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Antisense bcl-2 transfection up-regulates anti-apoptotic and anti-oxidant thioredoxin in neuroblastoma cells

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Abstract

Antisense bcl-2 therapy combined with chemotherapy has been proved to be effective in various tumors. However, the role played by antisense bcl-2 therapy alone is not clear. In this study, we compared the apoptosis and the protein profiles of antisense bcl-2 transfected human neuroblastoma SK-N-MC cells to the control cells. Flow cytometric data indicated that antisense bcl-2 transfection did not lead to more extensive apoptosis in SK-N-MC cells (14.9 ± 3.8%) than the control cells (10.3 ± 2.3%). The above observation was confirmed by fluorescence microscopy using Hoechst 33258 staining. However, antisense bcl-2 induced changes in the expression of various proteins as shown by proteomic comparison, which included the up-regulation of the anti-apoptotic and anti-oxidant protein thioredoxin. By western blot validation, thioredoxin was found to be up-regulated by 2.9-folds with the corresponding down-regulation of Bcl-2 by 2.1-folds. The up-regulation of thioredoxin may be a compensating mechanism for cell survival in neuroblastoma when Bcl-2 expression is suppressed, and it may to some extent attenuate the effectiveness of antisense bcl-2 therapy.

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References

  1. JM Maris KK Matthay (1999) ArticleTitleMolecular biology of neuroblastoma J Clin Oncol 17 2264–2279

    Google Scholar 

  2. GP Tonini (1993) ArticleTitleNeuroblastoma: the result of multistep transformation? Stem Cells 11 276–282

    Google Scholar 

  3. J Pritchard JA Hickman (1994) ArticleTitleWhy does stage 4s neuroblastoma regress spontaneously? Lancet 344 869–870

    Google Scholar 

  4. M Hanada S Krajewski S Tanaka D Cazals-Hatem BA Spengler RA Ross (1993) ArticleTitleRegulation of Bcl-2 oncoprotein levels with differentiation of human neuroblastoma cells Cancer Res 53 4978–4986

    Google Scholar 

  5. JC Reed L Meister S Tanaka (1991) ArticleTitleDifferential expression of Bcl 2 protooncogene in neuroblastoma and other human tumor cell lines of neural origin Cancer Res 51 6529–6538

    Google Scholar 

  6. JC Hoehner F Hedborg HJ Wiklund L Olsen S Pahlman (1995) ArticleTitleCellular death in neuroblastoma: in situ correlation of apoptosis and bcl-2 expression Int J Cancer 62 19–24

    Google Scholar 

  7. H Ikeda J Hirato M Akami S Matsuyama N Suzuki A Takahashi M Kuroiwa (1995) ArticleTitleBcl-2 oncoprotein expression and apoptosis in neuroblastoma J Pediatr Surg 30 805–808

    Google Scholar 

  8. VP Castle KP Heidelberger J Bromberg X Ou M Dole G Nunez (1993) ArticleTitleExpression of the apoptosis-suppressing protein bcl-2, in neuroblastoma is associated with unfavorable histology and N-myc amplification Am J Pathol 143 1543–1550

    Google Scholar 

  9. M Dole G Nunez AK Merchant J Maybaum CK Rode CA Bloch VP Castle (1994) ArticleTitleBcl-2 inhibits chemotherapy-induced apoptosis in neuroblastoma Cancer Res 54 3253–3259

    Google Scholar 

  10. R Jasty C Golen Particlevan HJ Lin G Solomon K Heidelberger P Polverini A Opipari E Feldman VP Castle (2001) ArticleTitleBcl-2 and M-Myc coexpression increases IGF-IR and features of malignant growth in neuroblastoma cell lines Neoplasia 3 304–313

    Google Scholar 

  11. JC Hoehner C Gestblom L Olsen S Pahlman (1997) ArticleTitleSpatial association of apoptosis-related gene expression and cellular death in clinical neuroblastoma Br J Cancer 75 1185–1194

    Google Scholar 

  12. S Krajewski J Chatten M Hanada JC Reed (1995) ArticleTitleImmunohistochemical analysis of the Bcl-2 oncoprotein in human neuroblastomas. Comparisons with tumor cell differentiation and N-Myc protein Lab Invest 72 42–54

    Google Scholar 

  13. SA Park HJ Park BI Lee YH Ahn SU Kim KS Choi (2001) ArticleTitleBcl-2 blocks cisplatin-induced apoptosis by suppression of ERK-mediated p53 accumulation in B104 cells Brain Res Mol Brain Res 93 18–26

    Google Scholar 

  14. A Lasorella A Iavarone MA Israel (1995) ArticleTitleDifferentiation of neuroblastoma enhances Bcl-2 expression and induces alterations of apoptosis and drug resistance Cancer Res 55 4711–4716

