J Vet Sci.  2009 Jun;10(2):99-103. 10.4142/jvs.2009.10.2.99.

The role of Bcl-xL and nuclear factor-kappaB in the effect of taxol on the viability of dendritic cells

• Kim MH
• Joo HG

• KMID: 1102965
• DOI: http://doi.org/10.4142/jvs.2009.10.2.99

Abstract

Taxol has been used effectively in cancer therapies. Our previous study demonstrated that taxol induced altered maturation and improved viability of dendritic cells (DCs). However, the effects of taxol on DC viability have not been fully elucidated. In the present study, flow cytometric analyses revealed that taxol treatment significantly increased the number of viable DCs and the expression levels of a representative anti-apoptotic protein Bcl-xL. Furthermore, mobilization of the p65 subunit of nuclear factor-kappaB (NF-kappaB) from the cytosol to the nucleus in DCs was observed by confocal microscopy. An inhibition assay using N-p-tosyl-L-phenylalanine chloromethyl ketone confirmed that NF-kappaB was intimately involved in the effects of taxol on DC viability. In addition, we investigated the mechanisms of taxol enhancement of DC viability. Since taxol is a popular anticancer agent used in clinic, this study may provide a rationale for the use of taxol in DC immunotherapy to treat cancer patients. Taken together, these results confirm that taxol increases DC viability, and this information may provide new insights for new clinical applications of both taxol and DCs.

Keyword

apoptosis; dendritic cell; NF-kappaB; taxol; viability

MeSH Terms

Animals
Antineoplastic Agents, Phytogenic/pharmacology
Blotting, Western
Cell Survival/drug effects/physiology
Dendritic Cells/cytology/*drug effects/physiology
Female
Flow Cytometry
Interleukin-12/physiology
Mice
Mice, Inbred BALB C
Mice, Inbred C57BL
Microscopy, Confocal
Paclitaxel/*pharmacology
Tosylphenylalanyl Chloromethyl Ketone/pharmacology
Transcription Factor RelA/antagonists & inhibitors/*physiology
Tumor Necrosis Factor-alpha/physiology
bcl-X Protein/*physiology

Figure

• 

  Fig. 1 Dendritic cell (DC) viability is enhanced by taxol treatment. DCs were cultured in the presence of medium alone or 5 µg/ml taxol for 3 days, and viable DCs were gated by cell-sized based flow cytometric analysis. The representative dot plots are presented (A), and the percentage of region R1 indicates viable DCs. Statistical analysis was performed using the data from four independent experiments (B). Asterisk (*) indicates p < 0.05 in the comparison of the medium alone-treated DCs (ContDCs) vs taxol-treated DCs (TaxolDCs).

• 

  Fig. 2 Increased cytokine production in TaxolDCs. The supernatants of DCs were harvested from culture and used for enzyme-linked immunosorbent assay. Representative data from three independent experiments are presented. Asterisk (***) indicates p < 0.001 in the comparison of medium alone vs taxol.

• 

  Fig. 3 Bcl-xL expression in dendritic cells increased after taxol treatment. Representative data from three independent experiments are presented (A). The optical density of each band was divided by that of the β-actin band, and the ratio at 0 h was set at 100% (B).

• 

  Fig. 4 NF-κB involvement in the taxol-induced effects on dendritic cells (DCs). The mobilization of NF-κB p65 molecules in DCs was detected by staining with an anti-NF-κB p65 antibody and confocal microscopy. Arrows indicate the nuclei of DCs (A). In the inhibitor assay, the percentage of viable DCs was measured as described in Fig. 1 (B). Asterisk (*) and sharp (#) indicate p < 0.05 in the comparison of ContDCs vs TaxolDCs, TaxolDCs vs TaxolDCs + N-p-tosyl-L-phenylalanine chloromethyl ketone (TPCK), respectively. Representative data from three independent experiments are presented.

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