Small-molecule screening of PC3 prostate cancer cells identifies tilorone dihydrochloride to selectively inhibit cell growth based on cyclin-dependent kinase 5 expression

Wissing,Michel D.; Dadon,Tikva; Kim,Eunice; Piontek,Klaus B.; Shim,Joong S.; Kaelber,Nadine S.; Liu,Jun O.; Kachhap,Sushant K.; Nelkin,Barry D.

doi:10.3892/or.2014.3174

Small-molecule screening of PC3 prostate cancer cells identifies tilorone dihydrochloride to selectively inhibit cell growth based on cyclin-dependent kinase 5 expression

Authors:
- Michel D. Wissing
- Tikva Dadon
- Eunice Kim
- Klaus B. Piontek
- Joong S. Shim
- Nadine S. Kaelber
- Jun O. Liu
- Sushant K. Kachhap
- Barry D. Nelkin
View Affiliations

Affiliations: Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA, Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
Published online on: May 13, 2014 https://doi.org/10.3892/or.2014.3174
Pages: 419-424

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Cyclin-dependent kinase 5 (CDK5) is a potential target for prostate cancer treatment, the enzyme being essential for prostate tumor growth and formation of metastases. In the present study, we identified agents that target prostate cancer cells based on CDK5 expression. CDK5 activity was suppressed by transfection of PC3 prostate cancer cells with a dominant-negative construct (PC3 CDK5dn). PC3 CDK5dn and PC3 control cells were screened for compounds that selectively target cells based on CDK5 expression, utilizing the Johns Hopkins Drug Library. MTS proliferation, clonogenic and 3D growth assays were performed to validate the selected hits. Screening of 3,360 compounds identified rutilantin, ethacridine lactate and cetalkonium chloride as compounds that selectively target PC3 control cells and a tilorone analog as a selective inhibitor of PC3 CDK5dn cells. A PubMed literature study indicated that tilorone may have clinical use in patients. Validation experiments confirmed that tilorone treatment resulted in decreased PC3 cell growth and invasion; PC3 cells with inactive CDK5 were inhibited more effectively. Future studies are needed to unravel the mechanism of action of tilorone in CDK5 deficient prostate cancer cells and to test combination therapies with tilorone and a CDK5 inhibitor for its potential use in clinical practice.

