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Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (6): 695-700.doi: 10.3736/jcim20120615

Special Issue: Traditional Chinese Medicine

• Original Experimental Research • Previous Articles     Next Articles

Effects of Feiyanning Decoction, a compound traditional Chinese medicine, on iNOS and COX-2 expressions induced by tumor necrosis factor-α in lung adenocarcinoma cell line

Ju-yong Wang(), Jing Guo, Chang-sheng Dong, Zhan Zheng, Qing Wang#br#   

  1. Institute of Traditional Chinese Medicine Oncology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
  • Received:2011-12-09 Accepted:2012-01-12 Online:2012-06-20 Published:2018-06-15
  • Contact: Wang Ju-yong E-mail:wangjuyong1@126.com

Objective: To study the effect of Feiyanning Decoction (FYN), a compound traditional Chinese medicine, on expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) activated by tumor necrosis factor-α (TNF-α) in human lung adenocarcinoma epithelial cell line (A549).
Methods: A549 cells were incubated with rat serum containing FYN for 24 h. Gene expressions of iNOS and COX-2 were determined by quantitative real-time polymerase chain reaction and Western blot. The iNOS-dependent luciferase reporter was transfected for 24 h and the cells were treated with the reagents for 24 h, then the transcriptional activity of iNOS promoter was detected by luciferase assay. The production of NO was determined by diaminofluorescein-2.
Results: FYN significantly inhibited TNF-α-induced expression of iNOS and COX-2 compared with the control group in A549 cells (P<0.01, P<0.01). Also, FYN inhibited the transcriptional activity of the iNOS promoter and reduced NO production compared with the control group (P<0.01, P<0.01).
Conclusion: These results suggest that FYN inhibits iNOS and COX-2 activation induced by TNF-α, therefore, it is expected to develop a new strategy to treat lung cancer.

Key words: lung neoplasms, nitric oxide synthase, cyclooxygenase 2, tumor necrosis factor-alpha, adenocarcinoma, compounds, rats

"

Gene Primer sequence
Forward Reverse Size of PCR
product (bp)
iNOS 5′-ATCCCGAAACGCTACACTT-3′ 5′-AATCCACAACTCGCTCCA-3′ 301
COX-2 5′-CTGTATCCCGCCCTGCTGGTG-3′ 5′-ACTTGCGTTGATGGTGGCTGTCTT-3′ 279
β-actin 5′-AAGTACTCCGTGTGGATCGG-3′ 5′-TCAAGTTGGGGGACAAAAAG-3′ 547

Figure 1

Inhibitory effect of FYN on increase in iNOS mRNA induced by TNF-α in A549 cells The cells were incubated for 24 h. iNOS mRNA was determined by real-time polymerase chain reaction. Data are expressed as mean±standard deviation; n=3; **P<0.01, vs control group; △△P<0.01, vs TNF-α group. a: Control group; b: TNF-α group; c: FYN alone group (15% serum containing FYN); d: FNY plus TNF-α group. FYN: Feiyanning Decoction; iNOS: inducible nitric oxide synthase; TNF-α: tumor necrosis factor-α; A549: human lung adenocarcinoma epithelial cell line."

Figure 2

Inhibitory effect of FYN on increase in iNOS expression induced by TNF-α in A549 cells The cells were incubated for 24 h. iNOS expression was determined by Western blot. Data are expressed as mean±standard deviation; n=3; **P<0.01, vs control group; △△P<0.01, vs TNF-α group. a: Control group; b: TNF-α (1 μg/L) group; c: FYN alone group (15% serum containing FYN); d: FYN plus TNF-α group. FYN: Feiyanning Decoction; iNOS: inducible nitric oxide synthase; TNF-α: tumor necrosis factor-α; A549: human lung adenocarcinoma epithelial cell line."

Figure 3

Inhibitory effect of FYN on increase in NO production induced by TNF-α in A549 cells The cells were incubated for 24 h. The DAF-2 assay was performed. Data are expressed as mean±standard deviation; n=3; **P<0.01 vs control group; △△P<0.01, vs TNF-α group. a: Control group; b: TNF-α group; c: FYN alone group (15% serum containing FYN); d: FYN plus TNF-α group. FYN: Feiyanning Decoction; NO: nitric oxide; TNF-α: tumor necrosis factor-α; A549: human lung adenocarcinoma epithelial cell line."

Figure 4

Effect of FYN on iNOS promoter in A549 cells Cells plated in six-well plates were transfected with iNOS-luciferase plasmid. The cells were treated for 24 h after transfection, and then luciferase activity in the cell lysates was measured. Data are expressed as mean±standard deviation; n=3; **P<0.01, vs control group; △△P<0.01, vs TNF-α group. a: Control group; b: TNF-α group; c: FYN alone group (15% serum containing FYN); d: FYN plus TNF-α group. FYN: Feiyanning Decoction; iNOS: inducible nitric oxide synthase; TNF-α: tumor necrosis factor-α; A549: human lung adenocarcinoma epithelial cell line."

Figure 5

Inhibitory effect of FYN on increase in COX-2 mRNA induced by TNF-α in A549 cells The cells were incubated for 24 h. COX-2 mRNA was determined by real-time polymerase chain reaction. Data are expressed as mean±standard deviation; n=3; **P<0.01, vs control group; △△P<0.01, vs TNF-α group. a: Control group; b: TNF-α group; c: FYN alone group (15% serum containing FYN); d: FYN plus TNF-α group. FYN: Feiyanning Decoction; COX-2: cyclooxygenase-2; TNF-α: tumor necrosis factor-α; A549: human lung adenocarcinoma epithelial cell line."

Figure 6

Inhibitory effect of FYN on COX-2 expression induced by TNF-α in A549 cells The cells were incubated for 24 h. COX-2 expression was determined by Western blotting. Data are expressed as mean±standard deviation; n=3; **P<0.01, vs control group; △△P<0.01, vs TNF-α group. a: Control group; b: TNF-α group; c: FYN alone group (15% serum containing FYN); d: FYN plus TNF-α group; FYN: Feiyanning Decoction; COX-2: cyclooxygenase-2; TNF-α: tumor necrosis factor-α; A549: human lung adenocarcinoma epithelial cell line."

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