Abstract Objective To study the radiosensitizing effect of 17-allylamino-17-demethoxygeldana- mycin(17-AAG) on human non-small cell lung cancer cell line (A549 cells), and to investigate its possible mechanism. Methods The growth inhibition effect of 17-AAG on A549 cells was determined using the MTT assay. The radiosensitizing effect of the drug on cells was analyzed using the multi-target single-hit model fitting the colony survival curve. The impacts of the drug on cell aging and DNA damage repair were evaluated using β-galactosidase staining and γ-H2AX immunofluorescence, respectively. Between-group comparison was performed by paired t test or analysis of variance. Results The suppression of cell proliferation by 17-AAG was both dose-dependent and time-dependent (P=0.01-0.05). The radiation group treated with the drug had a lower colony forming efficiency than the radiation-only group. When 50 nmol/L 17-AAG was applied, the radiosensitivity enhancement ratios were 1.79(the ratio of D0 values) and 2.30(the ratio of Dq values), respectively. The percentage of senescent cells at 72 hours after exposure was significantly higher in cells exposed to the drug and radiation of 4 Gy X-ray than in the drug-only group (30.48% vs. 9.18%, P=0.00) or in the radiation-only group (30.48% vs. 12.30%, P=0.00). Immunofluorescent staining showed that 17-AAG delayed the DNA damage repair. Conclusions17-AAG has growth inhibition and radiosensitizing effects on human lung cancer A549 cells. Induction of cell aging and delay of DNA damage repair may be involved in the radiosensitizing mechanism.
Wang ye*,Xue Lian,Yu Dong. Effect of 17-allylamino-17-demethoxygeldanamycin on radiosensitivity of human lung cancer cell line[J]. Chinese Journal of Radiation Oncology, 2015, 24(4): 471-473.
Wang ye*,Xue Lian,Yu Dong. Effect of 17-allylamino-17-demethoxygeldanamycin on radiosensitivity of human lung cancer cell line[J]. Chinese Journal of Radiation Oncology, 2015, 24(4): 471-473.
[1] Belani CP,Ramanathan RK.Combined-modality treatment of locally advanced non-small cell lung cancer-Incorporation of novel chemotherapeutic agents[J].Chest,1998,113(1):53-60. [2] 蒋国梁,仇德惠,谭黎杰.肺癌[A]//汤钊猷.现代肿瘤学[M].2版.上海:上海医科大学出版社,2000:859-895. [3] Kamal A,Boehm MF,Burrows FJ.Therapeutic and diagnostic implications of Hsp90 activation[J].Trends Mol Med,2004,10(6):283-290. [4] Whitesell L,Lindquist SL.HSP90 and the chaperoning of cancer[J].Nat Rev Cancer,2005,5(10):761-772. [5] Bisht KS,Bradbury CM,Mattson D,et al. Geldanamycin and 17-allylamino-17-demethoxygeldanamycin potentiate the in vitro and in vivo radiation response of cervical tumor cells via the heat shock protein 90-mediated intracellular signaling and cytotoxicity[J].Cancer Res,2003,63(24):8984-8995. [6] Enmolon R,Yang WH,Ballangrud AM,et al. Combination treatment with 17-N-allylamino-17-demethoxy geldanamycin and acute irradiation produces supra-additive growth suppression in human prostate carcinoma spheroids[J].Cancer Res,2003,63(23):8393-8399. [7] Wright L,Barril X,Dymock B,et al. Structure-activity relationships in purine-based inhibitor binding to HSP90 isoforms[J].Chem Biol,2004,11(6):775-785. [8] Harashima K,Akimoto T,Nonaka T,et al. Heat shock protein 90(Hsp90) chaperone complex inhibitor,radicicol,potentiated radiation-induced cell killing in a hormone-sensitive prostate cancer cell line through degradation of the androgen receptor[J].Int J Radiat Biol,2005,81(1):63-76. [9] Noguchi M,Yu D,Hirayama R,et al. Inhibition of homologous recombination repair in irradiated tumor cells pretreated with Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin[J].Biochem Biophys Res Commun,2006,351(3):658-663. [10] Russell JS,Brady K,Burgan WE,et al. Gleevec-mediated inhibition of Rad51 expression and enhancement of tumor cell radiosensitivity[J].Cancer Res,2003,63(21):7377-7383. [11] Musha A,Yoshida Y,Takahashi T,et al. Synergistic effect of heat shock protein 90 inhibitor,17-allylamino-17-demethoxygeldanamycin and X-rays,but not carbon-ion beams,on lethality in human oral squamous cell carcinoma