Synergistic Anticancer Activity of Topotecan— Cyclin-Dependent Kinase Inhibitor Combinations against Drug-Resistant Small Cell Lung Cancer (SCLC) Cell Lines


Extended-stage small cell lung cancer (SCLC) responds to platinum/vepeside-based first-line chemotherapy but relapses rapidly as drug-resistant tumor. Topotecan (TPT) is the single chemotherapeutic agent approved for second-line treatment of SCLC. However, the response to TPT is short-lived and novel treatment modalities need to be developed. Sequential treatment of cytotoxic drugs and inhibitors of cyclin-dependent kinases (CDKs) showed promising preclinical anticancer activity and, in the present work, combinations of TPT with CDK inhibitors olomoucine, roscovitine and CDK4I are shown to exhibit synergistic cytotoxic activity against SCLC cell lines. Highest activity was found against TPT-resistant NCI-H417 and DMS153 cell lines and moderate chemosensitizing effects against a primary SCLC cell line and sensitive GLC19 cells at levels of CDK inhibitors which exerted low toxicity. A combination of 0.6 μM TPT with 0.6 μM roscovitine, exhibiting no significant cytotoxicity as single agents, reduced viability of the TPT-resistant NCI-H417 line (IC50 > 10 μM) by 50%. In the TPT resistant cell lines olomoucine and roscovitine, targeting CDK1,2,5,7, were highly effective, whereas in the more sensitive cell lines CDK4I, inhibiting mainly CDK4/6, showed activity. In NCI-417 cells, preincubation with roscovitine for one day proved synergistic with TPT. Thus, in good accordance with previous findings, CDK inhibitors are able to convert SCLC cancer cells which are cell-cycle arrested by a blockade of topoisomerase I by TPT to apoptotic cells. Since nowadays several CDK inhibitors are at various phases of clinical testing their combination with TPT seems to constitute a promising approach to improve second-line chemotherapy in SCLC.

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Hamilton, G. , Lukas Klameth, U. , Ulsperger, E. and Geissler, K. (2013) Synergistic Anticancer Activity of Topotecan— Cyclin-Dependent Kinase Inhibitor Combinations against Drug-Resistant Small Cell Lung Cancer (SCLC) Cell Lines. Journal of Cancer Therapy, 4, 47-53. doi: 10.4236/jct.2013.48A008.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] D. R. Youlden, S. M. Cramb and P. D. Baade, “The International Epidemiology of Lung Cancer: Geographical Distribution and Secular Trends,” Journal of Thoracic Oncology, Vol. 3, No. 8, 2008, pp. 819-831. doi:10.1097/JTO.0b013e31818020eb
[2] S. G. Spiro and G. A. Silvestri, “One Hundred Years of Lung Cancer,” American Journal of Respiratory Critical Care Medicine, Vol. 172, No. 5, 2005, pp. 523-529. doi:10.1164/rccm.200504-531OE
[3] A. Califano, Z. Abidin, R. Peck, C. Faivre-Finn and P. R. Lorigan, “Management of Small Cell Lung Cancer: Recent Developments for Optimal Care,” Drugs, Vol. 72, No. 4, 2012, pp. 471-490. doi:10. 2165/11597640-000000000-00000
[4] R. Govindan, N. Page, D. Morgensztern, et al., “Changing Epidemiology of Small-Cell Lung Cancer in the United States over the Last 30 Years: Analysis of the Surveillance, Epidemiologic, and End Results Database,” Journal of Clinical Oncology, Vol. 24, No. 28, 2006, pp. 4539-4544. doi:10.1200/JCO.2005.04.4859
[5] W. N. William and B. S. Glisson, “Novel Strategies for the Treatment of Small-Cell Lung Carcinoma,” Nature Reviews Clinical Oncology, Vol. 8, No. 10, 2011, pp. 611-619. doi:10.1038/nrclinonc.2011.90
[6] Y. Agra, M. Pelayo, M. Sacristan, et al., “Chemotherapy versus Best Supportive Care for Extensive Small Cell Lung Cancer,” Cochrane Database Systematic Review, Vol. 4, 2003, Article ID: CD001990.
