Skip to main content
SpringerLink
Log in

Polyamine depletion enhances the roscovitine-induced apoptosis through the activation of mitochondria in HCT116 colon carcinoma cells

  • Original Article
  • Published:
Amino Acids Aims and scope Submit manuscript

Abstract

Small molecule inhibitors of cyclin-dependent kinases (CDKs) show high therapeutic potential in various cancer types which are characterized by the accumulation of transformed cells due to impaired apoptotic machinery. Roscovitine, a CDK inhibitor showed to be a potent apoptotic inducer in several cancer cells. Polyamines, putrescine, spermidine and spermine, are biogenic amines involved in many cellular processes, including apoptosis. In this study, we explored the potential role of polyamines in roscovitine-induced apoptosis in HCT116 colon cancer cells. Roscovitine induced apoptosis by activating mitochondrial pathway caspases and modulating the expression of Bcl-2 family members. Depletion of polyamines by treatment with difluoromethylornithine (DFMO) increased roscovitine-induced apoptosis. Transient silencing of ornithine decarboxylase, polyamine biosynthesis enzyme and special target of DFMO also increased roscovitine-induced apoptosis in HCT116 cells. Interestingly, additional putrescine treatment was found pro-apoptotic due to the presence of non-functional ornithine decarboxylase (ODC). Finally, roscovitine altered polyamine catabolic pathway and led to decrease in putrescine and spermidine levels. Therefore, the metabolic regulation of polyamines may dictate the power of roscovitine induced apoptotic responses in HCT116 colon cancer cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

APAO:

N1-acetylpolyamine oxidase

CDKs:

Cyclin dependent kinases

CST:

Cell signaling technology

DFMO:

d,l-α-difluoromethylornithine

DMSO:

Dimethyl sulfoxide

H2-DCFDA:

2′-7′ Dichlorofluorescein

IEC-6:

Intestine epithelial cells of rats

MMP:

Mitochondrial membrane potential

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide

NAC:

N-acetyl cystein

ODC:

Ornithine decarboxylase

PAs:

Polyamines

PAO:

Polyamine oxidase

POD:

Peroxidase

Put:

Putrescine

SMO:

Spermine oxidase

ROS:

Reactive oxygen species

RT:

Reverse transcriptase

Spd:

Spermidine

Spm:

Spermine

SSAT:

Spermidine/spermine N1-acetyltransferase

5-FU:

5-Fluorouracil

References

  • Agostinelli E, Belli F, Molinari A, Condello M, Palmigiani P, Vedova LD, Marra M, Seiler N, Arancia G (2006) Toxicity of enzymatic oxidation products of spermine to human melanoma cells (M14): sensitization by heat and MDL 72527. Biochim Biophys Acta 1763:1040–1050

    Article  PubMed  CAS  Google Scholar 

  • Agostinelli E, Tempera G, Molinari A, Salvi M, Battaglia V, Toninello A, Arancia G (2007) The physiological role of biogenic amines redox reactions in mitochondria. New perspectives in cancer therapy. Amino acids 33:175–187

    Article  PubMed  CAS  Google Scholar 

  • Agostinelli E, Tempera G, Viceconte N, Saccoccio S, Battaglia V, Grancara S, Toninello A, Stevanato R (2010) Potential anticancer application of polyamine oxidation products formed by amine oxidase: a new therapeutic approach. Amino acids 38:353–368

    Article  PubMed  CAS  Google Scholar 

  • Aldoss IT, Tashi T, Ganti AK (2009) Seliciclib in malignancies. Expert Opin Investig Drugs 18:1957–1965

    Article  PubMed  CAS  Google Scholar 

  • Alvi AJ, Austen B, Weston VJ, Fegan C, MacCallum D, Gianella-Borradori A, Lane DP, Hubank M, Powell JE, Wei W, Taylor AM, Moss PA, Stankovic T (2005) A novel CDK inhibitor, CYC202 (R-roscovitine), overcomes the defect in p53-dependent apoptosis in B-CLL by down-regulation of genes involved in transcription regulation and survival. Blood 105:4484–4491