cells[J].J Radiat Res,2012,53(4):545-550.DOI:10.1093/jrr/rrs012. [12] Takahashi T,Yoshida Y,Ando K,et al. Signal transduction and heavy ion radiation therapy:biological mechanisms,biological quality assurance,and new multimodality approach[J].Current Signal Transduction Therapy,2010,5(3):237-243.DOI:10.2174/157436210791920238. [13] Wang HY,Zhang XM,Wang P,et al. Characteristics of DNA-binding proteins determine the biological sensitivity to high-linear energy transfer radiation[J].Nucleic Acids Res,2010,38(10):3245-3251.DOI:10.1093/nar/gkq069. [14] Cohen-Jonathan E,Bernhard EJ,Mckenna WG.How does radiation kill cells?[J].Curr Opin Chem Biol,1999,3(1):77-83. [15] Sancar A,Lindsey-Boltz LA,Unsal-Kacmaz K,et al. Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints[J].Annu Rev Biochem,2004,73:39-85. [16] Rogakou EP,Pilch DR,Orr AH,et al. DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139[J].J Biol Chem,1998,273(10):5858-5868. [17] Rothkamm K,Lobrich M.Evidence for a lack of DNA double-strand break repair in human cells exposed to very low x-ray doses[J].Proc Natl Acad Sci USA,2003,100(9):5057-5062. [18] Rogakou EP,Boon C,Redon C,et al. Megabase chromatin domains involved in DNA double-strand breaks in vivo[J].J Cell Biol,1999,146(5):905-915. [19] Macphail SH,Banath JP,Yu TY,et al. Expression of phosphorylated histone H2AX in cultured cell lines following exposure to X-rays[J].Int J Radiat Biol,2003,79(5):351-358. [20] Fernandez-Capetillo O,Lee A,Nussenzweig M,et al. H2AX:the histone guardian of the genome[J].DNA Repair,2004,3(8-9):959-967. [21] Stecklein SR,Kumaraswamy E,Behbod F,et al. BRCA1 and HSP90 cooperate in homologous and non-homologous DNA double-strand-break repair and G2/M checkpoint activation[J].Proc Natl Acad Sci USA,2012,109(34):13650-13655.DOI:10.1073/pnas.1203326109. [22] Barker CR,Hamlett J,Pennington SR,et al. The topoisomerase Ⅱ-Hsp90 complex:a new chemotherapeutic target?[J].Int J Cancer,2006,118(11):2685-2693. [23] Faried LS,Faried A,Kanuma T,et al. Inhibition of the mammalian target of rapamycin (mTOR) by rapamycin increases chemosensitivity of CaSki cells to paclitaxel[J].Eur J Cancer,2006,42(7):934-947. [24] Weng SH,Tseng SC,Huang YC,et al. Inhibition of thymidine phosphorylase expression by using an HSP90 inhibitor potentiates the cytotoxic effect of cisplatin in non-small-cell lung cancer cells[J].Biochem Pharmacol,2012,84(1):126-136.DOI:10.1016/j.bcp.2012.03.011. [25] Neckers L.Hsp90 inhibitors as novel cancer chemotherapeutic agents[J].Trends Mol Med,2002,8(4):55-61. [26] Powers MV,Workman P.Targeting of multiple signalling pathways by heat shock protein 90 molecular chaperone inhibitors[J].Endocr Relat Cancer,2006,13 Suppl 1:S125-S135. [27] Hostein I,Robertson D,Distefano F,et al. Inhibition of signal transduction by the Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin results in cytostasis and apoptosis[J].Cancer Res,2001,61(10):4003-4009. [28] Powers MV,Clarke PA,Workman P.Dual targeting of HSC70 and HSP72 inhibits HSP90 function and induces tumor-specific apoptosis[J].Cancer Cell,2008,14(3):250-262.DOI:10.1016/j.ccr.2008.08.002. [29] Powers MV,Valenti M,Miranda S,et al. Mode of cell death induced by the HSP90 inhibitor 17-AAG (tanespimycin) is dependent on the expression of pro-apoptotic BAX[J].Oncotarget,2013,4(11):1963-1975. [30] Hasenstein JR,Shin HC,Kasmerchak K,et al. Antitumor activity of Triolimus:a novel multidrug-loaded micelle containing paclitaxel,rapamycin,and 17-AAG[J].Mol Cancer Ther,2012,11(10):2233-2242.DOI:10.1158/1535-7163. [31] Larson N,Greish K,Bauer H,et al. Synthesis and evaluation of poly (styrene-co-maleic acid) micellar nanocarriers for the delivery of tanespimycin[J].Int J Pharm,2011,420(1):111-117.DOI:10.1016/j.ijpharm.2011.08.011. [32] Saxena V,Hussain MD.Formulation and in vitro evaluation of 17-allyamino-17-demethoxygeldanamycin (17-AAG) loaded polymeric mixed micelles for glioblastoma multiforme[J].Colloids Surf B Biointerfaces,2013,112:350-355.DOI:10.1016/j.colsurfb.2013.07.031.
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