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Histone Deacetylase Inhibitor Potentiates Anticancer Effect of Docetaxel via Modulation of Bcl-2 Family Proteins and Tubulin in Hormone

Refractory Prostate Cancer Cells
Jung Jin Hwang, Yong Sook Kim, Mi Joung Kim, Dong Eun Kim, In Gab Jeong and Choung-Soo Kim*
From the Institute for Innovative Cancer Research (JJH, YSK, MJK, DEK) and Department of Urology (IGJ), University of Ulsan College of Medicine, Asan Medical Center (CSK), Seoul, Korea

Purpose: We evaluated the antitumor effects of docetaxel (Sigma®) and histone deacetylase inhibitors in hormone refractory prostate cancer cells, and analyzed the mechanism by which combination treatment induced cell death. Materials and Methods: We used LNCaP, DU145 and PC3 cells (ATCC®) to evaluate the in vitro apoptotic effects of histone deacetylase inhibitors and their combinations with docetaxel as well as the molecular mechanisms. The DU145 xenograft model was used to evaluate the in vivo efficacy of PXD101 combined with docetaxel. Results: Suberoylanilide hydroxamic acid or PXD101 inhibited the growth of hormone dependent LNCaP cells, and hormone independent DU145 and PC3 cells. It increased sub-G1 population and activated caspase-8, 9 and 3, indicating apoptosis induction. Pretreating DU145 cells with docetaxel followed by histone deacetylase inhibitors showed significant synergistic cytotoxicity compared with that of simultaneous co-treatment or reverse sequential treatment. Pretreatment with docetaxel followed by histone deacetylase inhibitors increased the apoptotic sub-G1 population, caspase activation and tubulin acetylation compared with that of docetaxel alone. Combination treatment decreased Mcl-1 and Bcl-xl, and increased t-Bid, Bik and Bim. Combined docetaxel and PXD101 reduced tumor size with efficacy equivalent to that of a double dose of docetaxel alone in the DU145 xenograft model. Conclusions: These preclinical results indicate that the sequential combination of docetaxel and histone deacetylase inhibitors led to a synergistic increase in the death of hormone refractory prostate cancer cells via intrinsic and extrinsic apoptotic pathways by modulating Bcl-2 family proteins and tubulin in vitro and in vivo. Results suggest that this combination may be a new therapeutic modality in patients with hormone refractory prostate cancer. Key Words: prostate, prostatic neoplasms, histone deacetylases, docetaxel, apoptosis PROSTATE cancer is the most common nonskin cancer and the second most common cause of cancer death in men in the United States with an estimated 186,320 newly diagnosed patients and 27,360 prostate cancer deaths in 2008.1
0022-5347/10/1846-2557/0 THE JOURNAL OF UROLOGY® © 2010 by AMERICAN UROLOGICAL ASSOCIATION EDUCATION

Abbreviations and Acronyms CI combination index histone deacetylase HDAC inhibitor HDAC HDACI

HRPC hormone refractory prostate cancer MTS 3-(4,5-dimethylthiazol-2yl)-5-(3-carboxymethoxyphenyl)-2(4-sulfophenyl)-2H-tetrazolium SAHA suberoylanilide hydroxamic acid v/v volume per volume

Submitted for publication March 23, 2010. Study received institutional animal care and use committee approval. Supported by Grant A062254 from the Korea Health 21 R & D Project, Ministry of Health, Welfare and Family Affairs, and a grant (2009-450) from the Asan Institute for Life Sciences, Republic of Korea. * Correspondence: Department of Urology, Asan Medical Center, 388-1 Pungnap 2 dong, Songpa-gu, Seoul 138-736, Korea (telephone: 82-23010-3734; FAX: 82-2-477-8928; e-mail: cskim@ amc.seoul.kr).

