OBJECTIVES: Therapeutic potential of conventionally used platinum-based drugs in treatment of colorectal tumours has been limited due to high incidence of tumour resistance to them and to their severe side effects. This evokes a search for more suitable anti-cancer drugs. We have compared ability of oxaliplatin and a novel platinum(IV) complex, LA-12, to modulate the cell cycle and induce apoptosis in human colon adenocarcinoma HCT116 wt and p53/p21 null cells, and have investigated molecular mechanisms involved. MATERIALS AND METHODS: Cell cycle-related changes were analysed by flow cytometry (bromodeoxyuridine/propidium iodide staining, histone H3 phosphorylation). Apoptosis was detected using flow cytometry (assays monitoring caspase activity) and fluorescence microscopy (nuclear morphology). Changes in levels of genes/proteins involved in cell cycle and apoptosis regulation were examined by RT-PCR and western blotting. RESULTS: Our results highlight the outstanding ability of LA-12 to induce effective elimination of colon cancer cells independently of p53/p21, and in significantly lower doses compared to oxaliplatin. While oxaliplatin induced p53- and p21-dependent G2 -phase arrest associated with downregulation of cyclin B1 and Cdk1, LA-12 allowed cells to enter M-phase of the cell cycle regardless of p53/p21 status. CONCLUSIONS: Higher malignant cell toxicity and ability to bypass cell cycle arrest important for the cell damage repair suggest LA-12 to be a more effective candidate for elimination of colon tumours from a variety of genetic backgrounds, compared with oxaliplatin.

Department of Cytokinetics Institute of Biophysics Academy of Sciences of the Czech Republic v v i Brno 61265 Czech Republic; Department of Animal Physiology and Immunology Faculty of Science Institute of Experimental Biology Masaryk University Brno 621 00 Czech Republic

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Kelland L (2007) The resurgence of platinum‐based cancer chemotherapy. Nat. Rev. Cancer 7, 573–584. PubMed

Bastos DA, Ribeiro SC, De Freitas D, Hoff PM (2010) Combination therapy in high‐risk stage II or stage III colon cancer: current practice and future prospects. Ther. Adv. Med. Oncol. 2, 261–272. PubMed PMC

Douillard J‐Y, Bennouna J, Senellart H (2008) Is XELOX equivalent to FOLFOX or other continuous‐infusion 5‐fluorouracil chemotherapy in metastatic colorectal cancer? Clin. Colorectal Cancer 7, 206–211. PubMed

Choy H, Park C, Yao M (2008) Current status and future prospects for satraplatin, an oral platinum analogue. Clin. Cancer Res. 14, 1633–1638. PubMed

Zák F, Turánek J, Kroutil A, Sova P, Mistr A, Poulová A et al (2004) Platinum(IV) complex with adamantylamine as nonleaving amine group: synthesis, characterization, and in vitro antitumor activity against a panel of cisplatin‐resistant cancer cell lines. J. Med. Chem. 47, 761–763. PubMed

Kozubík A, Horváth V, Svihálková‐Sindlerová L, Soucek K, Hofmanová J, Sova P et al (2005) High effectiveness of platinum(IV) complex with adamantylamine in overcoming resistance to cisplatin and suppressing proliferation of ovarian cancer cells in vitro. Biochem. Pharmacol. 69, 373–383. PubMed

Horváth V, Blanárová O, Svihálková‐Sindlerová L, Soucek K, Hofmanová J, Sova P et al (2006) Platinum(IV) complex with adamantylamine overcomes intrinsic resistance to cisplatin in ovarian cancer cells. Gynecol. Oncol. 102, 32–40. PubMed

Svihálková‐Sindlerová L, Foltinová V, Vaculová A, Horváth V, Soucek K, Sova P et al (2010) LA‐12 overcomes confluence‐dependent resistance of HT‐29 colon cancer cells to Pt (II) compounds. Anticancer Res. 30, 1183–1188. PubMed

Sova P, Mistr A, Kroutil A, Zak F, Pouckova P, Zadinova M (2005) Preclinical anti‐tumor activity of a new oral platinum(IV) drug LA‐12. Anticancer Drugs 16, 653–657. PubMed

Sova P, Mistr A, Kroutil A, Zak F, Pouckova P, Zadinova M (2006) Comparative anti‐tumor efficacy of two orally administered platinum(IV) drugs in nude mice bearing human tumor xenografts. Anticancer Drugs 17, 201–206. PubMed

Sova P, Chladek J, Zak F, Mistr A, Kroutil A, Semerad M et al (2005) Pharmacokinetics and tissue distribution of platinum in rats following single and multiple oral doses of LA‐12 [(OC‐6‐43)‐bis(acetato)(1‐adamantylamine)amminedichloroplatinum(IV)]. Int. J. Pharm. 288, 123–129. PubMed

