Drug Res (Stuttg) 2013; 63(07): 362-369
DOI: 10.1055/s-0033-1341463
Original Article
© Georg Thieme Verlag KG Stuttgart · New York

Novel Quinuclidinone Derivatives Induce Apoptosis in Lung Cancer via Sphingomyelinase Pathways

A. Malki
1   Biochemistry department, Faculty of Science, Alexandria University, Alexandria, Egypt
L. Fathy
2   Chemistry department, Faculty of Science, Alexandria University, Alexandria, Egypt
E.S H. El Ashry
2   Chemistry department, Faculty of Science, Alexandria University, Alexandria, Egypt
› Author Affiliations
Further Information

Publication History

received 07 March 2012

accepted 06 March 2013

Publication Date:
12 April 2013 (online)


We previously reported novel quinuclidinone analogs which induced apoptosis in lung and breast cancer cells. In this study, we designed and synthesized novel quinuclidinone analogs that showed cytotoxicity in lung cancer cells. The effects of these analogs were studied in H1299 human large cell lung carcinoma cells that are null for p53 and normal lung epithelial cell lines (NL-20). The effects of the analogs were investigated by MTT assay, ELISA based apoptotic assay, TUNEL assay, sphingomylinase activity, flow cytometry and western blot analysis. Our data indicated that derivatives 4 and 6 decreased cell proliferation and induced apoptosis in H1299 cells more than NL-20 cells. Derivatives 4 and 6 reduced percent of cells in G2/M phase in H1299 cells more than NL-20 cells and these results were confirmed by increased expression levels of cyclin E. Furthermore, derivatives 4 and 6 increased sphingomyelinase activity, caspase-8, and caspase-9 and JNK-1 expression level in H1299. Additionally, derivatives 4 and 6 induced Procaspase-3, PARP-1 cleavage, and increased caspase-3 activity. All these results confirm that our quinuclidinone derivatives provoke cytotoxicity in lung cancer cells through the interplay of key apoptosis molecules in different compartments of the cell beginning with an increase in sphingomyelinase activity.