Although docetaxel based regimens have palliative and survival benefits, men with metastatic HRPC have only 16 to 18-month median survival.2,3 Thus, new therapeutic modalities aimed to improve manVol. 184, 2557-2564, December 2010 Printed in U.S.A. DOI:10.1016/j.juro.2010.07.035

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including the hydroxamic acids SAHA. A numerical CI value is calculated based on the equation. We measured the percent of 10.20 –22 However. The isobologram is a graphic representation of the interaction between 2 drugs that is formed by plotting the individual drug doses required to achieve a single agent effect on the respective x and y-axes. Of these modifications the acetylation of lysine residues at the histone N-terminal tail decreases the affinity of histone for DNA.16 –19 the precise mechanism of HDACIs must be further elucidated. Similar to the isobologram. death ligands and retinoic acid receptor. Cell Cycle Analysis Cells were treated with drugs. SAHA or PXD101 for solid tumors. ubiquitination or adenosine triphosphate ribosylation. PXD101 and FR235222 decreased prostate specific antigen. Combination data points that fall on the line represent an additive interaction while points above and below represent antagonism and synergy. CI (D)1/(Dx)1 (D)2/(Dx)2 (D)1(D)2/(Dx)1(Dx)2. DU145 and PC3 in RPMI 1640 medium containing 100 U/ml penicillin. breast and lung cancer. phosphorylation.13–15 Although it was reported that SAHA. HDACIs show potent activity against many cancers. apoptotic Bcl-2 family proteins. with the same certain percent effect when used alone. Immobilon™ Western ECL solution and Image Station 4000MM . respectively.7–10 Many reports show the usefulness of various HDACIs for prostate cancer. PXD101 or docetaxel in 5% (v/v) RPMI 1640 medium containing fetal bovine serum. LBH 589 and trichostatin A. Appropriate secondary antibodies (1:5. After blocking with 3% (weight per volume) nonfat dry milk the membranes were incubated with various primary antibodies (1:1. including tubulin. methylation. which is based on the Chou and Talalay median effect principle. A line connecting the 2 points is drawn and the concentration of the 2 drugs used in combination to achieve the same effect is plotted on the isobologram. fixed with 70% (v/v) ethanol and stained with 60 g/ml propidium iodide (Sigma) containing 10 U/ml ribonuclease A for 30 minutes. PXD101. resulting in the expression of genes related to tumor suppression and/or differentiation. apicidin and depsipeptide (FK 228). Illinois) were incubated for 1 hour at room temperature. We measured viability with the CellTiter 96® Aqueous One Solution Cell proliferation assay (MTS assay).5 Thus. respectively. Finally.2558 HISTONE DEACETYLASE INHIBITOR POTENTIATES DOCETAXEL IN PROSTATE CANCER agement for advanced HRPC include docetaxel plus other agents. 1 indicates an additive effect and less than 1 indicates synergy. disrupting protein function and resulting in cell death. 100 g/ml streptomycin and 10% (v/v) fetal bovine serum (Invitrogen™).9. death receptors. CI analysis provides qualitative information on the drug interaction.11. the shortchain fatty acids 4-phenylbutyrate.12 HDACI also acetylates many nonhistone proteins. We treated cells with SAHA. MS-275. or that methylate DNA. The HDACIs SAHA and PXD101 were synthesized elsewhere. and benzamide (MS-275).000) conjugated to horseradish peroxidase (Pierce. Rockford. including prostate. for example the taxane paclitaxel or docetaxel combined with the HDACI pivaloyloxymethyl butyrate. we determined the antitumor effect of docetaxel and HDACI in HRPC DU145 cells in vitro and in vivo. Western Blot Equal amounts of protein were electrophoresed in sodium dodecyl sulfate-polyacrylamide gel and transferred to Millipore polyvinylidene difluoride membranes (Chemicon®). where (D)1 and (D)2 are the doses of drugs 1 and 2 with a certain percent effect when used in combination. and analyzed the mechanism by which combination treatment induced cell death. Thus. LBH589. the cyclic tetrapeptides trapoxin. and (Dx)1 and (Dx)2 are the doses of drugs 1 and 2. heat shock proteins and Ku70.7 Many clinical trials have been