    Article  PubMed  CAS  Google Scholar 

  • Andersson G, Heby O (1977) Kinetics of cell proliferation and polyamine synthesis during Ehrlich ascites tumor growth. Cancer Res 37:4361–4366

    PubMed  CAS  Google Scholar 

  • Appleyard MV, O’Neill MA, Murray KE, Paulin FE, Bray SE, Kernohan NM, Levison DA, Lane DP, Thompson AM (2009) Seliciclib (CYC202, R-roscovitine) enhances the antitumor effect of doxorubicin in vivo in a breast cancer xenograft model. Int J Cancer 124:465–472

    Article  PubMed  CAS  Google Scholar 

  • Arisan ED, Kutuk O, Tezil T, Bodur C, Telci D, Basaga H (2010) Small inhibitor of Bcl-2, HA14–1, selectively enhanced the apoptotic effect of cisplatin by modulating Bcl-2 family members in MDA-MB-231 breast cancer cells. Breast Cancer Res Treat 119:271–281

    Article  PubMed  CAS  Google Scholar 

  • Bachrach U (1976) Polyamines as chemical markers of malignancy. The Italian J Biochem 25:77–93

    CAS  Google Scholar 

  • Bragado P, Armesilla A, Silva A, Porras A (2007) Apoptosis by cisplatin requires p53 mediated p38alpha MAPK activation through ROS generation. Apoptosis 12:1733–1742

    Article  PubMed  CAS  Google Scholar 

  • Burns MR, Graminski GF, Weeks RS, Chen Y, O’Brien TG (2009) Lipophilic lysine-spermine conjugates are potent polyamine transport inhibitors for use in combination with a polyamine biosynthesis inhibitor. J Med Chem 52:1983–1993

    Article  PubMed  CAS  Google Scholar 

  • Casero RA, Pegg AE (2009) Polyamine catabolism and disease. Biochem J 421:323–338

    Article  PubMed  CAS  Google Scholar 

  • Chen L, Jiang J, Cheng C, Yang A, He Q, Li D, Wang Z (2007) P53 dependent and independent apoptosis induced by lidamycin in human colorectal cancer cells. Cancer Biol Ther 6:965–973

    Article  PubMed  CAS  Google Scholar 

  • Chen Y, Weeks RS, Burns MR, Boorman DW, Klein-Szanto A, O’Brien TG (2006) Combination therapy with 2-difluoromethylornithine and a polyamine transport inhibitor against murine squamous cell carcinoma. Int J Cancer 118:2344–2349

    Article  PubMed  CAS  Google Scholar 

  • Cheok CF, Dey A, Lane DP (2007) Cyclin-dependent kinase inhibitors sensitize tumor cells to nutlin-induced apoptosis: a potent drug combination. Mol Cancer Res 5:1133–1145

    Article  PubMed  CAS  Google Scholar 

  • Choi SH, Kim SW, Choi DH, Min BH, Chun BG (2000) Polyamine-depletion induces p27Kip1 and enhances dexamethasone-induced G1 arrest and apoptosis in human T lymphoblastic leukemia cells. Leuk Res 24:119–127

    Article  PubMed  CAS  Google Scholar 

  • Choi W, Gerner EW, Ramdas L, Dupart J, Carew J, Proctor L, Huang P, Zhang W, Hamilton SR (2005) Combination of 5-fluorouracil and N1, N11-diethylnorspermine markedly activates spermidine/spermine N1-acetyltransferase expression, depletes polyamines, and synergistically induces apoptosis in colon carcinoma cells. J Biol Chem 280:3295–3304

