Home Topotecan and methotrexate alter expression of the apoptosis-related genes BCL2, FAS and BCL2L12 in leukemic HL-60 cells
Article
Licensed
Unlicensed Requires Authentication

Topotecan and methotrexate alter expression of the apoptosis-related genes BCL2, FAS and BCL2L12 in leukemic HL-60 cells

  • Kostas V. Floros , Maroulio Talieri and Andreas Scorilas
Published/Copyright: November 28, 2006
Biological Chemistry
From the journal Volume 387 Issue 12

Abstract

The BCL2 family of genes (B-cell CLL/lymphoma 2; Bcl-2) plays a pivotal role in the highly regulated process of apoptosis. We have recently cloned a newly identified member of this family, BCL2L12, which was found to be differentially expressed in many tumors. It is known that topotecan and methotrexate act through induction of apoptosis in cancer cells. In the present study we investigated the expression profile of the novel apoptotic gene BCL2L12 in relation to other apoptotic genes in the human leukemic cell line HL-60, after treatment with topotecan or methotrexate. The kinetics of apoptosis induction and cell toxicity were investigated by DNA laddering and the MTT method, respectively. Gene expression levels were analyzed by RT-PCR using gene-specific primers. Downregulation of BCL2L12, BCL2 and FAS was observed after treatment of HL-60 cells with topotecan, while treatment with methotrexate led to downregulation of BCL2 and FAS, with no change in BCL2L12 expression. Our results support the significance of mRNA modulations in the expression of apoptosis-related genes during treatment of human leukemic cells with anticancer drugs.

:

Corresponding author ;

References

Baliga, B.C. and Kumar, S. (2002). Role of BCL-2 family of proteins in malignancy. Hematol. Oncol.20, 63–74.10.1002/hon.685Search in Google Scholar

Barry, M.A., Behnke, C.A., and Eastman, A. (1990). Activation of programmed cell death (apoptosis) by cisplatin, other anticancer drugs, toxins and hyperthermia. Biochem. Pharmacol.40, 2353–2362.10.1016/0006-2952(90)90733-2Search in Google Scholar

Beran, M., Estey, E., O'Brien, S., Cortes, J., Koller, C.A., Giles, F.J., Kornblau, S., Andreeff, M., Vey, N., Pierce, S.R., et al. (1999). Topotecan and cytarabine is an active combination regimen in myelodysplastic syndromes and chronic myelomonocytic leukemia. J. Clin. Oncol.17, 2819–2830.10.1200/JCO.1999.17.9.2819Search in Google Scholar

Cheng, E.H.Y., Levine, B., Boise, L.H., Thompson, C.B., and Hardwick, J.M. (1996). Bax-independent inhibition of apoptosis by Bcl-XL. Nature379, 554–556.10.1038/379554a0Search in Google Scholar

Chomczynski, P. and Sacchi, N. (1987). Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem.162, 156–159.10.1016/0003-2697(87)90021-2Search in Google Scholar

de Silva, C.P., de Oliveira, C.R., and de Conceicao, P.L. (1996). Apoptosis as a mechanism of cell death induced by different chemotherapeutic drugs in human leukemic T-lymphocytes. Biochem. Pharmacol.51, 1331–1340.10.1016/0006-2952(96)00041-XSearch in Google Scholar

Farber, S., Toch, R., Sears, E., and Pinkel, D. (1956). Advances in chemotherapy of cancer in man. In: Advances in Cancer Research, H.A. Greenstein, ed. (New York, USA: Academic Press), pp. 2–73.10.1016/S0065-230X(08)60721-6Search in Google Scholar

Floros, V.K., Thomadaki, H., Katsaros, N., Talieri, M., and Scorilas, A. (2004). mRNA expression analysis of a variety of apoptosis-related genes, including the novel gene of the BCL2 family, BCL2L12, in HL-60 leukemia cells after treatment with carboplatin and doxorubicin. Biol. Chem.385, 1099–1103.10.1515/BC.2004.143Search in Google Scholar PubMed

Floros, V.K., Thomadaki, H., Lallas, G., Katsaros, N., Talieri, M., and Scorilas, A. (2003). Cisplatin-induced apoptosis in HL-60 human promyelocytic leukemia cells. Differential expression of BCL2 and novel apoptosis-related gene BCL2L12. Ann. NY Acad. Sci.1010, 153–158.10.1196/annals.1299.025Search in Google Scholar PubMed

Friesen, C., Herr, I., Krammer, P.H., and Debatin, K.-M. (1996). Involvement of the CD95 APO-1/Fas receptor/ligand system in drug-induced apoptosis in leukemia cells. Nat. Med.2, 574–577.10.1038/nm0596-574Search in Google Scholar PubMed