[7] M. Takada, M. Fukuoka, M. Kawahara, et al., “Phase III Study of Concurrent versus Sequential Thoracic Radiotherapy in Combination with Cisplatin and Etoposide for Limited-Stage Small-Cell Lung Cancer,” Journal of Clinical Oncology, Vol. 20, No. 14, 2002, pp. 3054-3060. doi:10. 1200/JCO. 2002.12.071
[8] A. Rossi, M. Di Maio, P. Chiodini, et al., “Carboplatinor Cisplatin-Based Chemotherapy in First-Line Treatment of Small-Cell Lung Cancer: The COCIS Meta-Analysis of Individual Patient Data,” Journal of Clinical Oncology, Vol. 30, No. 14, 2012, pp. 1692-1698. doi:10.1200/JCO.2011.40.4905
[9] B. Fischer and A. Arcaro, “Current Status of Clinical Trials for Small Cell Lung Cancer,” Review of Recent Clinical Trials, Vol. 3, No. 1, 2008, pp. 40-61. doi:10.2174/157488708783330503
[10] D. Ormrod and C. M. Spencer, “Topotecan: A Review of Its Efficacy in Small Cell Lung Cancer,” Drugs, Vol. 58, No. 3, pp. 533-551. doi:10.2165/00003495-199958030-00020
[11] J. L. Hurwitz, F. McCoy, P. Scullin and D. A. Fennell, “New Advances in the Second-Line Treatment of Small Cell Lung Cancer,” Oncologist, Vol. 14, No. 10, 2009, pp. 986-994. doi:10. 1634/ theoncologist.2009-0026
[12] A. Schmittel, “Second-Line Therapy for Small-Cell Lung Cancer,” Expert Review in Anticancer Therapy, Vol. 11, No. 4, 2011, pp. 631-637. doi:10.1586/era.11.7
[13] M. T. Tomicic and K. B. aina, “Topoisomerase Degradation, DSB Repair, p53 and IAPs in Cancer Cell Resistance to Camptothecin-Like Topoisomerase I Inhibitors,” Biochimica et Biophysica Acta, Vol. 1835, No. 1, 2013, pp. 11-27.
[14] S. Agelaki, E. Kontopodis, A. Kotsakis, et al., “A Phase I Clinical Trial of Weekly Oral Topotecan for Relapsed Small Cell Lung Cancer,” Cancer Chemotherapy and Pharmacology, 2013, in press. doi:10.1007/s00280-013-2167-0
[15] R. Riemsma, J. P. Simons, Z. Bashir, C. L. Gooch and J. Kleijnen, “Systematic Review of Topotecan (Hycamtin) in Relapsed Small Cell Lung Cancer,” BMC Cancer, Vol. 10, 2010, p. 436.
[16] M. E. O’Brien, T. E.Ciuleanu, H. Tsekov, et al., “Phase III Trial Comparing Supportive Care Alone with Supportive Care with Oral Topotecan in Patients with Relapsed Small-Cell Lung Cancer,” Journal of Clinical Oncology, Vol. 24, No. 34, 2006, pp. 5441-5447. doi:10.1200/JCO.2006.06.5821
[17] D. Hartwell, J. Jones, E. Loveman, P. Harris, A. Clegg and A. Bird, “Topotecan for Relapsed Small Cell Lung Cancer: A Systematic Review and Economic Evaluation,” Cancer Treatment Reviews, Vol. 37, No. 3, 2011, pp. 242-249. doi:10.1016/j.ctrv.2010.07.005
[18] Z. A. Rasheed and E. H. Rubin, “Mechanisms of Resistance to Topoisomerase I-Targeting Drugs,” Oncogene, Vol. 22, No. 47, 2003, pp. 7296-7304.
[19] K. Fukuoka, K. Nishio, H. Fukumoto, et al., “p16INK4 Expression Is Associated with the Increased Sensitivity of Human Non-Small Cell Lung Cancer Cells to DNA Topoisomerase I Inhibitors,” Japanese Journal of Cancer Research, Vol. 88, No. 10, 1997, pp. 1009-1016. doi:10.1111/j.1349-7006.1997.tb00322.x
[20] L. Esposito, P. Indovina, F. Magnotti, D. Conti and A. Giorda, “Anticancer Therapeutic Strategies Based on CDK Inhibitors,” Current Pharmaceutical Design, 2013, in press. doi:10. 2174/ 13816128113199990377
[21] B. Schutte, L. Nieland, M. van Engeland, M. E. Henfling, L. Meijer and F. C. Ramaekers, “The Effect of the Cyclin-Dependent Kinase Inhibitor Olomoucine on Cell Cycle Kinetics,” Experimental Cell Research, Vol. 236, No. 1, 1997, pp. 4-15. doi:10.1006/excr.1997.3700
[22] D. Hanahan and R. A. Weinberg, “Hallmarks of Cancer: The Next Generation,” Cell, Vol. 144, No. 5, 2011, pp. 646-674. doi:10.1016/j.cell.2011.02.013
[23] K. X. Shu, B. Li and L. X. Wu, “The p53 Network: p53 and Its Downstream Genes,” Colloids and Surfaces B: Biointerfaces, Vol. 55, No. 1, 2007, pp. 10-18. doi:10.1016/j.colsurfb.2006.11.003
[24] R. H. Medema, R. E. Herrera, F. Lam and R. A. Weinberg, “Growth Suppression by p16ink4 Requires Functional Retinoblastoma Protein,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 92, No. 14, 1995, pp. 6289-6293. doi:10.1073/pnas.92.14.6289
[25] G. A. Otterson, R. A. Kratzke, A. Coxon, Y. W. Kim and F. J. Kaye, “Absence of p16INK4 Protein Is Restricted to the Subset of Lung Cancer Lines that Retains Wildtype RB,” Oncogene, Vol. 9, No. 11, 1994, pp. 3375-3378.