    Google Scholar 

  15. S Fulda KM Debatin (2000) ArticleTitleBetulinic acid induces apoptosis through a direct effect on mitochondria in neuroectodermal tumors Med Pediatr Oncol 35 616–668

    Google Scholar 

  16. S Fulda I Jeremias T Pietsch KM Debatin (1999) ArticleTitleBetulinic acid: a new chemotherapeutic agent in the treatment of neuroectodermal tumors Klin Padiatr 211 319–322

    Google Scholar 

  17. CJ Zhu YB Li MC Wong (2003) ArticleTitleExpression of antisense bcl-2 cDNA abolishes tumorigenicity and enhances chemosensitivity of human malignant glioma cells J Neurosci Res 74 60–66

    Google Scholar 

  18. Y Zhang XY Lei (2003) ArticleTitleEffect of bcl-2 antisense oligodeoxynucleotides on drug sensitivity of leukemic cells Hematol J 4 187–197

    Google Scholar 

  19. K Tanabe R Kim H Inoue M Emi Y Uchida T Toge (2003) ArticleTitleAntisense Bcl-2 and HER-2 oligonucleotide treatment of breast cancer cells enhances their sensitivity to anticancer drugs Int J Oncol 22 875–881

    Google Scholar 

  20. J Guan J Chen YF Luo J Gao (2002) ArticleTitleEffects of antisense bcl-2 or survivin on the growth of human neuroblastoma cell line SK-N-MC Zhonghua Yi Xue Za Zhi 82 1536–1540

    Google Scholar 

  21. J Guan J Chen H Zhao (2002) ArticleTitleEffects of tetracycline-controlled antisense bcl-2 expression on the growth and apoptosis of human neuroblastoma cell line SK-N-MC Zhonghua Bing Li Xue Za Zhi 31 135–139

    Google Scholar 

  22. S Derenne B Monia NM Dean JK Taylor MJ Rapp JL Harousseau R Bataille M Amiot (2002) ArticleTitleAntisense strategy shows that Mcl-1 rather than Bcl-2 or Bcl-x(L) is an essential survival protein of human myeloma cells Blood 100 194–199

    Google Scholar 

  23. AC Buck C Shen H Schirrmeister A Schmid-Kotsas G Munzert A Guhlmann G Mehrke N Klug HJ Gross M Bachem SN Reske (2002) ArticleTitleLiposomal delivery of antisense oligonucleotides for efficient downregulation of Bcl-2 and induction of apoptosis Cancer Biother Radiopharm 17 281–289

    Google Scholar 

  24. A Görg W Postel S Gunther (1988) ArticleTitleThe current state of two-dimensional electrophoresis with immobilized pH gradients Electrophoresis 9 531–546

    Google Scholar 

  25. M Lee DH Hyun KA Marshall LM Ellerby DE Bredesen P Jenner B Halliwell (2001) ArticleTitleEffect of overexpression of BCL-2 on cellular oxidative damage, nitric oxide production, antioxidant defenses, and the proteasome Free Radic Biol Med 31 1550–1559

    Google Scholar 

  26. SG Hwang HC Lee DW Lee YS Kim WH Joo YK Cho JY Moon (2001) ArticleTitleInduction of apoptotic cell death by a p53-independent pathway in neuronal SK-N-MC cells after treatment with 2,2′,5,5′-tetrachlorobiphenyl Toxicology 165 179–188

    Google Scholar 

  27. SH Ramirez JF Sanchez CA Dimitri HA Gelbard S Dewhurst SB Maggirwar (2001) ArticleTitleNeurotrophins prevent HIV Tat-induced neuronal apoptosis via a nuclear factor-kappaB (NF-kappaB)-dependent mechanism J Neurochem 78 874–889

    Google Scholar 

  28. AK Talley S Dewhurst SW Perry SC Dollard S Gummuluru SM Fine D New LG Epstein HE Gendelman HA Gelbard (1995) ArticleTitleTumor necrosis factor alpha-induced apoptosis in human neuronal cells: protection by the antioxidant N-acetylcysteine and the genes bcl-2 and crmA Mol Cell Biol 15 2359–2366

    Google Scholar 

  29. A Holmgren (1985) ArticleTitleThioredoxin Annu Rev Biochem 54 237–271 Occurrence Handle10.1146/annurev.bi.54.070185.001321

    Article  Google Scholar 

  30. A Holmgren (1989) ArticleTitleThioredoxin and glutaredoxin systems J Biol Chem 264 13963–13966

    Google Scholar 

  31. H Nakamura K Nakamura J Yodoi (1997) ArticleTitleRedox-regulation of cellular activation Annu Rev Immunol 15 351–369

    Google Scholar 

  32. M Hotta F Tashiro H Ikegami H Niwa T Ogiwara J Yodoi J Miyazaki (1998) ArticleTitlePancreatic β-cell-specific expression of thioredoxin, an antioxidative and antiapoptotic protein, prevents autoimmune and streptozotocin-induced diabetes J Exp Med 188 1445–1451