Gazzanelli, G., Luchetti, F., Burattini, S., Mannello, F., Falcieri E., and Papa, S. (2000). Matrix metalloproteinases expression in HL-60 promyelocytic leukemia cells during apoptosis. Apoptosis5, 165–172.10.1023/A:1009688831531Search in Google Scholar

Genestier, L., Paillot, R., Quemecuer, L., Izeradjene, K., and Revillard, J. (2000). Mechanisms of action of methotrexate. Immunopharmacology4, 247–257.10.1016/S0162-3109(00)00189-2Search in Google Scholar

Graham, P. and Stevenson, F. (2005). Bodyguards and assassins: Bcl-2 family proteins and apoptosis control in chronic lymphocytic leukaemia. Immunology114, 441–449.Search in Google Scholar

Harada, K., Ishibashi, S., Miyashita, T., Osuga, J., Yagyu, H., Ohashi, K., Yazaki, Y., and Yamada, N. (2002). Bcl-2 protein inhibits oxysterol-induced apoptosis through suppressing CPP32-mediated pathway. FEBS Lett.411, 63–66.Search in Google Scholar

Kalomenidis, I., Orphanidou, D., Papamichalis, G., Vassilakopoulos, T., Scorilas, A., Rasidakis, A., Papastamatiou, H., Jordanoglou, J., and Roussos, C. (2001). Combined expression of p53, Bcl-2, and p21WAF-1 proteins in lung cancer and premalignant lesions: association with clinical characteristics. Lung179, 265–278.10.1007/s004080010067Search in Google Scholar

Kasibhatla, S., Brunner, T., Genestier, L., Echeverri, F., Mahboubi, A., and Green, D.R. (1998). DNA damaging agents induce expression of Fas ligand and subsequent apoptosis in T lymphocytes via the activation of NF-κB and AP-1. Mol. Cell1, 543–551.10.1016/S1097-2765(00)80054-4Search in Google Scholar

Kaufmann, S. (1989). Induction of endonucleolytic DNA cleavage in human acute myelogenous leukemia cells by etoposide, camptothecin and other cytotoxic anticancer drugs: a cautionary note. Cancer Res.49, 5870–5878.Search in Google Scholar

Kratzmeier, M., Albig, W., Hanecke, K., and Doenecke, D. (2000). Rapid dephosphorylation of H1 histones after apoptosis induction. J. Biol. Chem.275, 30478–30486.10.1074/jbc.M003956200Search in Google Scholar PubMed

Mathioudaki, K., Scorilas, A., Papadokostopoulou, A., Xynopoulos, D., Arnogianaki, N., Agnanti, N., and Talieri, M. (2004). Expression analysis of BCL2L12, a new member of apoptosis-related genes, in colon cancer. Biol. Chem.385, 779–783.10.1515/BC.2004.101Search in Google Scholar PubMed

Mirowski, M., Rozalski, M., Krajewska, U., Balcerczak, E., Mlynarski, W., and Wierzbicki, R. (2003). Induction of caspase 3 and modulation of some apoptotic genes in human acute promyelocytic leukemia HL-60 cells by carboplatin with amifostine. Pol. J. Pharmacol.55, 227–234.Search in Google Scholar

Muller, M., Strand, S., Hug, H., Heinemann, E.M., Walczak, H., Hofmann, W.J., Stremmel, W., Krammer, P.H., and Galle, P.R., (1997). Drug-induced apoptosis in hepatoma cells is mediated by the CD95 (APO-1/Fas) receptor ligand system and involves activation of wild-type p53. J. Clin. Invest.99, 403–413.10.1172/JCI119174Search in Google Scholar PubMed PubMed Central

Muller, M., Wilder, S., Bannasch, D., Israeli, D., Lehlbach, K., Li-Weber, M., Friedman, S.L., Galle, P.R., Stremmel, W., Oren, M., and Krammer, P.H. (1998). p53 activates the CD95. J. Exp. Med.188, 2033–2045.Search in Google Scholar

Reed, J.C. (1997). Double identity for proteins of the Bcl-2-family. Nature387, 773–776.10.1038/42867Search in Google Scholar PubMed

Revelos, K., Petraki, C., Gregorakis, A., Scorilas, A., Papanastasiou, P., and Koutsilieris, M. (2005). Immunohistochemical expression of Bcl2 is an independent predictor of time-to-biochemical failure in patients with clinically localized prostate cancer following radical prostatectomy. Anticancer Res.25, 3123–3133.Search in Google Scholar

