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Mol Med. 1999 November; 5(11): 711–720.
PMCID: PMC2230485

beta-lapachone induces cell cycle arrest and apoptosis in human colon cancer cells.

Abstract

BACKGROUND: Human colon cancers have a high frequency of p53 mutations, and cancer cells expressing mutant p53 tend to be resistant to current chemo- and radiation therapy. It is thus important to find therapeutic agents that can inhibit colon cancer cells with altered p53 status. beta-Lapachone, a novel topoisomerase inhibitor, has been shown to induce cell death in human promyelocytic leukemia and prostate cancer cells through a p53-independent pathway. Here we examined the effects of beta-lapachone on human colon cancer cells. MATERIALS AND METHODS: Several human colon cancer cell lines, SW480, SW620, and DLD1, with mutant or defective p53, were used. The antiproliferative effects of beta-lapachone were assessed by colony formation assays, cell cycle analysis, and apoptosis analysis, including annexin V staining and DNA laddering analysis. The effects on cell cycle and apoptosis regulatory proteins were examined by immunoblotting. RESULTS: All three cell lines, SW480, SW620, and DLD1, were sensitive to beta-lapachone, with an IC(50) of 2 to 3 microM in colony formation assays, a finding similar to that previously reported for prostate cancer cells. However, these cells were arrested in different stages of S phase. At 24 hr post-treatment, beta-lapachone induced S-, late S/G2-, and early S-phase arrest in SW480, SW620, and DLD1 cells, respectively. The cell cycle alterations induced by beta-lapachone were congruous with changes in cell cycle regulatory proteins such as cyclin A, cyclin B1, cdc2, and cyclin D1. Moreover, beta-lapachone induced apoptosis, as demonstrated by annexin V staining, flow cytometric analysis of DNA content, and DNA laddering analysis. Furthermore, down-regulation of mutant p53 and induction of p27 in SW480 cells, and induction of pro-apoptotic protein Bax in DLD1 cells may be pertinent to the anti-proliferative and apoptotic effects of beta-lapachone on these cells. CONCLUSIONS: beta-Lapachone induced cell cycle arrest and apoptosis in human colon cancer cells through a p53-independent pathway. For human colon cancers, which often contain p53 mutations, beta-lapachone may prove to be a promising anticancer agent that can target cancer cells, especially those with mutant p53.