done or are under way using HDACIs combined with other chemotherapy.000) in blocking solution overnight at 4C. we evaluated the growth inhibitory effects of HDACI alone in hormone dependent and independent prostate cancer cells. and p21 and annexin A1 expression in a prostate cancer cell line. the exact mechanism of action is not fully understood.000 cells in the different cell cycle phases using the FACSCalibur™ flow cytometer built-in ModFit LT™ 3. We also assessed the mechanism of cell death. For sequential combination treatment with HDACI and docetaxel cells were exposed to the former drug for 24 hours and then to the next drug for another 48 hours. We determined whether HDACIs would enhance the effects of docetaxel in advanced prostate cancer cells.6 Five HDACI classes have been characterized. MTS reagent was added to each well according to manufacturer instructions.8. HDACIs are well tolerated with a good toxicity profile compared to that of other anticancer agents. pivaloyloxymethyl butyrate and valproic acid.11 Cell death mechanisms of HDACIs include induction of the cyclin dependent kinase inhibitors p21 and p27. MATERIALS AND METHODS Cell Culture and Drug Treatment We cultured the human prostate cancer cell lines LNCaP. and increased caspase activation. valproic acid. LAQ-824.0 software. Cell Survival and CI Analysis Cells were cultured in 96-well plates and treated with various drug concentrations for the indicated times. We calculated the CI with CalcuSyn (Biosoft®).4 The importance of epigenetic changes in cancer cells led to the development of agents that modify histones via acetylation. After 2-hour incubation we determined cell viability by measuring absorbance at 490 nm. The merit of HDACIs is that cytotoxic effects are specific to cancer cells and not to normal cells or tissues. CI greater than 1 indicates antagonism.

HISTONE DEACETYLASE INHIBITOR POTENTIATES DOCETAXEL IN PROSTATE CANCER 2559 (Kodak. 5. B. HDACIs increased acetylated tubulin (Ac-tubulin) in cells on 15% gel.80 and 0. Conc.2. C. tubulin.20 M in PC3 cells. percent viability of 4 preparations of each cell type exposed to 0. The half maximum inhibitory concentration of SAHA and PXD101 was 2. Tumor volume was measured twice weekly and calculated using the formula.m. respectively. concentration.60 and 1. p21 (Cell Signaling Technology®). 2. PXD101 was more potent than SAHA in all preparations.05 and determined by 1-way ANOVA. respectively. DU145 and PC3 cell lines. histone 3 acetylation (Ac-H3) was increased by HDACIs in cells exposed to 1 or 10 M SAHA or PXD101. These results indicate that hormone dependent and independent prostate cancer cells are sensitive to HDACIs.) for 3 weeks. 5. 0. To assess the effects of these 2 HDACIs on intracellular HDAC activity we analyzed histone 3 and tubulin acetylation by Western blot in the 3 cell lines Xenograft Animal Model Four-week-old male BALB/C nude mice (OrientBio. Mice bearing tumors with a volume of about 100 mm3 intraperitoneally received docetaxel (10 or 5 mg/kg for 1 day per week at 9:00 a. Korea) were subcutaneously inoculated with 5 106 DU145 cells. 1.70 M in DU145 cells. Figure 1. tumor volume in mm3 1/2 ( 1 x 22). Santa Cruz. actin. Mcl-1 and Bcl-xl (Santa Cruz Biotechnology. 25 or 100 M SAHA or PXD101 for 48 hours decreased cell viability in a dose dependent manner (fig. 1.75 M in LNCaP cells. 1. Statistical significance was considered at p 0. Bid. Rochester. Statistical Analysis All data are shown as the mean SD.04. histone 3. Cytotoxicity and activity of HDACIs SAHA and PXD101 in LNCaP.m. caspase-3. . 0. acetylated tubulin (Sigma). Seoul. and DU145 and PC3 HRPC cells with 0. A). California). 1. New York) were used to visualize immunoreactive bands. h. Bik.2. 8 and 9. Incubating hormone dependent LNCaP prostate cancer cells. A). hours. where 1 and 2 represent the larger and smaller tumor diameters. Bim. RESULTS HDAC Inhibitors Prostate cancer cell growth inhibition. We used antibodies to acetylated histone 3. 25 and 100 M SAHA or PXD101 for 48 hours. and 6.04. DU145 cells were as sensitive as LNCaP and more sensitive than PC3 cells (fig.) and PXD101 (30 mg/kg for 5 days per week at 6:00 p.50 and 0. A.