View Figures

View References

1	Siegel R, Ma J, Zou Z and Jemal A: Cancer statistics, 2014. CA Cancer J Clin. 64:9–29. 2014. View Article : Google Scholar
2	Tarricone C, Dhavan R, Peng J, Areces LB, Tsai LH and Musacchio A: Structure and regulation of the CDK5-p25(nck5a) complex. Mol Cell. 8:657–669. 2001. View Article : Google Scholar : PubMed/NCBI
3	Rosales JL and Lee KY: Extraneuronal roles of cyclin-dependent kinase 5. Bioessays. 28:1023–1034. 2006. View Article : Google Scholar : PubMed/NCBI
4	Lalioti V, Pulido D and Sandoval IV: Cdk5, the multifunctional surveyor. Cell Cycle. 9:284–311. 2010. View Article : Google Scholar : PubMed/NCBI
5	Dhavan R and Tsai LH: A decade of CDK5. Nat Rev Mol Cell Biol. 2:749–759. 2001. View Article : Google Scholar : PubMed/NCBI
6	Cicero S and Herrup K: Cyclin-dependent kinase 5 is essential for neuronal cell cycle arrest and differentiation. J Neurosci. 25:9658–9668. 2005. View Article : Google Scholar : PubMed/NCBI
7	Strock CJ, Park JI, Nakakura EK, Bova GS, Isaacs JT, Ball DW and Nelkin BD: Cyclin-dependent kinase 5 activity controls cell motility and metastatic potential of prostate cancer cells. Cancer Res. 66:7509–7515. 2006. View Article : Google Scholar : PubMed/NCBI
8	Feldmann G, Mishra A, Hong SM, et al: Inhibiting the cyclin-dependent kinase CDK5 blocks pancreatic cancer formation and progression through the suppression of Ras-Ral signaling. Cancer Res. 70:7509–7515. 2010. View Article : Google Scholar : PubMed/NCBI
9	Hsu FN, Chen MC, Chiang MC, et al: Regulation of androgen receptor and prostate cancer growth by cyclin-dependent kinase 5. J Biol Chem. 286:33141–33149. 2011. View Article : Google Scholar : PubMed/NCBI
10	Demelash A, Rudrabhatla P, Pant HC, et al: Achaete-scute homologue-1 (ASH1) stimulates migration of lung cancer cells through Cdk5/p35 pathway. Mol Biol Cell. 23:2856–2866. 2012. View Article : Google Scholar : PubMed/NCBI
11	Hsu FN, Chen MC, Lin KC, et al: Cyclin-dependent kinase 5 modulates STAT3 and androgen receptor activation through phosphorylation of Ser⁷²⁷on STAT3 in prostate cancer cells. Am J Physiol Endocrinol Metab. 305:E975–E986. 2013. View Article : Google Scholar : PubMed/NCBI
12	Pozo K, Castro-Rivera E, Tan C, et al: The role of Cdk5 in neuroendocrine thyroid cancer. Cancer Cell. 24:499–511. 2013. View Article : Google Scholar : PubMed/NCBI
13	Chong CR, Qian DZ, Pan F, Wei Y, Pili R, Sullivan DJ and Liu JO: Identification of type 1 inosine monophosphate dehydrogenase as an antiangiogenic drug target. J Med Chem. 49:2677–2680. 2006. View Article : Google Scholar : PubMed/NCBI
14	Chong CR, Xu J, Lu J, Bhat S, Sullivan DJ and Liu JO: Inhibition of angiogenesis by the antifungal drug itraconazole. ACS Chem Biol. 2:263–270. 2007. View Article : Google Scholar : PubMed/NCBI
15	Rudin CM, Liu W, Desai A, et al: Pharmacogenomic and pharmacokinetic determinants of erlotinib toxicity. J Clin Oncol. 26:1119–1127. 2008. View Article : Google Scholar : PubMed/NCBI
16	Zhang H, Qian DZ, Tan YS, et al: Digoxin and other cardiac glycosides inhibit HIF-1alpha synthesis and block tumor growth. Proc Natl Acad Sci USA. 105:19579–19586. 2008. View Article : Google Scholar : PubMed/NCBI
17	Wissing MD, Mendonca J, Kim E, et al: Identification of cetrimonium bromide and irinotecan as compounds with synthetic lethality against NDRG1 deficient prostate cancer cells. Cancer Biol Ther. 14:401–410. 2013. View Article : Google Scholar : PubMed/NCBI
18	Nikolic M, Dudek H, Kwon YT, Ramos YF and Tsai LH: The cdk5/p35 kinase is essential for neurite outgrowth during neuronal differentiation. Genes Dev. 10:816–825. 1996. View Article : Google Scholar : PubMed/NCBI