  • References

  • 1 Jemal A, Thomas A, Murray T et al. Cancer statistics. CA Cancer J Clin 2005; 52: 23-27
  • 2 Carney DN. Lung cancer – time to move on from chemotherapy. N Engl J Med 2002; 346: 126-128
  • 3 Thomas P, Rubinstein L. Cancer recurrence after resection. T1 N0 non-small cell lung cancer. Lung Cancer Study Group Ann Thorac Surg 1990; 49: 242-246
  • 4 Travis WD, Travis LB, Devesa SS. Lung cancer. Cancer 1995; 75: 191-192
  • 5 Dunst J. Role of radiotherapy in small cell lung cancer. Lung Cancer 2001; 33: 137-141
  • 6 Cohen V, Khuri FR. Chemoprevention of lung cancer. Curr Opin Pulm Med 2004; 10: 279-283
  • 7 Hanahan D, Weinberg RA. The hallmarks of cancer. Cell 2000; 100: 57-70
  • 8 Liscovitch M, Cantley LC. Lipid second messengers. Cell 1994; 77: 329-334
  • 9 Saddoughi SA, Song P, Ogretmen B. Roles of bioactive sphingolipids in cancer biology and therapeutics. Subcell Biochem 2008; 49: 413-440
  • 10 Smyth ML, Obeid LM, Hannun YA. Ceramide a novel lipid mediator of apoptosis. Adv Pharmacol 1997; 41: 33-54
  • 11 Sonnino S, ureli M, Loberto N et al. Fine tuning of cell functions through remodeling of glycosphingolipids by plasma membrane-associated glycohydrolases. FEBS Lett 2010; 584: 1914-1922
  • 12 Lacour S, Hammann A, Grazide S et al. Cisplatin-induced CD95 redistribution into membrane lipid rafts of HT29 human colon cancer cells. Cancer Res 2004; 64: 3593-3598
  • 13 Morita Y, Perez GI, Paris F. Oocyte apoptosis is suppressed by disruption of the acid sphingomyelinase gene or by sphingosine-1-phosphate therapy. Nat Med 2000; 6: 1109-1114
  • 14 Oliver L, Vallette FM. The role of caspases in cell death and differentiation. Drug Resist Updat 2005; 8: 163-170
  • 15 Fischer U, Janicke RU, Schulze-Osthoff K. Many cuts to ruin: a comprehensive update of caspase substrates. Cell Death Differ 2003; 10: 76-100
  • 16 Bykov VJ, Issaeva N, Selivanova G et al. Mutant p53-dependent growth suppression distinguishes PRIMA-1 from known anticancer drugs: a statistical analysis of information in the National Cancer Institute database. Carcinogenesis 2002; 12: 2011-2018
  • 17 Bykov V, Issaeva N, Shilov NA et al. Restoration of the tumor suppressor function to mutant p53 by a low-molecular-weight compound. Nat Med 2000; 8: 282-288
  • 18 Malki A, Bergmeier S. Differential Apoptotic Effects of Novel Quinuclidinone analogs 8a and 8b in Normal and Lung Cancer Cell Lines. Anticancer Res 2011; 31: 1345-1358
  • 19 Malki A, Bergmeier S. Novel Quinuclidinone Derivative 8a Induced Apoptosis in Human MCF-7 Breast Cancer Cell Lines. Anticancer Res 2011; 31: 871-881
  • 20 Nielsen AT. Systems with bridgehead nitrogen. B-Ketols of the 1-azabicyclo[2.2.2]octane series. J Org Chem 1966; 31: 1053-1059
  • 21 Bondarenko VA, Mikhlina EE, Filipenko T et al. Reaction of 2-methylene-3-oxoquinuclidine with bifunctional nucleophilic reagents. Khim Getero Soed 1979; 10: 1393-1397
  • 22 Vazquez A, Bond EE, Levine AJ et al. The genetics of the p53 pathway, apoptosis and cancer therapy. Nat Rev Drug Discov 2008; 7: 979-987
  • 23 Benner SE, Hong WK. Clinical chemoprevention: developing a cancer prevention strategy. J Natl Cancer Inst (Bethesda) 1993; 85: 1446-1447
  • 24 Elledge SJ. Cell cycle checkpoints: preventing an identity crisis. Science 1996; 274: 1664-1672
  • 25 Longley DB, Johnston PG. Molecular mechanisms of drug resistance. J Pathol 2005; 205: 275-292
  • 26 Lampson MA, Kapoor TM. Unraveling cell division mechanisms with small-molecule inhibitors. Nat Chem Biol 2006; 2: 19-27
  • 27 Yu Q, La Rose J, Zhang H et al. UCN-01 inhibits p53 up-regulation and abrogates gamma-radiation-induced G(2)-M checkpoint independently of p53 by targeting both of the checkpoint kinases, Chk2 and Chk1. Cancer Res 2002; 62: 5743-5748
  • 28 Aldridge DR, Radford IR. Explaining differences in sensitivity to killing by ionizing radiation between human lymphoid cell lines. Cancer Res 1998; 58: 2817-2824
  • 29 Gulbins E, Bissonnette R, Mahboubi A et al. FAS-induced apoptosis is mediated via a ceramide-initiated RAS signaling pathway. Immunity 1995; 2: 341-351
  • 30 Hannun YA, Obeid LM. Ceramide: an intracellular signal for apoptosis. Trends Biochem Sci 1995; 20: 73-77
  • 31 Jarvis WD, Kolesnick RN, Fornari FA et al. Induction of apoptotic DNA damage and cell death by activation of the sphingomyelin pathway. Proc Natl Acad Sci USA 1994; 91: 73-77
  • 32 Hannun YA, Linardic V. Sphingolipid breakdown products: anti-proliferative and tumor-suppressor lipids. Biochimica et Biophysica Acta 1993; 1154: 223-236
  • 33 Oliver L, Vallette FM. The role of caspases in cell death and differentiation. Drug Resist Update 2005; 8: 163-170
  • 34 Fischer U, Janicke RU, Schulze-Osthoff K. Many cuts to ruin: a comprehensive update of caspase substrates. Cell Death Differ 2003; 10: 76-100