A). J and K). and the activation of caspase-9 and 3. DU145 and PC3 cells treated with 10 M SAHA or PXD101 for 48 hours (fig. Cell growth was mark- edly inhibited by docetaxel and HDACI applied in combination compared with that of each drug alone. flow cytometry profiles show that HDACI increased sub-G1 apoptotic population of cells treated with 10 M SAHA or PXD101 for 48 hours. This indicates that combined docetaxel and HDACI treatment (docetaxel¡HDACIs) increased apoptosis by modulating Bcl-2 family protein expression. 65. H and I). 9 and 3 in cells treated with 5 M SAHA or PXD101 for 72 hours on 15% gel. 4. PXD101. B. 3. which mediate intrinsic apoptosis. E and F).6% 7. B and C).5 M SAHA or 0. We observed an increase in the hypodiploid population (sub-G1) of LNCaP.8% 8.05 vs control. The histone 3 or tubulin level was not changed by HDACIs. respectively. DU145 and PC3 cells. Histograms showed that combination treatment increased the sub-G1 population compared with cells exposed to a single agent.5 M SAHA or 0. which are sensitive to HDACIs. CTL. 3. A. SAHA and PXD101 increased histone 3 and tubulin acetylation in a time and dose dependent manner. Induction of DU145 cell apoptosis with docetaxel. To investigate apoptosis induced by combination treatment we performed flow cytometry of DU145 cells sequentially exposed to docetaxel and HDACIs. To establish whether the combined effects of docetaxel. respectively). 3. B and C). Effect on cell cycle and HRPC cell apoptotic signaling. adding docetaxel had no effect on the level of acetylation on histone 3 due to HDACIs. 3. Bik and Bim). we tested the effect of HDACIs on docetaxel toxicity in DU145 cells. Since combination treatment also significantly increased activated caspase-8. 3.5 M PXD101 alone resulted in modest toxicity (mean SD 69. Since HDACI had an antiproliferative effect on HRPC cells. .6% viability.8 Figure 2. 3. A). we analyzed the level of proteins that regulate cell survival and death (fig.7% viability. However.0% and 35. Although 1. On the other hand.5 M PXD101 slightly increased tubulin acetylation. Growth inhibition was then measured by MTS assay (fig. fig. The docetaxel and PXD101 combination (docetaxel ¡PXD101) similarly showed synergy except for the highest concentration of the combination on isobologram (fig. 1. Exposure of these cells to 5 M SAHA or PXD101 for 72 hours resulted in activation of caspase-8. Combination Treatment Apoptosis inducing mechanisms.5%. 1.2560 HISTONE DEACETYLASE INHIBITOR POTENTIATES DOCETAXEL IN PROSTATE CANCER exposed to 1 or 10 M SAHA or PXD101 for the indicated times (fig. C). sub-G1 population of 3 cell lines in 3 preparations each. Asterisk indicates p 0. and the HDACIs SAHA and PXD101 were synergistic we exposed DU145 cells to the drugs while keeping a constant ratio of each drug to the other.5 nM docetaxel. control. PXD. Western blots of caspase-8. 4. Since pan-HDACIs and docetaxel increase in tubulin acetylation. administering HDACIs followed by docetaxel (HDACIs ¡docetaxel) had no effect on cell death while simultaneous treatment with HDACIs and docetaxel was less effective than docetaxel followed by HDACIs (fig. C. as reported previously. SAHA or PXD101 (fig. 2. Analysis of the dose effect relationship and isobolograms of these results revealed that sequential treatment with a high concentration of docetaxel and SAHA (docetaxel ¡SAHA) was synergistic (fig. 9 and 3. 1. combined docetaxel/HDACIs increased it strongly. To assess the effects of these HDACIs on the cell cycle we analyzed DNA contents by flow cytometry.0% 3. that is docetaxel.3% 3. pretreatment with 1.5 M SAHA or 0. A and B). D and G). Apoptosis induction by SAHA or PXD101 in LNCaP.5 nM docetaxel followed by 1.5 nM docetaxel. an enzyme with a role in the extrinsic apoptotic pathway (fig. 2. The cell cycle inhibitor p21 protein was accumulated by combination treatment. Although treatment with 1. we examined the accumulation of tubulin acetylation in DU145 cells using combination treatment (fig. B and C).7% and 73% 2. Combination treatment decreased levels of the anti-apoptotic Bcl-2 family proteins (Mcl-1 and Bcl-xl) and increased levels of pro-apoptotic Bcl-2 family proteins (t-Bid.5 M PXD101 increased toxicity significantly (35.

