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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (3): 249-254.doi: 10.3736/jcim20090310

• Original Experimental Research • Previous Articles     Next Articles

Effects of Feiyanning Decoction on gene expression of nuclear factor-κB activated by tumor necrosis factor-α in lung adenocarcinoma cell line

 Ju-yong Wanga, Ye Jinb, Zhen-ye Xua, Zhan Zhenga   

  1. a Tumor Institute of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
    b Department Bioengineering, School of Life Sciences, Shanghai University, Shanghai 200444, China
  • Received:2008-11-03 Accepted:2008-12-22 Online:2009-03-20 Published:2009-03-15

Objective

To study the effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine, on gene expression of nuclear factor-κB (NF-κB) activated by tumor necrosis factor-α (TNF-α) in lung adenocarcinoma cell line (A549).
Methods

A549 cells were incubated with rat serum containing Feiyanning at different concentrations for 24 and 48 h, respectively. Morphology of cells was observed by an inverted microscope after treatment with reagents. The cell proliferation was examined by 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2, 4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-8) assay. The expressions of NF-κB and inhibitor κBα (IκBα) were studied by Western blotting. NF-κB-dependent luciferase reporter (3×κB-luc) was transfected for 24 h, and the cells were treated with the reagents for 24 h, and then the transcriptional activity of NF-κB promoter was detected by luciferase assay.
Results

TNF-α (1 μg/L) strongly induced the expression of NF-κB by approximately 1.76-fold compared with the control in the nuclei of A549 cells, and the induced NF-κB expression was significantly suppressed by addition of Feiyanning (P<0.01). In addition, Feiyanning inhibited the transcriptional activity of the NF-κB promoter. However, we observed no significant changes in IκBα expression (P>0.05).
Conclusion

Feiyanning Decoction can markedly inhibit human lung cancer A549 cell proliferation, which may be partly due to inhibition of NF-κB activation induced by TNF-α. It is therefore expected to be a new strategy for treating lung cancer.

Key words: Lung neoplasms, Feiyanning Decoction, NF-kappa B, Tumor necrosis factor-a

Figure 1

Effects of serum containing Feiyanning on morphology of A 549 cells (Inverted microscopy, ×400) A: Control serum group; B: 10% serum containing Feiyanning group; C: 15% serum containing Feiyanning group; D: 20% serum containing Feiyanning group."

"

Group n OD value
24 h 48 h Rate of inhibition
after 48 h (%)
Control serum 4 0.325±0.014 0.497±0.021 0
10% serum containing Feiyanning 4 0.337±0.042 0.424±0.018 14.69±1.38
15% serum containing Feiyanning 4 0.344±0.028 0.359±0.020 30.85±1.62*
20% serum containing Feiyanning 4 0.331±0.017 0.323±0.016 36.21±2.01**

Figure 2

Effects of serum containing Feiyanning on NF-κB expression induced by TNF-α in A549 cells Confluent cells were incubated for 48 h with or without 15% serum containing Feiyanning, or TNF-α (1 μg/L). The two panels showed the protein expression levels of NF-κB and TFⅡB; bar graphs showed the expression levels of NF-κB relative to those of TFⅡB. The data were represented asx±s (n=3). Compared with TNF-α, F=14.325, P=0.0054, **P<0.01."

Figure 3

Effects of serum containing Feiyanning on IκBα expression in A549 cells Confluent cells were incubated for 48 h with or without 15% serum containing Feiyanning, or TNF-α (1 μg/L). The two panels showed the protein expression levels of IκBα and α-tubulin; bar graphs showed the expression level of IκBα relative to those of α-tubulin. The data were represented asx±s (n=3)."

Figure 4

Effects of serum containing Feiyanning on NF-κB promoter in A549 cells Cells plated in six-well plates were transfected with 3×κB-luc plasmid. One day after the transfection, the cells were treated with or without 15% serum containing Feiyanning, or TNF-α (1 μg/L) for 24 h, and then luciferase activity in the cell lysates was measured. The experiments were performed in triplicate. The data were represented asx±s (n=3). Compared with TNF-α, F=18.236, P=0.004 67, **P<0.01."

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