19	Kachhap SK, Rosmus N, Collis SJ, et al: Downregulation of homologous recombination DNA repair genes by HDAC inhibition in prostate cancer is mediated through the E2F1 transcription factor. PLoS One. 5:e112082010. View Article : Google Scholar : PubMed/NCBI
20	O’Meara S, Al-Kurdi D, Ologun Y and Ovington LG: Antibiotics and antiseptics for venous leg ulcers. Cochrane Database Syst Rev. CD0035572010.PubMed/NCBI
21	Hou SP, Fang AH, Chen QF, Huang YM, Chen OJ and Cheng LN: Termination of second-trimester pregnancy by mifepristone combined with misoprostol versus intra-amniotic injection of ethacridine lactate (Rivanol^®): a systematic review of Chinese trials. Contraception. 84:214–223. 2011. View Article : Google Scholar : PubMed/NCBI
22	Daull P, Lallemand F and Garrigue JS: Benefits of cetalkonium chloride cationic oil-in-water nanoemulsions for topical ophthalmic drug delivery. J Pharm Pharmacol. May 26–2013.(Epub ahead of print). View Article : Google Scholar
23	Hume V, Westwood JC and Appleyard G: The anti-viral action of Rutilantin A. J Gen Microbiol. 38:143–151. 1965. View Article : Google Scholar : PubMed/NCBI
24	Krueger RE and Mayer GD: Tilorone hydrochloride: an orally active antiviral agent. Science. 169:1213–1214. 1970. View Article : Google Scholar : PubMed/NCBI
25	Mayer GD and Krueger RF: Tilorone hydrochloride: mode of action. Science. 169:1214–1215. 1970. View Article : Google Scholar : PubMed/NCBI
26	Tazulakhova EB, Parshina OV, Guseva TS and Ershov FI: Russian experience in screening, analysis, and clinical application of novel interferon inducers. J Interferon Cytokine Res. 21:65–73. 2001. View Article : Google Scholar : PubMed/NCBI
27	Adamson RH: Antitumor activity of tilorone hydrochloride against some rodent tumors: preliminary report. J Natl Cancer Inst. 46:431–434. 1971.PubMed/NCBI
28	Cummings FJ, Gelman R, Skeel RT, Kuperminc M, Israel L, Colsky J and Tormey D: Phase II trials of Baker’s antifol, bleomycin, CCNU, streptozotocin, tilorone, and 5-fluorodeoxyuridine plus arabinosyl cytosine in metastatic breast cancer. Cancer. 48:681–685. 1981.
29	Chong CR and Sullivan DJ: New uses for old drugs. Nature. 448:645–646. 2007. View Article : Google Scholar : PubMed/NCBI
30	Shim JS, Matsui Y, Bhat S, et al: Effect of nitroxoline on angiogenesis and growth of human bladder cancer. J Natl Cancer Inst. 102:1855–1873. 2010. View Article : Google Scholar : PubMed/NCBI
31	Shim JS, Rao R, Beebe K, Neckers L, Han I, Nahta R and Liu JO: Selective inhibition of HER2-positive breast cancer cells by the HIV protease inhibitor nelfinavir. J Natl Cancer Inst. 104:1576–1590. 2012. View Article : Google Scholar : PubMed/NCBI
32	Yang HC, Xing S, Shan L, et al: Small-molecule screening using a human primary cell model of HIV latency identifies compounds that reverse latency without cellular activation. J Clin Invest. 119:3473–3486. 2009.PubMed/NCBI
33	Lisianyi MI and Skitiak SA: Use of amiksin in complex therapy of cerebral gliomas. Lik Sprava. 121–123. 2002.(In Ukranian).
34	Karimova FS, Ivanchenko GF and Grigorian SS: The treatment of laryngeal papillomatosis with interferon inducers. Vestn Otorinolaringol. 54–57. 2000.(In Russian).
35	Zhou D, Tuo W, Hu H, et al: Synthesis and activity evaluation of tilorone analogs as potential anticancer agents. Eur J Med Chem. 64:432–441. 2013. View Article : Google Scholar : PubMed/NCBI
36	Ratan RR, Siddiq A, Aminova L, et al: Small molecule activation of adaptive gene expression: tilorone or its analogs are novel potent activators of hypoxia inducible factor-1 that provide prophylaxis against stroke and spinal cord injury. Ann NY Acad Sci. 1147:383–394. 2008. View Article : Google Scholar
37	Schrimpf MR, Sippy KB, Briggs CA, et al: SAR of α7 nicotinic receptor agonists derived from tilorone: exploration of a novel nicotinic pharmacophore. Bioorg Med Chem Lett. 22:1633–1638. 2012.