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Selected References

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  • Cruz FS, Docampo R, Boveris A. Generation of superoxide anions and hydrogen peroxide from beta-lapachone in bacteria. Antimicrob Agents Chemother. 1978 Oct;14(4):630–633. [PMC free article] [PubMed]
  • Guiraud P, Steiman R, Campos-Takaki GM, Seigle-Murandi F, Simeon de Buochberg M. Comparison of antibacterial and antifungal activities of lapachol and beta-lapachone. Planta Med. 1994 Aug;60(4):373–374. [PubMed]
  • Schuerch AR, Wehrli W. beta-Lapachone, an inhibitor of oncornavirus reverse transcriptase and eukaryotic DNA polymerase-alpha. Inhibitory effect, thiol dependence and specificity. Eur J Biochem. 1978 Mar;84(1):197–205. [PubMed]
  • Li CJ, Zhang LJ, Dezube BJ, Crumpacker CS, Pardee AB. Three inhibitors of type 1 human immunodeficiency virus long terminal repeat-directed gene expression and virus replication. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1839–1842. [PubMed]
  • Planchon SM, Wuerzberger S, Frydman B, Witiak DT, Hutson P, Church DR, Wilding G, Boothman DA. Beta-lapachone-mediated apoptosis in human promyelocytic leukemia (HL-60) and human prostate cancer cells: a p53-independent response. Cancer Res. 1995 Sep 1;55(17):3706–3711. [PubMed]
  • Li CJ, Wang C, Pardee AB. Induction of apoptosis by beta-lapachone in human prostate cancer cells. Cancer Res. 1995 Sep 1;55(17):3712–3715. [PubMed]
  • Weller M, Winter S, Schmidt C, Esser P, Fontana A, Dichgans J, Groscurth P. Topoisomerase-I inhibitors for human malignant glioma: differential modulation of p53, p21, bax and bcl-2 expression and of CD95-mediated apoptosis by camptothecin and beta-lapachone. Int J Cancer. 1997 Nov 27;73(5):707–714. [PubMed]
  • Lai CC, Liu TJ, Ho LK, Don MJ, Chau YP. beta-Lapachone induced cell death in human hepatoma (HepA2) cells. Histol Histopathol. 1998 Jan;13(1):89–97. [PubMed]
  • Wuerzberger SM, Pink JJ, Planchon SM, Byers KL, Bornmann WG, Boothman DA. Induction of apoptosis in MCF-7:WS8 breast cancer cells by beta-lapachone. Cancer Res. 1998 May 1;58(9):1876–1885. [PubMed]
  • Li YZ, Li CJ, Pinto AV, Pardee AB. Release of mitochondrial cytochrome C in both apoptosis and necrosis induced by beta-lapachone in human carcinoma cells. Mol Med. 1999 Apr;5(4):232–239. [PMC free article] [PubMed]
  • Li CJ, Li YZ, Pinto AV, Pardee AB. Potent inhibition of tumor survival in vivo by beta-lapachone plus taxol: combining drugs imposes different artificial checkpoints. Proc Natl Acad Sci U S A. 1999 Nov 9;96(23):13369–13374. [PubMed]
  • Li CJ, Averboukh L, Pardee AB. beta-Lapachone, a novel DNA topoisomerase I inhibitor with a mode of action different from camptothecin. J Biol Chem. 1993 Oct 25;268(30):22463–22468. [PubMed]
  • Hsiang YH, Hertzberg R, Hecht S, Liu LF. Camptothecin induces protein-linked DNA breaks via mammalian DNA topoisomerase I. J Biol Chem. 1985 Nov 25;260(27):14873–14878. [PubMed]
  • Frydman B, Marton LJ, Sun JS, Neder K, Witiak DT, Liu AA, Wang HM, Mao Y, Wu HY, Sanders MM, et al. Induction of DNA topoisomerase II-mediated DNA cleavage by beta-lapachone and related naphthoquinones. Cancer Res. 1997 Feb 15;57(4):620–627. [PubMed]
  • Wang JC. DNA topoisomerases. Annu Rev Biochem. 1996;65:635–692. [PubMed]
  • Pommier Y. Diversity of DNA topoisomerases I and inhibitors. Biochimie. 1998 Mar;80(3):255–270. [PubMed]
  • Boorstein RJ, Pardee AB. Beta-lapachone greatly enhances MMS lethality to human fibroblasts. Biochem Biophys Res Commun. 1984 Feb 14;118(3):828–834. [PubMed]
  • Boothman DA, Greer S, Pardee AB. Potentiation of halogenated pyrimidine radiosensitizers in human carcinoma cells by beta-lapachone (3,4-dihydro-2,2-dimethyl-2H-naphtho[1,2-b]pyran- 5,6-dione), a novel DNA repair inhibitor. Cancer Res. 1987 Oct 15;47(20):5361–5366. [PubMed]
