Abstract
Purpose
Flavopiridol is known to modulate the transcription of genes. We investigated the effect of flavopiridol pretreatment on TRAIL cytotoxicity and on the expression of FLIPL in different TRAIL-resistant cell lines, because FLIP expression is known to confer TRAIL-resistance.
Methods
Apoptosis was assessed by PI staining and protein expression by Western blotting. RT-PCR was used for mRNA quantitation. siRNA gene silencing was used to knock down FLIPL.
Results
Flavopiridol pretreatment synergized TRAIL-induced apoptosis in human myeloma and breast cancer cells. Flavopiridol treatment repressed the transcription of FLIPL and downregulated its expression in both myeloma and breast cancer cells. Silencing of FLIPL gene by siRNA sensitized myeloma cells to TRAIL. Flavopiridol treatment downregulated the expression of the proapoptotic members of the Bcl-2 family proteins (Bak, Bax and PUMA-α). The expression of the antiapototic Bcl-2 members (Bcl-2 and Bcl-XL) was not altered by flavopiridol treatment in myeloma cells.
Conclusion
Our data indicate that flavopiridol synergizes TRAIL cytotoxicity by downregulation of FLIPL and this synergistic effect is Bcl-2 family independent.
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References
Bible KC, Kaufmann SH (1996) Flavopiridol: a cytotoxic flavone that induces cell death in noncycling A549 human lung carcinoma cells. Cancer Res 56:4856–4861
Drees M, Dengler WA, Roth T, Labonte H, Mayo J, Malspeis L, Grever M, Sausville EA, Fiebig HH (1997) Flavopiridol (L86–8275): selective antitumor activity in vitro and activity in vivo for prostate carcinoma cells. Clin Cancer Res 3:273–279
Carlson BA, Dubay MM, Sausville EA, Brizuela L, Worland PJ (1996) Flavopiridol induces G1 arrest with inhibition of cyclin-dependent kinase (CDK) 2 and CDK4 in human breast carcinoma cells. Cancer Res 56:2973–2978
Shapiro GI, Supko JG, Patterson A, Lynch C, Lucca J, Zacarola PF, Muzikansky A, Wright JJ, Lynch TJ Jr, Rollins BJ (2001) A phase II trial of the cyclin-dependent kinase inhibitor flavopiridol in patients with previously untreated stage IV non-small cell lung cancer. Clin Cancer Res 7:1590–1599
Schwartz GK, Ilson D, Saltz L, O’Reilly E, Tong W, Maslak P, Werner J, Perkins P, Stoltz M, Kelsen D (2001) Phase II study of the cyclin-dependent kinase inhibitor flavopiridol administered to patients with advanced gastric carcinoma. J Clin Oncol 19:1985–1992
Chao SH, Fujinaga K, Marion JE, Taube R, Sausville EA, Senderowicz AM, Peterlin BM, Price DH (2000) Flavopiridol inhibits P-TEFb and blocks HIV-1 replication. J Biol Chem 275:28345–28348
Melillo G, Sausville EA, Cloud K, Lahusen T, Varesio L, Senderowicz AM (1999) Flavopiridol, a protein kinase inhibitor, down-regulates hypoxic induction of vascular endothelial growth factor expression in human monocytes. Cancer Res 59:5433–5437
Schwartz GK, Farsi K, Maslak P, Kelsen DP, Spriggs D (1997) Potentiation of apoptosis by flavopiridol in mitomycin-C-treated gastric and breast cancer cells. Clin Cancer Res 3:1467–1472
Kim DM, Koo SY, Jeon K, Kim MH, Lee J, Hong CY, Jeong S (2003) Rapid induction of apoptosis by combination of flavopiridol and tumor necrosis factor (TNF)-alpha or TNF-related apoptosis-inducing ligand in human cancer cell lines. Cancer Res 63:621–626
Rosato RR, Dai Y, Almenara JA, Maggio SC, Grant S (2004) Potent antileukemic interactions between flavopiridol and TRAIL/Apo2L involve flavopiridol-mediated XIAP downregulation. Leukemia 18:1780–1788
Irmler M, Thome M, Hahne M, Schneider P, Hofmann K, Steiner V, Bodmer JL, Schroter M, Burns K, Mattmann C et al (1997) Inhibition of death receptor signals by cellular FLIP. Nature 388:190–195
Shain KH, Landowski TH, Dalton WS (2002) Adhesion-mediated intracellular redistribution of c-Fas-associated death domain-like IL-1-converting enzyme-like inhibitory protein-long confers resistance to CD95-induced apoptosis in hematopoietic cancer cell lines. J Immunol 168:2544–2553
Srivastava RK (2001) TRAIL/Apo-2L. Mechanisms and clinical applications in cancer. Neoplasia 3:535–546
Zhang X, Jin TG, Yang H, DeWolf WC, Khosravi-Far R, Olumi AF (2004) Persistent c-FLIP(L) expression is necessary and sufficient to maintain resistance to tumor necrosis factor-related apoptosis-inducing ligand-mediated apoptosis in prostate cancer. Cancer Res 64:7086–7091