6 preparations each of cells were incubated with 1. A. Cytotoxicity of combined HDACI and docetaxel treatment in DU145 HRPC cells. E. .5 nM docetaxel (DTX) for 24 hours and then with or without 1.5 nM docetaxel followed by 1.5 M SAHA or 0. K.000 ratio. J. flow cytometry profiles reveal that sub-G1 apoptotic population was greater after sequential treatment with 1. B. H. viability curves in 3 cell preparations each of single agent docetaxel and SAHA.5 M PXD101 for 24 hours and then with or without 1.5 M PXD101. G. isobologram shows combined docetaxel and PXD101 (G).5 nM docetaxel simultaneously for 48 hours. isobologram shows combined docetaxel and SAHA (D). bars represent sub-G1 population of sequentially (DTX ¡ HDACI) treated cells in 3 preparations each. and sequential combinations in 3 preparations each of cells treated with docetaxel and PXD101 at 1:333 ratio. and 1. 6 preparations each of cells were exposed to HDACI.5 nM docetaxel for 48 hours (HDACI ¡ DTX). and sequential combinations in cells treated with docetaxel and SAHA at 1:1. or 0.5 M PXD101. viability curves of single agent docetaxel and PXD101.5 M SAHA or 0. HDACI posttreatment significantly potentiated docetaxel induced cell death. CI plot of the combined docetaxel and PXD101 (G).HISTONE DEACETYLASE INHIBITOR POTENTIATES DOCETAXEL IN PROSTATE CANCER 2561 Figure 3. 6 preparations each of cells were incubated with 1. I. F.5 M SAHA.5 M SAHA or 0. C. D. CTL. CI plot of combined docetaxel and SAHA (D). control.5 M PXD101 for 48 hours (DTX ¡ HDACI). that is 1.