    Article  PubMed  CAS  Google Scholar 

  • David-Pfeuty T (1999) Potent inhibitors of cyclin-dependent kinase 2 induce nuclear accumulation of wild-type p53 and nucleolar fragmentation in human untransformed and tumor-derived cells. Oncogene 18:7409–7422

    Article  PubMed  CAS  Google Scholar 

  • Evageliou NF, Hogarty MD (2009) Disrupting polyamine homeostasis as a therapeutic strategy for neuroblastoma. Clin Cancer Res 15:5956–5961

    Article  PubMed  CAS  Google Scholar 

  • Fischer SM, Conti CJ, Viner J, Aldaz CM, Lubet RA (2003) Celecoxib and difluoromethylornithine in combination have strong therapeutic activity against UV-induced skin tumors in mice. Carcinogenesis 24:945–952

    Article  PubMed  CAS  Google Scholar 

  • Garrofe-Ochoa X, Melero-Fernandez de Mera RM, Fernandez-Gomez FJ, Ribas J, Jordan J, Boix J (2008) BAX and BAK proteins are required for cyclin-dependent kinase inhibitory drugs to cause apoptosis. Mol Cancer Ther 7:3800–3806

    Article  PubMed  CAS  Google Scholar 

  • Hahntow IN, Schneller F, Oelsner M, Weick K, Ringshausen I, Fend F, Peschel C, Decker T (2004) Cyclin-dependent kinase inhibitor Roscovitine induces apoptosis in chronic lymphocytic leukemia cells. Leukemia 18:747–755

    Article  PubMed  CAS  Google Scholar 

  • Hallaert DY, Spijker R, Jak M, Derks IA, Alves NL, Wensveen FM, de Boer JP, de Jong D, Green SR, van Oers MH, Eldering E (2007) Crosstalk among Bcl-2 family members in B-CLL: seliciclib acts via the Mcl-1/Noxa axis and gradual exhaustion of Bcl-2 protection. Cell Death Differ 14:1958–1967

    Article  PubMed  CAS  Google Scholar 

  • He J, Fan M, Fang Q (2000) Difluoromethylornithine synergizes with antisense bcl-2 RNA in the induction of apoptosis of HL60 cells. Zhonghua Zhong Liu Za Zhi 22:105–108

    PubMed  CAS  Google Scholar 

  • Heby O, Andersson G, Gray JW (1978) Interference with S and G2 phase progression by polyamine synthesis inhibitors. Exp Cell Res 111:461–464

    Article  PubMed  CAS  Google Scholar 

  • Hudson EA, Howells LM, Gallacher-Horley B, Fox LH, Gescher A, Manson MM (2003) Growth-inhibitory effects of the chemopreventive agent indole-3-carbinol is increased in combination with the polyamine putrescine in the SW480 colon tumour cell line. BMC Cancer 3:2–20

    Article  PubMed  Google Scholar 

  • Hui AB, Yue S, Shi W, Alajez NM, Ito E, Green SR, Frame S, O’Sullivan B, Liu FF (2009) Therapeutic efficacy of seliciclib in combination with ionizing radiation for human nasopharyngeal carcinoma. Clin Cancer Res 15:3716–3724

    Article  PubMed  CAS  Google Scholar 

  • Iseki H, Ko TC, Xue XY, Seapan A, Hellmich MR, Townsend CM Jr (1997) Cyclin-dependent kinase inhibitors block proliferation of human gastric cancer cells. Surgery 122:187–194 (discussion 194–195)

    Google Scholar 

  • Kutuk O, Arisan ED, Tezil T, Shoshan MC, Basaga H (2009) Cisplatin overcomes Bcl-2-mediated resistance to apoptosis via preferential engagement of Bak: critical role of Noxa-mediated lipid peroxidation. Carcinogenesis 30:1517–1527

    Article  PubMed  CAS  Google Scholar 

  • Leitch AE, Riley NA, Sheldrake TA, Festa M, Fox S, Duffin R, Haslett C, Rossi AG (2010) The cyclin-dependent kinase inhibitor R-roscovitine down-regulates Mcl-1 to override pro-inflammatory signalling and drive neutrophil apoptosis. Eur J Immunol 40:1127–1138