Rowinsky, E.K., Kaufmann, S.H., Baker, S.D., Miller, C.B., Sartorius, S.E., Bowling, M.K., Chen, T.L., Donehower, R.C., and Gore, S.D. (1996). A phase I and pharmacological study of topotecan infused over 30 minutes for five days in patients with refractory acute leukemia. Clin. Cancer Res.2, 1921–1930.Search in Google Scholar

Schliephacke, T., Meinl, A., Kratzmeier, M., Doenecke D., and Albig, W. (2004). The telomeric region is excluded from nucleosomal fragmentation during apoptosis, but the bulk nuclear chromatin is randomly degraded. Cell Death Differ.11, 693–703.10.1038/sj.cdd.4401414Search in Google Scholar PubMed

Scorilas, A., Kyriakopoulou, L., Yousef, G.M., Ashworth, L., Kwamie, A., and Diamandis, E.P. (2001). Molecular cloning, physical mapping and expression analysis of a novel gene, BCL2L12, encoding for a proline-rich protein with a highly conserved BH2 domain of the BCL2 family. Genomics72, 217–221.10.1006/geno.2000.6455Search in Google Scholar PubMed

Simonian, P.L., Grillot, D.A.M., Merino, R., and Nunez, G. (1996). Bax can antagonize Bcl-XL during etoposide- and cisplatin-induced cell death independently of its heterodimerization with Bcl-XL. J. Biol. Chem.271, 22764–22772.10.1074/jbc.271.37.22764Search in Google Scholar PubMed

Talieri, M., Diamandis, E.P., Katsaros, N., Gourgiotis, D., Karida, I., and Scorilas, A. (2003). Expression of BCL2L12, a new member of apoptosis-related proteins, in breast tumors. Thromb. Haemost.89, 1081–1088.10.1055/s-0037-1613411Search in Google Scholar

Thomadaki, H. and Scorilas, A. (2006). BCL2 family of apoptosis-related genes: functions and clinical implications in cancer. Crit. Rev. Clin. Lab. Sci.43, 1–67.Search in Google Scholar

Thomadaki, H., Tsiapalis, C.M., and Scorilas, A. (2005). Polyadenylate polymerase modulations in human epithelioid cervix and breast cancer cell lines, treated with etoposide or cordycepin, follow cell cycle rather than apoptosis induction. Biol. Chem.386, 471–480.10.1515/BC.2005.056Search in Google Scholar PubMed

Thomadaki, H., Talieri, M., and Scorilas, A. (2006). Prognostic value of the apoptosis related genes BCL2 and BCL2L12 in breast cancer. Cancer Lett., in press.Search in Google Scholar

Published Online: 2006-11-28
Published in Print: 2006-12-01

©2006 by Walter de Gruyter Berlin New York

Articles in the same Issue

  1. Janus-faced role of endothelial NO synthase in vascular disease: uncoupling of oxygen reduction from NO synthesis and its pharmacological reversal
  2. Leucine aminopeptidases: diversity in structure and function
  3. Endogenous anti-inflammatory substances, inter-α-inhibitor and bikunin
  4. Mitochondrial morphology and distribution in mammalian cells
  5. Heterogeneity in the cysteine protease inhibitor clitocypin gene family
  6. Mutations in the inter-SH2 domain of the regulatory subunit of phosphoinositide 3-kinase: effects on catalytic subunit binding and holoenzyme function
  7. Evaluation of Bacillus anthracis thymidine kinase as a potential target for the development of antibacterial nucleoside analogs
  8. The human malaria parasite Plasmodium falciparum expresses an atypical N-terminally extended pyrophosphokinase with specificity for thiamine
  9. Fes1p acts as a nucleotide exchange factor for the ribosome-associated molecular chaperone Ssb1p
  10. Detailed analysis of MIA protein by mutagenesis
  11. The role of human tissue kallikreins 7 and 8 in intracranial malignancies
  12. Prognostic significance of the expression of SR-A1, encoding a novel SR-related CTD-associated factor, in breast cancer
  13. Suppression of TNF-α production by S-adenosylmethionine in human mononuclear leukocytes is not mediated by polyamines
  14. Topotecan and methotrexate alter expression of the apoptosis-related genes BCL2, FAS and BCL2L12 in leukemic HL-60 cells
  15. Two secreted cystatins of the soft tick Ornithodoros moubata: differential expression pattern and inhibitory specificity
  16. Acknowledgment
  17. Contents Biological Chemistry Volume 387, 2006
  18. Author Index
  19. Subject Index
Downloaded on 10.9.2025 from https://www.degruyterbrill.com/document/doi/10.1515/BC.2006.203/html
Scroll to top button