led to the development of HDACIs as antitumor agents. Effect on DU145 xenografts. tumor volume percent reduction after treatment with 5 or 10 mg/kg per week docetaxel combined with 30 mg/kg per day PXD101 for 3 weeks. Asterisk indicates p 0. PXD101 may allow a dose reduction of docetaxel. Plus signs indicate positive. Western blot shows effect of sequential combination treatment with docetaxel followed by HDACI on histone acetylation. total body weight in 3 treatment groups (A).5 nM docetaxel for 24 hour and then next with or without 1.5 M SAHA or 0. These results suggest that when combined. Asterisk indicates p 0. Plus signs indicate positive. C).05 vs control. PXD101 alone and docetaxel alone. Bcl-2 family proteins from cells treated with sequential combination on 13% gel. caspase activation and Bcl-2 family protein expression in DU145 cells. B. A. 9 and 3 from cells treated with sequential combinations on 15% gel. DISCUSSION The finding that HDAC activity is increased in tumors. C. C. acetylated histone 3 (Ac-H3) and acetylated tubulin (Ac-Tubulin) from cells incubated with 1. A.23 Many HDACIs are in phase I/II clinical trials of treatment for hematopoietic cancer and solid tumors6 but to date only SAHA has been approved by the Food and Drug Administration in the United States as treatment for cutaneous T-cell lymphoma only. To determine whether PXD101 decreased the required dose of docetaxel we treated tumor bearing mice with 5 mg/kg per week docetaxel and/or 30 mg/kg per day PXD101 for 3 weeks and then calculated tumor size (fig. 5. Injecting 5 mg/kg per week docetaxel plus 30 mg/kg per day PXD101 re- sulted in a mean tumor volume of 64% 0. B. A and B). thus decreasing its side effects. We eval- Figure 5. maintained them until the tumor size reached 100 mm3 and then injected them intraperitoneally with docetaxel (10 mg/kg per week) and/or PXD101 (30 mg/kg per day) for 3 weeks. Combined treatment reduced tumor volume compared with that in control mice or mice treated with docetaxel or PXD101 alone but combined treatment had no effect on body weight (fig. including prostate cancer. similar to the 64% 0. To confirm the effects of docetaxel and HDACI on HRPC cells in vivo we subcutaneously injected nude mice with 5 106 DU145 cells. Minuses indicate negative.2% that in saline injected controls. Minuses indicate negative. 5.5 M PXD101 for 48 hours on 15% gel. Growth inhibitory effect of combined HDACI and docetaxel (DTX) in DU145 xenografts in mice.05. for caspase-8. tumor volume changes after 3-week treatment with 10 mg/kg per week docetaxel and/or 30 mg/kg per day PXD101 (PXD) for 5 days per week. .2562 HISTONE DEACETYLASE INHIBITOR POTENTIATES DOCETAXEL IN PROSTATE CANCER Figure 4.11.3% tumor volume in mice treated with 10 mg/kg per week docetaxel.

68: 917. A). and using tubulin and Bcl-2 family proteins as biomarkers in prostate cancer cell lines. Tannock IF. 8. an enzyme that acetylates cytosolic nonhistone proteins. 3. 5. However. These results suggest that pan-HDACIs may be used for hormone dependent and independent prostate cancer. Docetaxel stabilizes polymerized tubulin and increased tubulin acetylation is associated with such stabilization. 351: 1502. 2. Biochem Pharmacol 2007. Korea. The panHDACIs SAHA and PXD101 inhibited the growth of hormone dependent and hormone independent cells in a dose dependent manner (fig. Ann Oncol 2010. HDACIs enhance tubulin acetylation by inhibiting HDAC 6.7 In accordance with these reports we found that the docetaxel and HDACI combination increased the acetylated tubulin level compared with that using either reagent alone (fig. as shown by p21 expression assays. Nat Rev Cancer 2006. 26: 5541. N Engl J Med 2004. we tested the effects of combining docetaxel and PXD101 on tumor growth in nude mice injected with DU145 cells. D to I). 2). Western blotting for caspase showed that combination treatment activated extrinsic and intrinsic apoptosis pathways (fig. 59: 225.28. Although HDACI cytotoxicity was lower in PC3 than in DU145 cells. Jemal A. Xu WS.20. Siegel R. B). 