    Article  PubMed  CAS  Google Scholar 

  • Marton LJ, Pegg AE (1995) Polyamines as targets for therapeutic intervention. Ann Rev Pharmac Toxicol 35:55–91

    Article  CAS  Google Scholar 

  • Marton LJ, Pegg AE, Morris DR (1991) Directions for polyamine research. J Cell Biochem 45:7–8

    Article  PubMed  CAS  Google Scholar 

  • Maurer M, Komina O, Wesierska-Gadek J (2009) Roscovitine differentially affects asynchronously growing and synchronized human MCF-7 breast cancer cells. Ann N Y Acad Sci 1171:250–256

    Article  PubMed  CAS  Google Scholar 

  • Meijer L, Borgne A, Mulner O, Chong JP, Blow JJ, Inagaki N, Inagaki M, Delcros JG, Moulinoux JP (1997) Biochemical and cellular effects of roscovitine, a potent and selective inhibitor of the cyclin-dependent kinases cdc2, cdk2 and cdk5. Eur J Biochem 243:527–536

    Article  PubMed  CAS  Google Scholar 

  • Mohapatra S, Chu B, Zhao X, Djeu J, Cheng JQ, Pledger WJ (2009) Apoptosis of metastatic prostate cancer cells by a combination of cyclin-dependent kinase and AKT inhibitors. Int J Biochem Cell Biol 41:595–602

    Article  PubMed  CAS  Google Scholar 

  • Monti MG, Pernecco L, Manfredini R, Frassineti C, Barbieri D, Marverti G, Ghiaroni S (1996) Inhibition of cell growth by accumulated spermine is associated with a transient alteration of cell cycle progression. Life Sci 58:2065–2072

    Article  PubMed  CAS  Google Scholar 

  • Odenlund M, Holmqvist B, Baldetorp B, Hellstrand P, Nilsson BO (2009) Polyamine synthesis inhibition induces S phase cell cycle arrest in vascular smooth muscle cells. Amino Acids 36:273–282

    Article  PubMed  CAS  Google Scholar 

  • Ortiz-Ferron G, Yerbes R, Eramo A, Lopez-Perez AI, De Maria R, Lopez-Rivas A (2008) Roscovitine sensitizes breast cancer cells to TRAIL-induced apoptosis through a pleiotropic mechanism. Cell Res 18:664–676

    Article  PubMed  CAS  Google Scholar 

  • Paprskarova M, Krystof V, Jorda R, Dzubak P, Hajduch M, Wesierska-Gadek J, Strnad M (2009) Functional p53 in cells contributes to the anticancer effect of the cyclin-dependent kinase inhibitor roscovitine. J Cell Biochem 107:428–437

    Article  PubMed  CAS  Google Scholar 

  • Schutte B, Nieland L, van Engeland M, Henfling ME, Meijer L, Ramaekers FC (1997) The effect of the cyclin-dependent kinase inhibitor olomoucine on cell cycle kinetics. Exp Cell Res 236:4–15

    Article  PubMed  CAS  Google Scholar 

  • Stanic I, Facchini A, Borzi RM, Vitellozzi R, Stefanelli C, Goldring MB, Guarnieri C, Flamigni F (2006) Polyamine depletion inhibits apoptosis following blocking of survival pathways in human chondrocytes stimulated by tumor necrosis factor-alpha. J Cell Physiol 206:138–146

    Article  PubMed  CAS  Google Scholar 

  • Wang X, Feith DJ, Welsh P, Coleman CS, Lopez C, Woster PM, O’Brien TG, Pegg AE (2007) Studies of the mechanism by which increased spermidine/spermine N1-acetyltransferase activity increases susceptibility to skin carcinogenesis. Carcinogenesis 28:2404–2411