Sova P, Mistr A, Kroutil A, Semerád M, Chlubnová H, Hrusková V et al (2011) A comparative study of pharmacokinetics, urinary excretion and tissue distribution of platinum in rats following a single‐dose oral administration of two platinum(IV) complexes LA‐12 (OC‐6‐43)‐bis(acetato)(1‐adamantylamine)amminedichloroplatinum(IV) and satraplatin (OC‐6‐43)‐bis(acetato)amminedichloro(cyclohexylamine)platinum(IV). Cancer Chemother. Pharmacol. 67, 1247–1256. PubMed

Kaspárková J, Nováková O, Vrána O, Intini F, Natile G, Brabec V (2006) Molecular aspects of antitumor effects of a new platinum(IV) drug. Mol. Pharmacol. 70, 1708–1719. PubMed

Kvardova V, Hrstka R, Walerych D, Muller P, Matoulkova E, Hruskova V et al (2010) The new platinum(IV) derivative LA‐12 shows stronger inhibitory effect on Hsp90 function compared to cisplatin. Mol. Cancer 9, 147. PubMed PMC

Siddik ZH (2003) Cisplatin: mode of cytotoxic action and molecular basis of resistance. Oncogene 22, 7265–7279. PubMed

Niida H, Nakanishi M (2006) DNA damage checkpoints in mammals. Mutagenesis 21, 3–9. PubMed

Yu J, Zhang L (2005) The transcriptional targets of p53 in apoptosis control. Biochem. Biophys. Res. Commun. 331, 851–858. PubMed

Green DR, Kroemer G (2009) Cytoplasmic functions of the tumour suppressor p53. Nature 458, 1127–1130. PubMed PMC

Murray‐Zmijewski F, Slee EA, Lu X (2008) A complex barcode underlies the heterogeneous response of p53 to stress. Nat. Rev. Mol. Cell Biol. 9, 702–712. PubMed

Broude EV, Swift ME, Vivo C, Chang B‐D, Davis BM, Kalurupalle S et al (2007) p21(Waf1/Cip1/Sdi1) mediates retinoblastoma protein degradation. Oncogene 26, 6954–6958. PubMed

Coqueret O (2003) New roles for p21 and p27 cell‐cycle inhibitors: a function for each cell compartment? Trends Cell Biol. 13, 65–70. PubMed

Bunz F, Dutriaux A, Lengauer C, Waldman T, Zhou S, Brown JP et al (1998) Requirement for p53 and p21 to sustain G2 arrest after DNA damage. Science 282, 1497–1501. PubMed

Innocente SA, Lee JM (2005) p53 is a NF‐Y‐ and p21‐independent, Sp1‐dependent repressor of cyclin B1 transcription. FEBS Lett. 579, 1001–1007. PubMed

Basile V, Mantovani R, Imbriano C (2006) DNA damage promotes histone deacetylase 4 nuclear localization and repression of G2/M promoters, via p53 C‐terminal lysines. J. Biol. Chem. 281, 2347–2357. PubMed

Müller GA, Engeland K (2010) The central role of CDE/CHR promoter elements in the regulation of cell cycle‐dependent gene transcription. FEBS J. 277, 877–893. PubMed

Waldman T, Kinzler KW, Vogelstein B (1995) p21 is necessary for the p53‐mediated G1 arrest in human cancer cells. Cancer Res. 55, 5187–5190. PubMed

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real‐time quantitative PCR and the 2(‐Delta Delta C(T)) Method. Methods 25, 402–408. PubMed

Juan G, Traganos F, Darzynkiewicz Z (1999) Histone H3 phosphorylation in human monocytes and during HL‐60 cell differentiation. Exp. Cell Res. 246, 212–220. PubMed

Misset JL, Bleiberg H, Sutherland W, Bekradda M, Cvitkovic E (2000) Oxaliplatin clinical activity: a review. Crit. Rev. Oncol. Hematol. 35, 75–93. PubMed

Liu YP, Ling Y, Qi QF, Zhang YP, Zhang CS, Zhu CT et al (2013) The effects of ERCC1 expression levels on the chemosensitivity of gastric cancer cells to platinum agents and survival in gastric cancer patients treated with oxaliplatin‐based adjuvant chemotherapy. Oncol. Lett. 5, 935–942. PubMed PMC

McWhinney SR, Goldberg RM, McLeod HL (2009) Platinum neurotoxicity pharmacogenetics. Mol. Cancer Ther. 8, 10–16. PubMed PMC

Hata T, Yamamoto H, Ngan CY, Koi M, Takagi A, Damdinsuren B et al (2005) Role of p21waf1/cip1 in effects of oxaliplatin in colorectal cancer cells. Mol. Cancer Ther. 4, 1585–1594. PubMed

Arango D, Wilson AJ, Shi Q, Corner GA, Arañes MJ, Nicholas C et al (2004) Molecular mechanisms of action and prediction of response to oxaliplatin in colorectal cancer cells. Br. J. Cancer 91, 1931–1946. PubMed PMC