[26] H. Wikman and E. Kettunen, “Regulation of the G1/S Phase of the Cell Cycle and Alterations in the RB Pathway in Human Lung Cancer,” Expert Reviews in Anticancer Therapy, Vol. 6, No. 4, 2006, pp. 515-530. doi:10.1586/14737140.6.4.515
[27] A. Okamoto, S. P. Hussain, K. Hagiwara, et al., “Mutations in the p16INK4/MTS1/CDKN2, p15INK4B/MTS2, and p18 Genes in Primary and Metastatic Lung Cancer,” Cancer Research, Vol. 55, No. 7, 1995, pp. 1448-1451.
[28] R. Zandi, G. Selivanova and C. L. Christensen, “PRIMA1Met/APR-246 Induces Apoptosis and Tumor Growth Delay in Small Cell Lung Cancer Expressing Mutant p53,” Clinical Cancer Research, Vol. 17, No. 9, 2011, pp. 2830-2841. doi:10.1158/1078-0432.CCR-10-3168
[29] M. Abal, R. Bras-Goncalves, J. G. Judde, et al., “Enhanced Sensitivity to Irinotecan by Cdk1 Inhibition in the p53-Deficient HT29 Human Colon Cancer Cell Line,” Oncogene, Vol. 23, No. 9, 2004, pp. 1737-1744. doi:10.1038/sj.onc.1207299
[30] S. J. McClue, D. Blake, R. Clarke, et al., “In Vitro and in Vivo Antitumor Properties of the Cyclin Dependent Kinase Inhibitor CYC202 (R-Roscovitine),” International Journal of Cancer, Vol. 102, No. 5, 2002, pp. 463-468. doi:10.1002/ijc.10738
[31] A. Sharma and M. K. Bhat, “Enhancement of Carboplatinand Quercetin-Induced Cell Death by Roscovitine Is Akt Dependent and p53 Independent in Hepatoma Cells,” Integrative Cancer Therapy, Vol. 10, No. 4, 2011, pp. 4-14. doi:10.1177/1534735411423922
[32] J. W. Arends, “Molecular Interactions in the Vogelstein Model of Colorectal Carcinoma,” Journal of Pathology, Vol. 190, No. 4, 2000, pp. 412-416. doi:10.1002/(SICI)1096-9896(200003) 190:4<412::AID-PATH533>3.0.CO;2-P
[33] R. W. Johnstone, A. A. Ruefli and S. W. Lowe, “Apoptosis: A Link between Cancer Genetics and Chemotherapy,” Cell, Vol. 108, No. 2, 2002, pp. 153-164. doi:10.1016/S0092-8674(02)00625-6
[34] A. T. Sané and R. Bertrand, “Caspase Inhibition in Camptothecin-Treated U-937 Cells Is Coupled with a Shift from Apoptosis to Transient G1 Arrest Followed by Necrotic Cell Death,” Cancer Research, Vol. 59, No. 15, 1999, pp. 3565-3569.
[35] M. Motwani, C. Jung, F. M. Sirotnak, et al., “Augmentation of Apoptosis and Tumor Regression by Flavopiridol in the Presence of CPT-11 in Hct116 Colon Cancer Monolayers and Xenografts,” Clinical Cancer Research, Vol. 7, No. 12, 2001, pp. 4209-4219.
[36] K. C. Bible and S. M. Kaufmann, “Flavopiridol: A Cytotoxic Flavone That Induces Cell Death in Noncycling A549 Human Lung Carcinoma Cells,” Cancer Research, Vol. 56, No. 21, 1996, pp. 4856-4861.
[37] M. Motwani, T. M. Delohery and G. K. Schwartz, “Sequential Dependent Enhancement of Caspase Activation and Apoptosis by Flavopiridol on Paclitaxel-Treated Human Gastric and Breast Cancer Cells,” Clinical Cancer Research, Vol. 5, No. 7, 1999, pp. 1876-1883.
[38] K. C. Bible and S. H. Kaufmann, “Cytotoxic Synergy between Flavopiridol (NSC 649890, L86-8275) and Various Antineoplastic Agents: The Importance of Sequence of Administration,” Cancer Research, Vol. 57, No. 16, 1997, pp. 3375-3380.
[39] R.-G. Shao, C.-X. Cao, T. Shimizu, P. M. O’Connor, K. W. Kohn and Y. Pommier, “Abrogation of an S-Phase Checkpoint and Potentiation of Camptothecin Cytotoxicity by 7-Hydroxystaurosporine (UCN-O1) in Human Cancer Cell Lines, Possibly Influenced by p53 Function,” Cancer Research, Vol. 57, No. 18, 1997, pp. 4029-4035.
[40] M. V. Blagosklonny, “Flavopiridol, an Inhibitor of Transcription: Implications, Problems and Solutions,” Cell Cycle, Vol. 3, No. 12, 2004, pp. 1537-1542. doi:10.4161/cc.3.12.1278

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