    Article  PubMed  CAS  Google Scholar 

  • Wang Y, Casero RA Jr (2006) Mammalian polyamine catabolism: a therapeutic target, a pathological problem, or both? J Biochem 139:17–25

    Article  PubMed  CAS  Google Scholar 

  • Wesierska-Gadek J, Borza A, Komina O, Maurer M (2009) Impact of roscovitine, a selective CDK inhibitor, on cancer cells: bi-functionality increases its therapeutic potential. Acta Biochim Pol 56:495–501

    PubMed  CAS  Google Scholar 

  • Wesierska-Gadek J, Hajek SB, Sarg B, Wandl S, Walzi E, Lindner H (2008a) Pleiotropic effects of selective CDK inhibitors on human normal and cancer cells. Biochem Pharmacol 76:1503–1514

    Article  PubMed  CAS  Google Scholar 

  • Wesierska-Gadek J, Kramer MP, Maurer M (2008b) Resveratrol modulates roscovitine-mediated cell cycle arrest of human MCF-7 breast cancer cells. Food Chem Toxicol 46:1327–1333

    Article  PubMed  CAS  Google Scholar 

  • Xie SQ, Liu GC, Ma YF, Cheng PF, Wu YL, Wang MW, Ji BS, Zhao J, Wang CJ (2008) Synergistic antitumor effects of anthracenylmethyl homospermidine and alpha-difluoromethylornithine on promyelocytic leukemia HL60 cells. Toxicol In Vitro 22:352–358. doi:S0887-2333(07)00261-5

    Article  PubMed  CAS  Google Scholar 

  • Yuan Q, Ray RM, Johnson LR (2002) Polyamine depletion prevents camptothecin-induced apoptosis by inhibiting the release of Cytochrome c. Am J Physiol Cell Physiol 282:C1290–C1297

    PubMed  CAS  Google Scholar 

  • Zhang F, Zhang T, Gu ZP, Zhou YA, Han Y, Li XF, Wang XP, Cheng QS, Mei QB (2008) Enhancement of radiosensitivity by roscovitine pretreatment in human non-small cell lung cancer A549 cells. J Radiat Res (Tokyo) 49:541–548

    Article  CAS  Google Scholar 

  • Zhang F, Zhang T, Jiang T, Zhang R, Teng ZH, Li C, Gu ZP, Mei Q (2009) Wortmannin potentiates roscovitine-induced growth inhibition in human solid tumor cells by repressing PI3K/Akt pathway. Cancer Lett 286:232–239

    Article  PubMed  CAS  Google Scholar 

  • Zolnierczyk JD, Blonski JZ, Robak T, Kilianska ZM, Wesierska-Gadek J (2009) Roscovitine triggers apoptosis in B-cell chronic lymphocytic leukemia cells with similar efficiency as combinations of conventional purine analogs with cyclophosphamide. Ann N Y Acad Sci 1171:124–131

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was partially supported by Turkish Scientific and Technical Research Council Project, TUBITAK-TBAG 108T630 and Istanbul Kultur University Scientific Projects Support Center.

Author information

Authors and Affiliations

  1. Molecular Biology and Genetics Department, Istanbul Kultur University, Science and Literature Faculty, Atakoy Campus, 34156, Istanbul, Turkey

    Elif Damla Arısan, Ajda Çoker & Narçin Palavan-Ünsal

Authors
  1. Elif Damla Arısan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ajda Çoker
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Narçin Palavan-Ünsal
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Narçin Palavan-Ünsal.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Arısan, E.D., Çoker, A. & Palavan-Ünsal, N. Polyamine depletion enhances the roscovitine-induced apoptosis through the activation of mitochondria in HCT116 colon carcinoma cells. Amino Acids 42, 655–665 (2012). https://doi.org/10.1007/s00726-011-1040-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00726-011-1040-x

Keywords

Search

Navigation