Hayward RL, Macpherson JS, Cummings J, Monia BP, Smyth JF, Jodrell DI (2004) Enhanced oxaliplatin‐induced apoptosis following antisense Bcl‐xl down‐regulation is p53 and Bax dependent: genetic evidence for specificity of the antisense effect. Mol. Cancer Ther. 3, 169–178. PubMed

Chiu S‐J, Hsu T‐S, Chao J‐I (2006) Opposing securin and p53 protein expression in the oxaliplatin‐induced cytotoxicity of human colorectal cancer cells. Toxicol. Lett. 167, 122–130. PubMed

Rakitina TV, Vasilevskaya IA, O'Dwyer PJ (2007) Inhibition of G1/S transition potentiates oxaliplatin‐induced cell death in colon cancer cell lines. Biochem. Pharmacol. 73, 1715–1726. PubMed

Toscano F, Parmentier B, Fajoui ZE, Estornes Y, Chayvialle J‐A, Saurin J‐C et al (2007) p53 dependent and independent sensitivity to oxaliplatin of colon cancer cells. Biochem. Pharmacol. 74, 392–406. PubMed

Zaanan A, Cuilliere‐Dartigues P, Guilloux A, Parc Y, Louvet C, De Gramont A et al (2010) Impact of p53 expression and microsatellite instability on stage III colon cancer disease‐free survival in patients treated by 5‐fluorouracil and leucovorin with or without oxaliplatin. Ann. Oncol. 21, 772–780. PubMed

Hrstka R, Powell DJ, Kvardova V, Roubalova E, Bourougaa K, Candeias MM et al (2008) The novel platinum(IV) complex LA‐12 induces p53 and p53/47 responses that differ from the related drug, cisplatin. Anticancer Drugs 19, 369–379. PubMed

Roubalová E, Kvardová V, Hrstka R, Borilová S, Michalová E, Dubská L et al (2010) The effect of cellular environment and p53 status on the mode of action of the platinum derivative LA‐12. Invest. New Drugs 28, 445–453. PubMed

Kozubík A, Vaculová A, Soucek K, Vondrácek J, Turánek J, Hofmanová J (2008) Novel Anticancer Platinum(IV) Complexes with Adamantylamine: their Efficiency and Innovative Chemotherapy Strategies Modifying Lipid Metabolism. Met. Based Drugs 2008, 417897. PubMed PMC

Falck J, Mailand N, Syljuasen RG, Bartek J, Lukas J (2001) The ATM‐Chk2‐Cdc25A checkpoint pathway guards against radioresistant DNA synthesis. Nature 410, 842–847. PubMed

Castedo M, Perfettini JL, Roumier T, Kroemer G (2002) Cyclin‐dependent kinase‐1: linking apoptosis to cell cycle and mitotic catastrophe. Cell Death Differ. 9, 1287–1293. PubMed

Voland C, Bord A, Péleraux A, Pénarier G, Carrière D, Galiègue S et al (2006) Repression of cell cycle‐related proteins by oxaliplatin but not cisplatin in human colon cancer cells. Mol. Cancer Ther. 5, 2149–2157. PubMed

Chiu S‐J, Lee Y‐J, Hsu T‐S, Chen W‐S (2009) Oxaliplatin‐induced gamma‐H2AX activation via both p53‐dependent and ‐independent pathways but is not associated with cell cycle arrest in human colorectal cancer cells. Chem. Biol. Interact. 182, 173–182. PubMed

Fujie Y, Yamamoto H, Ngan CY, Takagi A, Hayashi T, Suzuki R et al (2005) Oxaliplatin, a potent inhibitor of survivin, enhances paclitaxel‐induced apoptosis and mitotic catastrophe in colon cancer cells. Jpn. J. Clin. Oncol. 35, 453–463. PubMed

William‐Faltaos S, Rouillard D, Lechat P, Bastian G (2006) Cell cycle arrest and apoptosis induced by oxaliplatin (L‐OHP) on four human cancer cell lines. Anticancer Res. 26, 2093–2099. PubMed

Horváth V, Soucek K, Svihálková‐Sindlerová L, Vondrácek J, Blanárová O, Hofmanová J et al (2007) Different cell cycle modulation following treatment of human ovarian carcinoma cells with a new platinum(IV) complex vs cisplatin. Invest. New Drugs 25, 435–443. PubMed

Russell P, Nurse P (1986) cdc25 +  functions as an inducer in the mitotic control of fission yeast. Cell 45, 145–153. PubMed

Russell P, Nurse P (1987) Negative regulation of mitosis by wee1 + , a gene encoding a protein kinase homolog. Cell 49, 559–567. PubMed

Cisplatin or LA-12 enhance killing effects of TRAIL in prostate cancer cells through Bid-dependent stimulation of mitochondrial apoptotic pathway but not caspase-10

Chk1 Inhibitor SCH900776 Effectively Potentiates the Cytotoxic Effects of Platinum-Based Chemotherapeutic Drugs in Human Colon Cancer Cells

Loss of PTEN Facilitates Rosiglitazone-Mediated Enhancement of Platinum(IV) Complex LA-12-Induced Apoptosis in Colon Cancer Cells