351: 1513. ACKNOWLEDGMENTS SAHA and PXD101 were synthesized at Crystal Genomics. Our results show that this combination was also effective in vivo against HRPC cells (fig. Mol Pharmacol 2005. Acharya MR. 74: 659. Oncogene 2007. A). the cell line most sensitive to HDACIs of the 3 lines tested. 1. a sequential combination of docetaxel and HDACIs resulted in a synergistic increase in the antiproliferative effects of either drug given alone in DU145 HRPC cells in vitro and in vivo. docetaxel ¡HDACI combination treatment had a potent antiproliferative effect in PC3 cells (data not shown).29 We have extended these findings by noting that SAHA or PXD101 potentiated docetaxel toxicity. 9. A).HISTONE DEACETYLASE INHIBITOR POTENTIATES DOCETAXEL IN PROSTATE CANCER 2563 uated the antitumor effects of HDACIs on hormone dependent (LNCaP) and hormone independent (DU145 and PC3) prostate cancer cells. C). N Engl J Med 2004. 4. Using DU145 cells. 3. further study is needed to elucidate the influence of the combination schedule on the response. These results suggest that sequential treatment with docetaxel followed by SAHA or PXD101 may be a useful therapy for HRPC. Glaser KB: HDAC inhibitors: clinical update and mechanism-based potential. 4. Minucci S and Pelicci PG: Histone deacetylase inhibitors and the promise of epigenetic (and more) treatments for cancer. These preclinical findings support the clinical evaluation of HDACIs combined with docetaxel for HRPC. Venitz J et al: Rational development of histone deacetylase inhibitors as anticancer agents: a review. 4: 13. Petrylak DP. To elucidate other mechanisms of cell death we analyzed caspase activation and Bcl-2 family protein levels. Cell cycle analysis and Western blotting for caspase revealed that HDACIs induced prostate cancer cell apoptosis (fig. CONCLUSIONS Results indicate that HDACI can be used to treat patients with hormone independent and hormone dependent prostate cancer. 4. and decreased the anti-apoptotic proteins Mcl-1 and Bcl-xl (fig. 6. 3. median effect analysis and isobolograms indicated that the docetaxel and HDACI combination had synergy (fig. Since HDACIs are thought to act by potentiating the antitumor effects of chemotherapeutic agents and/or radiation. de Wit R. Because it was reported that combining SAHA plus docetaxel is poorly tolerated in HRPC cases30 and PXD101 is not a substrate of the multidrug resistance gene. an agent widely used for HRPC. Moreover. 7. Hussain MH et al: Docetaxel and estramustine compared with mitoxantrone and prednisone for advanced refractory prostate cancer. 2009. A). Depsipeptide (FK228) potentiated docetaxel toxicity in HRPC cells. 6: 38. Tangen CM. Johnstone RW and Licht JD: Histone deacetylase inhibitors in cancer therapy: is transcription the primary target? Cancer Cell 2003. 4. Berry WR et al: Docetaxel plus prednisone or mitoxantrone plus prednisone for advanced prostate cancer. .27 Also. Bik and Bim. Since all of our in vitro experiments indicated that combination treatment with docetaxel and HDACIs has an antitumor effect on HRPC cells by inducing apoptosis. Epub ahead of print. Seoul. Galsky MD and Vogelzang NJ: Docetaxel-based combination therapy for castration-resistant prostate cancer. Ward E et al: Cancer statistics. combination treatment increased the proapoptotic proteins t-Bid. 5. we chose the combination of PXD101 plus docetaxel. Also. we tested whether SAHA and PXD101 could potentiate the activity of docetaxel. we found that sequential treatment with docetaxel followed by HDACI (docetaxel ¡SAHA or PXD101) resulted in the greatest growth inhibition (fig.24 –26 and they showed cytotoxicity as monotreatment in HRPC cells in our study. CA Cancer J Clin 2009. Moreover. Sparreboom A. Parmigiani RB and Marks PA: Histone deacetylase inhibitors: molecular mechanisms of action. REFERENCES 1.