Zeise E, Weichenthal M, Schwarz T, Kulms D (2004) Resistance of human melanoma cells against the death ligand TRAIL is reversed by ultraviolet-B radiation via downregulation of FLIP. J Invest Dermatol 123:746–754
Chang DW, Xing Z, Pan Y, Algeciras-Schimnich A, Barnhart BC, Yaish-Ohad S, Peter ME, Yang X (2002) c-FLIP(L) is a dual function regulator for caspase-8 activation and CD95-mediated apoptosis. EMBO J 21:3704–3714
Micheau O, Thome M, Schneider P, Holler N, Tschopp J, Nicholson DW, Briand C, Grutter MG (2002) The long form of FLIP is an activator of caspase-8 at the Fas death-inducing signaling complex. J Biol Chem 277:45162–45171
Fandy TE, Shankar S, Ross DD, Sausville E, Srivastava RK (2005) Interactive effects of HDAC inhibitors and TRAIL on apoptosis are associated with changes in mitochondrial functions and expressions of cell cycle regulatory genes in multiple myeloma. Neoplasia 7:646–657
Singh TR, Shankar S, Chen X, Asim M, Srivastava RK (2003) Synergistic interactions of chemotherapeutic drugs and tumor necrosis factor-related apoptosis-inducing ligand/Apo-2 ligand on apoptosis and on regression of breast carcinoma in vivo. Cancer Res 63:5390–5400
Srivastava RK, Mi QS, Hardwick JM, Longo DL (1999) Deletion of the loop region of Bcl-2 completely blocks paclitaxel-induced apoptosis. Proc Natl Acad Sci USA 96:3775–3780
Srivastava RK, Sollott SJ, Khan L, Hansford R, Lakatta EG, Longo DL (1999) Bcl-2 and Bcl-X(L) block thapsigargin-induced nitric oxide generation, c-Jun NH(2)-terminal kinase activity, and apoptosis. Mol Cell Biol 19:5659–5674
Burns TF, El-Deiry WS (2001) Identification of inhibitors of TRAIL-induced death (ITIDs) in the TRAIL-sensitive colon carcinoma cell line SW480 using a genetic approach. J Biol Chem 276:37879–37886
Narita M, Shimizu S, Ito T, Chittenden T, Lutz RJ, Matsuda H, Tsujimoto Y (1998) Bax interacts with the permeability transition pore to induce permeability transition and cytochrome c release in isolated mitochondria. Proc Natl Acad Sci USA 95:14681–14686
Konig A, Schwartz GK, Mohammad RM, Al-Katib A, Gabrilove JL (1997) The novel cyclin-dependent kinase inhibitor flavopiridol downregulates Bcl-2 and induces growth arrest and apoptosis in chronic B-cell leukemia lines. Blood 90:4307–4312
Robinson WA, Miller TL, Harrold EA, Bemis LT, Brady BM, Nelson RP (2003) The effect of flavopiridol on the growth of p16+ and p16− melanoma cell lines. Melanoma Res 13:231–238
Schrump DS, Matthews W, Chen GA, Mixon A, Altorki NK (1998) Flavopiridol mediates cell cycle arrest and apoptosis in esophageal cancer cells. Clin Cancer Res 4:2885–2890
Byrd JC, Shinn C, Waselenko JK, Fuchs EJ, Lehman TA, Nguyen PL, Flinn IW, Diehl LF, Sausville E, Grever MR (1998) Flavopiridol induces apoptosis in chronic lymphocytic leukemia cells via activation of caspase-3 without evidence of bcl-2 modulation or dependence on functional p53. Blood 92:3804–3816
Achenbach TV, Muller R, Slater EP (2000) Bcl-2 independence of flavopiridol-induced apoptosis. Mitochondrial depolarization in the absence of cytochrome c release. J Biol Chem 275:32089–32097
Palacios C, Yerbes R, Lopez-Rivas A (2006) Flavopiridol induces cellular FLICE-inhibitory protein degradation by the proteasome and promotes TRAIL-induced early signaling and apoptosis in breast tumor cells. Cancer Res 66:8858–8869
Gojo I, Zhang B, Fenton RG (2002) The cyclin-dependent kinase inhibitor flavopiridol induces apoptosis in multiple myeloma cells through transcriptional repression and down-regulation of Mcl-1. Clin Cancer Res 8:3527–3538
Taniai M, Grambihler A, Higuchi H, Werneburg N, Bronk SF, Farrugia DJ, Kaufmann SH, Gores GJ (2004) Mcl-1 mediates tumor necrosis factor-related apoptosis-inducing ligand resistance in human cholangiocarcinoma cells. Cancer Res 64:3517–3524
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Fandy, T.E., Ross, D.D., Gore, S.D. et al. Flavopiridol synergizes TRAIL cytotoxicity by downregulation of FLIPL . Cancer Chemother Pharmacol 60, 313–319 (2007). https://doi.org/10.1007/s00280-006-0381-8
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DOI: https://doi.org/10.1007/s00280-006-0381-8