  • Boothman DA, Trask DK, Pardee AB. Inhibition of potentially lethal DNA damage repair in human tumor cells by beta-lapachone, an activator of topoisomerase I. Cancer Res. 1989 Feb 1;49(3):605–612. [PubMed]
  • Degrassi F, De Salvia R, Berghella L. The production of chromosomal alterations by beta-lapachone, an activator of topoisomerase I. Mutat Res. 1993 Aug;288(2):263–267. [PubMed]
  • Lowe SW, Bodis S, McClatchey A, Remington L, Ruley HE, Fisher DE, Housman DE, Jacks T. p53 status and the efficacy of cancer therapy in vivo. Science. 1994 Nov 4;266(5186):807–810. [PubMed]
  • Lowe SW. Cancer therapy and p53. Curr Opin Oncol. 1995 Nov;7(6):547–553. [PubMed]
  • O'Connor PM, Jackman J, Bae I, Myers TG, Fan S, Mutoh M, Scudiero DA, Monks A, Sausville EA, Weinstein JN, et al. Characterization of the p53 tumor suppressor pathway in cell lines of the National Cancer Institute anticancer drug screen and correlations with the growth-inhibitory potency of 123 anticancer agents. Cancer Res. 1997 Oct 1;57(19):4285–4300. [PubMed]
  • Cho KR, Vogelstein B. Suppressor gene alterations in the colorectal adenoma-carcinoma sequence. J Cell Biochem Suppl. 1992;16G:137–141. [PubMed]
  • Sinicrope FA, Roddey G, Lemoine M, Ruan S, Stephens LC, Frazier ML, Shen Y, Zhang W. Loss of p21WAF1/Cip1 protein expression accompanies progression of sporadic colorectal neoplasms but not hereditary nonpolyposis colorectal cancers. Clin Cancer Res. 1998 May;4(5):1251–1261. [PubMed]
  • Sheikh MS, Chen YQ, Smith ML, Fornace AJ., Jr Role of p21Waf1/Cip1/Sdi1 in cell death and DNA repair as studied using a tetracycline-inducible system in p53-deficient cells. Oncogene. 1997 Apr 17;14(15):1875–1882. [PubMed]
  • Dignam JD, Lebovitz RM, Roeder RG. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. [PMC free article] [PubMed]
  • Martin SJ, Reutelingsperger CP, McGahon AJ, Rader JA, van Schie RC, LaFace DM, Green DR. Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J Exp Med. 1995 Nov 1;182(5):1545–1556. [PMC free article] [PubMed]
  • Pagano M, Theodoras AM, Tam SW, Draetta GF. Cyclin D1-mediated inhibition of repair and replicative DNA synthesis in human fibroblasts. Genes Dev. 1994 Jul 15;8(14):1627–1639. [PubMed]
  • Bates S, Vousden KH. p53 in signaling checkpoint arrest or apoptosis. Curr Opin Genet Dev. 1996 Feb;6(1):12–18. [PubMed]
  • Yang ZY, Perkins ND, Ohno T, Nabel EG, Nabel GJ. The p21 cyclin-dependent kinase inhibitor suppresses tumorigenicity in vivo. Nat Med. 1995 Oct;1(10):1052–1056. [PubMed]
  • Ahmad N, Feyes DK, Agarwal R, Mukhtar H. Photodynamic therapy results in induction of WAF1/CIP1/P21 leading to cell cycle arrest and apoptosis. Proc Natl Acad Sci U S A. 1998 Jun 9;95(12):6977–6982. [PubMed]
  • Katayose Y, Kim M, Rakkar AN, Li Z, Cowan KH, Seth P. Promoting apoptosis: a novel activity associated with the cyclin-dependent kinase inhibitor p27. Cancer Res. 1997 Dec 15;57(24):5441–5445. [PubMed]
  • Korsmeyer SJ. Regulators of cell death. Trends Genet. 1995 Mar;11(3):101–105. [PubMed]
  • Vanni A, Fiore M, De Salvia R, Cundari E, Ricordy R, Ceccarelli R, Degrassi F. DNA damage and cytotoxicity induced by beta-lapachone: relation to poly(ADP-ribose) polymerase inhibition. Mutat Res. 1998 Jun 5;401(1-2):55–63. [PubMed]
  • Dittmer D, Pati S, Zambetti G, Chu S, Teresky AK, Moore M, Finlay C, Levine AJ. Gain of function mutations in p53. Nat Genet. 1993 May;4(1):42–46. [PubMed]
  • Liu PK, Kraus E, Wu TA, Strong LC, Tainsky MA. Analysis of genomic instability in Li-Fraumeni fibroblasts with germline p53 mutations. Oncogene. 1996 Jun 6;12(11):2267–2278. [PMC free article] [PubMed]

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