    Google Scholar 

  33. Y Takagi A Mitsui A Nishiyama K Nozaki H Sono Y Gon N Hashimoto J Yodoi (1999) ArticleTitleOverexpression of thioredoxin intransgenic mice attenuates focal ischemic brain damage Proc Natl Acad Sci USA 96 4131–4136

    Google Scholar 

  34. A Baker CM Payne MM Briehl G Powis (1997) ArticleTitleThioredoxin, a gene found overexpressed in human cancer, inhibits apoptosis in vitro and in vivo Cancer Res 57 5162–5167

    Google Scholar 

  35. G Powis D Mustacich A Coon (2000) ArticleTitleThe role of the redox protein thioredoxin in cell growth and cancer Free Radic Biol Med. 29 312–322

    Google Scholar 

  36. M Saitoh H Nishitoh M Fujii K Takeda K Tobiume Y Sawada (1998) ArticleTitleMammalian thioredoxin is a direct inhibitor of apoptosis signal-regulating kinase (ASK) 1 EMBO J 17 2596–2606 Occurrence Handle10.1093/emboj/17.9.2596

    Article  Google Scholar 

  37. K Tobiume A Matsuzawa T Takahashi H Nishitoh K Morita K Takeda O Minowa K Miyazono T Noda H Ichijo (2001) ArticleTitleASK1 is required for sustained activations of JNK/p38 MAP kinases and apoptosis EMBO Rep 2 222–228

    Google Scholar 

  38. H Matsuura H Nishitoh K Takeda A Matsuzawa T Amagasa M Ito K Yoshioka H Ichijo (2002) ArticleTitlePhosphorylation-dependent scaffolding role of JSAP1/JIP3 in the ASK1-JNK signaling pathway. A new mode of regulation of the MAP kinase cascade J Biol Chem 277 40703–40709

    Google Scholar 

  39. NJ Hodges D Smart AJ Lee NA Lewis JK Chipman (2004) ArticleTitleActivation of c-Jun N-terminal kinase in A549 lung carcinoma cells by sodium dichromate: role of dissociation of apoptosis signal regulating kinase-1 from its physiological inhibitor thioredoxin Toxicology 197 101–112

    Google Scholar 

  40. YJ Lee JH Kim J Chen JJ Song (2002) ArticleTitleEnhancement of metabolic oxidative stress-induced cytotoxicity by the thioredoxin inhibitor 1-methylpropyl 2-imidazolyl disulfide is mediated through the ASK1-SEK1-JNK1 pathway Mol Pharmacol 62 1409–1417

    Google Scholar 

  41. CR Weston RJ Davis (2002) ArticleTitleThe JNK signal transduction pathway Curr Opin Genet Dev 12 14–21

    Google Scholar 

  42. M Fan M Goodwin T Vu C Brantley-Finley WA Gaarde TC Chambers (2000) ArticleTitleVinblastine-induced phosphorylation of Bcl-2 and Bcl-XL is mediated by JNK and occurs in parallel with inactivation of the Raf-1/MEK/ERK cascade J Biol Chem 275 29980–29985

    Google Scholar 

  43. T Andoh CC Chiueh PB Chock (2003) ArticleTitleCyclic GMP-dependent protein kinase regulates the expression of thioredoxin and thioredoxin peroxidase-1 during hormesis in response to oxidative stress-induced apoptosis J Biol Chem 278 885–890

    Google Scholar 

  44. S Kikuchi K Shinpo M Niino S Tsuji K Iwabuchi K Onoe K Tashiro (2002) ArticleTitleProstaglandin E1 protects cultured spinal neurons against the effects of nitric oxide toxicity Neuropharmacology 42 714–723

    Google Scholar 

  45. Filomeni G, Rotilio G, Ciriolo MR: FASEB J: Glutathione disulfide induces apoptosis in U937 cells by a redox-mediated p38 MAP kinase pathway. 17: 64–66, 2003

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Authors and Affiliations

  1. Department of Pathology, Peking Union Medical College Hospital, Beijing, P.R. China

    Yiting Li, Zhaohui Lu, Fang Chen, Jian Guan, Yanfeng Xu & Jie Chen

  2. National Laboratory of Medical Molecular Biology, Institute of Basic Medicine, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, P.R. China

    Liping Hu

  3. Texas Children’s Hospital, 6621 Fannin Street, MC 3-3320, Houston, TX, 77030, USA

    Yiting Li

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  1. Yiting Li
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  2. Zhaohui Lu
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Correspondence to Yiting Li.

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Li, Y., Lu, Z., Chen, F. et al. Antisense bcl-2 transfection up-regulates anti-apoptotic and anti-oxidant thioredoxin in neuroblastoma cells. J Neurooncol 72, 17–23 (2005). https://doi.org/10.1007/s11060-004-3116-x

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