc J, 4, pp.471~472 (2002)
- Zacharski LR, "Coley’s toxin revisited: immunotherapy or plasminogen activator therapy of cancer?", J Thromb Haemost, 3, pp.424~427 (2005)
- Alley SC, "The pharmacologic basis for antibody-auristatin conjugate activity", J Pharmacol Exp Ther, 330, pp.932~938 (2009)
- Pastan I, "Immunotoxins containing Pseudomonas exotoxin A: a short history", Cancer Immunol Immunother, 52, pp.338~341 (2003)
- Koo HM, "Apoptosis and melanogenesis in human melanoma cells induced by anthrax lethal factor inactivation of mitogen-activated protein kinase kinase", Proc Natl Acad Sci U S A, 99, pp.3052~3057 (2002)
- Lee SY, "Shiga toxin 1 induces apoptosis in the human myelogenous leukemia cell line THP-1 by a caspase-8-dependent, tumor necrosis factor receptorindependent mechanism", Infect Immun, 73, pp.5115~5126 (2005)
- Cho SJ, "Induction of apoptosis and expression of apoptosis related genes in human epithelial carcinoma cells by Helicobacter pylori VacA toxin", Toxicon, 42, pp.601~611 (2003)
- Demir M, "Investigation of siderophore, total matrix protease and elastase activity in Pseudomonas aeruginosa isolates from lower respiratory tract and extra-respiratory tract samples", Mikrobiyol Bul, 42, pp.197~208 (2008)
- Marra M, "Anticancer drugs and hyperthermia enhance cytotoxicity induced by polyamine enzymatic oxidation products", Amino Acids, 33, pp.273~281 (2007)
- Kreitman RJ, "Immunotoxins for targeted cancer therapy", AAPS J, 8, pp.E532~551 (2006)
- Andersson Y, "Downregulation of the antiapoptotic MCL-1 protein and apoptosis in MA-11 breast cancer cells induced by an anti-epidermal growth factor receptor-Pseudomonas exotoxin a immunotoxin", Int J Cancer, 112, pp.475~483 (2004)
- Decker T, "Induction of caspase-dependent programmed cell death in B-cell chronic lymphocytic leukemia by anti-CD22 immunotoxins", Blood, 103, pp.2718~2726 (2004)
- 이은주, "Characterization of newly established oral cancer cell lines derived fromsix squamous cell carcinoma and two mucoepidermoid carcinoma cells", Experimental and Molecular Medicine, 37(5), pp.379~390 (2005), 생화학분자생물학회
- Zhang P, "Chemopreventive agents induce programmed death-1-ligand 1 (PD-L1) surface expression in breast cancer cells and promote PD-L1-mediated T cell apoptosis", Mol Immunol, 45, pp.1470~1476 (2008)
- Azuma M, "Potentiation of induction of apoptosis by sequential treatment with cisplatin followed by 5-fluorouracil in human oral cancer cells", Int J Oncol, 24, pp.1449~1455 (2004)
- Hsu S, "Induction of apoptosis in oral cancer cells: agents and mechanisms for potential therapy and prevention", Oral Oncol, 40, pp.461~473 (2004)
- Schümann J, "Acute hepatotoxicity of Pseudomonas aeruginosa exotoxin A in mice depends on T cells and TNF", J Immunol, 161, pp.5745~5754 (1998)
- Jenkins CE, "Pseudomonas aeruginosa exotoxin A induces human mast cell apoptosis by a caspase-8 and -3-dependent mechanism", J Biol Chem, 279, pp.37201~37207 (2004)
- Li CY, "Heat shock protein 70 inhibits apoptosis downstream of cytochrome c release and upstream of caspase-3 activation", J Biol Chem, 275, pp.25665~25671 (2000)
- Gabai VL, "Increased expression of the major heat shock protein Hsp72 in human prostate carcinoma cells is dispensable for their viability but confers resistance to a variety of anticancer agents", Oncogene, 24, pp.3328~3338 (2005)
- Meyn RE, "The role of apoptosis in radiation oncology", Int J Radiat Biol, 85, pp.107~115 (2009)
- Moretto P, "Targeting apoptosis: preclinical and early clinical experience with mapatumumab, an agonist monoclonal antibody targeting TRAIL-R1", Expert Opin Investig Drugs, 18, pp.311~325 (2009)
- 김소연, "Cell Death and Stress Signaling in Glycogen Storage Disease Type I", 분자와 세포, 28(3), pp.139~148 (2009), 한국분자세포생물학회
- Creagh EM, "Caspases: cellular demolition experts", Biochem Soc Trans, 29, pp.696~702 (2001)
- Oancea M, "Apoptosis assays", Methods Mol Med, 129, pp.279~290 (2006)
- Thant AA, "Role of caspases in 5-FU and selenium-induced growth inhibition of colorectal cancer cells", Anticancer Res, 28, pp.3579~3592 (2008)
- Wong SH, "Caspases and nitric oxide broadly regulate dendritic cell maturation and surface expression of class II MHC proteins", Proc Natl Acad Sci U S A, 101, pp.17783~17788 (2004)
- Harbour JW, "The Rb", E2F pathway: expanding roles and emerging paradigms, 2000, pp.14, 2393~2409
- Endo-Munoz L, "E2F7 can regulate proliferation, differentiation, and apoptotic responses in human keratinocytes:implications for cutaneous squamous cell carcinoma formation", Cancer Res, 69, pp.1800~1808 (2009)
- Nahle Z, "Direct coupling of the cell cycle and cell death machinery by E2F", Nat Cell Biol, 4, pp.859~864 (2002)
- Halaban R, "Deregulated E2F transcriptional activity in autonomously growing melanoma cells", J Exp Med, 191, pp.1005~1016 (2000)
- Furukawa Y, "Apaf-1 is a mediator of E2F-1-induced apoptosis", J Biol Chem, 277, pp.39760~39768 (2002)
- Stevens C, "The emerging role of E2F-1 in the DNA damage response and checkpoint control", DNA Repair (Amst), 3, pp.1071~1079 (2004)
- Li Y, "Selective killing of cancer cells by beta -lapachone: direct checkpoint activation as a strategy against cancer", Proc Natl Acad Sci U S A, 100, pp.2674~2678 (2003)
- Ismail IA, "Genisteininduced neuronal apoptosis and G2", M cell cycle arrest is associated with MDC1 up-regulation and PLK1 down-regulation, 2007, pp.575, 12~20