Thorne S. Monestiroli S. Schneider BJ. Fossa SD et al: Combination of bevacizumab and docetaxel in docetaxel-pretreated hormone-refractory prostate cancer: a phase 2 study. enhance the effects of gemcitabine and docetaxel in hormone refractory prostate cancer cells. 11. 17: 761. Ara G. 66: 181. 280: 26729. Daugaard G. 3: 300. 2009. Smith DC et al: Phase I study of vorinostat plus docetaxel in patients with solid tumor malignancies. 24. 100: 4389. suppl. 25: 261. Stanfield J. Int J Oncol 2004. 27: 15s. 70: 396. 104: 289. Curr Opin Pharmacol 2008. Rotili D et al: Histone deacetylation in epigenetics: an attractive target for anticancer therapy. Kumazawa T et al: Low concentrations of the histone deacetylase inhibitor. Buschbeck M. Wang X et al: Antitumor activity of the histone deacetylase inhibitor MS-275 in prostate cancer models. Zhang Z.2564 HISTONE DEACETYLASE INHIBITOR POTENTIATES DOCETAXEL IN PROSTATE CANCER 10. Eur Urol 2008.. Haggarty SJ. Bradley D. Sawyers CL and Scher HI: Targeting the androgen receptor pathway in prostate cancer. Cancer Res 2003. 21. 102: 4842. 25. 295: 85. in combination with docetaxel. 8: 440. 30. 13. Baek JH et al: Inhibition of histone deacetylase increases cytotoxicity to anticancer drugs targeting DNA. 28: 15s. suppl. Cancer Lett 2010. J Clin Oncol. Huebner G et al: An openlabel randomized phase II trial of belinostat (PXD101) in combination with carboplatin and paclitaxel (BelCaP) compared to carboplatin and paclitaxel in patients with previously untreated carcinoma of unknown primary. Abbas A and Gupta S: The role of histone deacetylases in prostate cancer. Schweyer S. Ross RW et al: A phase I study of oral panobinostat alone and in combination with docetaxel in patients with castrationresistant prostate cancer. Frenkel E et al: Enhanced therapeutic effect on androgen-independent prostate cancer by depsipeptide (FK228). depsipeptide. Hedjran F et al: Impact of the histone deacetylase inhibitors on taxotere resistance due to P-gp (MDR) and tubulin acetylation. a histone deacetylase inhibitor. Mills E et al: Activity of the histone deacetylase inhibitor belinostat (PXD101) in preclinical models of prostate cancer. Kakinuma H. Di Lorenzo G.. Prostate 2007. Wong JC et al: Domain-selective small-molecule inhibitor of histone deacetylase 6 (HDAC6)-mediated tubulin deacetylation. Bian X et al: Ku70 acetylation mediates neuroblastoma cell death induced by histone deacetylase inhibitors. J Biol Chem 2005. Koeller KM. 46. 15. Massa S. Proc Natl Acad Sci U S A 2005. Urology 2007. Fizazi K. Fontanella B. 2010. Subramanian C. Pranpat M. Bali P. 28. 27. Proc Amer Assoc Cancer Res 2005. 22. Kanzaki M. abstract 2528. Figg WD. 14. Oncol Rep 2007. 3: 831. 20. Mai A. 17. Piperno G. Cancer Biol Ther 2004. abstract TPS185. Qian DZ. Rathkopf D. Kim MS. 24: 25. Chen Y. 16. Reid T. 26. Qian X. D’Acunto CW. J Cell Biol 1987. Gutierrez A et al: Altered epigenetic signals in human disease. Opipari AW Jr. 67: 1182. Thelen P. Wei YF. 18. LeDizet M and Chang XJ: Microtubules containing acetylated alpha-tubulin in mammalian cells in culture. 63: 7291. 19. J Clin Oncol. Hemmerlein B et al: Expressional changes after histone deacetylase inhibition by valproic acid in LNCaP human prostate cancer cells. 122: 1400. Blake M. Epigenetics 2008. 23. Di Croce L. 54: 1089. Wong BY. 12. 29. Nat Med 2005. Proc Natl Acad Sci U S A 2003. Int J Cancer 2008. Rodriquez M et al: Histone deacetylase inhibitor FR235222 sensitizes human prostate adenocarcinoma cells to apoptosis through up-regulation of Annexin A1. Med Res Rev 2005. Bradner J et al: Inhibition of histone deacetylase 6 acetylates and disrupts the chaperone function of heat shock protein 90: a novel basis for antileukemia activity of histone deacetylase inhibitors. 11: 71. Insinga A. Ronzoni S et al: Inhibitors of histone deacetylases induce tumor-selective apoptosis through activation of the death receptor pathway. . Cancer Chemother Pharmacol 2010.